liquid chromatography - cosela 1317638748.pdf · 81 4-amino-n10- methylpteroglutamic ... 100 [sar1,...

LIQUID CHROMATOGRAPHY

HPLC Column Compound Index

A1 AAF-Modified Oligonucleotide 9mer

2 AAF-Modified Oligonucleotide

11mer

3 AAF-Modified Oligonucleotide

12mer

4 Acepromazine

5 Acetamide

6 Acetaminophen

7 Acetate

8 Acetic Acid

9 Acetoin

10 Acetone

11 Acetophenazile

12 Acetophenetidine

13 Acetophenone

14 2-Acetoxybenzoic Acid

15 N-Acetyl-2-aminofluorene

Modified 9mer


Modified 11mer


Modified 12mer

18 Acetylcholine

19 Acetylferrocene

20 2-Acetylfuran

21 N-Acetyl-D-Galactosamine

22 Acetyl-Glucosamine

23 N-Acetyl-D-Glucosamine

24 Acetylsalicylic Acid

25 Acid Blue Dye 1

26 Acid Blue Dye 3

27 Acid Blue Dye 9

28 N-Acetylprocainamide

29 cis-Aconitic Acid

30 Acrylamide

31 Acrylic Acid

32 Acrylonitrile

33 Acyclovir

34 Adenine

35 Adenosine

36 Adenosine 5’-diphosphate (ADP)

37 Adenosine 5’-monophosphate

(AMP)

38 Adenosine 5’-monophosphate,

Cyclic (cAMP)

39 Adenosine 5’-triphosphate (ATP)

40 Adipate

41 Adipic Acid

42 Adonitol

43 Adrenaline

44 Alanine

45 Alkanesulfonates C2











56 w-Alkoxy-3,5,7 ...-Polyoxaalkyl

Sulfates (Alkyl Ether Sulfates)

57 1-0-Alkyl-2-Acetyl-sn-Glycero-

3-Phosphocholine (AGEPC)

58 Alkylbenzenesulfonates

59 Alkyl Sulfates

60 Alkyl Sulfates C6






66 Allantoin

67 Allobarbital

68 Allylcyclopentenyl Barbaturic Acid

69 Alphaphenal

70 Alphaprodine

71 Aluminum +3

72 1-Aminoanthraquinone

73 2-Aminoanthraquinone

74 Aminoantipyrine

75 p-Aminoazobenzene

76 p-Aminobenzenesulfonic Acid

77 3-Amino-4-Ethoxyacetanilide

78 4-Amino1-hydroxybutane-1,1-

diphosphonic Acid Monosodium

Salt Trihydrate

79 1-Amino-2-Methylbenzene

80 Aminomethylphosphonic Acid

81 4-Amino-N10-

methylpteroglutamic Acid

82 1-Aminonapthalene

83 Aminooxyisobutyric Acid

84 p-Aminophenol

85 Aminopterin

86 2-Aminopyrimidine

87 Amitriptyline

88 Ammonium

89 Amobarbital

90 AMP

91 cAMP, (Cyclic AMP)

92 AMPA

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Hamilton HPLC Column Compound Index

93 Amphotericin

94 Amphotericin B

95 Amylocaine

96 Androstene-3,17-dione

97 Angiotensin

98 Angiotensin I

99 [Val5] Angiotensin I

100 [Sar1, Thr8] Angiotensin II

101 [Ile7] Angiotensin III

102 [Val4] Angiotensin III

103 Anhydroerythromycin A

104 Anhydro-N-Demethyl-Erythromycin A

105 Anhydro-Tetracycline

106 Aniline

107 o-Anisidine (Methoxyaniline)

108 p-Anisidine

109 Anisole

110 Anthocyanidin

111 Antimicrobials

112 Antimony (III)

113 Antimony (V)

114 Antipyrine

115 Apomyoglobin Tryptic Digest

116 Apooxytetracycline

117 Apramycin

118 Aprobarbital

119 Arabinose

120 Arabitol

121 Arginine

122 Arsenate

123 Arsenic Acid

124 Arsenite

125 Arsenobetaine

126 Arsenocholine

127 Arsenous Acid

128 Ascorbic Acid

129 Asparagine

130 Aspartame

131 Aspartic Acid

132 Aspirin

133 Atropine

134 5-Azacytidine

135 Azelic Acid

136 Azide

137 Aztreonam (E)

138 Aztreonam (Z)

139 Aztreonam Desulfonated

Open-ring Form

140 Aztreonam Ethyl Ester (B)

141 Aztreonam Open-ring Form (E)

142 Aztreonam Open-ring Form (Z)

143 Azure A (Asymmetrical Dimethyl-

Thionin)

144 Azure B

145 Azure C

B146 Barbital

147 Barium

148 Bentazon

149 Benzaldehyde

150 Benzamide

151 Benzene

152 Benzene Pentacarboxylic Acid

153 Benzenesulfonic Acid

154 Benzoate

155 Benzocaine

156 Benzoic Acid

157 Benzonitrile

158 Benzophenonetetracarboxylic Acid

159 3,3’,4,4’-

Benzophenonetetracarboxylic Acid

160 3,3’,4,4’-Benzophenonetetracar

boxylic Acid Dianhydride Ester


boxylic Acid Dimethyl Ester


boxylic Acid Monomethyl Ester


boxylic Acid Tetramethyl Ester


boxylic Acid Trimethyl Ester

165 Benzoylegonine

166 Benzphetamine

167 Benzyl Alcohol

168 Benzylamine

169 Benzylammonium Ion

170 Benzyl Carbamate

171 Benzylcetyldimethylammonium

chloride

172 Benzyl Chloride

173 Benzyldimethylstearylammonium

chloride

174 Benzyldimethyltetradecylammonium

chloride

175 Bethanechol

176 Berberine

177 BHA

178 BHT

179 Biliverdin

180 Biotin

181 Biphenyl

182 Bis(acetylacetonato)cadmium(II)

183 Bis(acetylacetonato)cobalt(II)

184 Bis(acetylacetonato)copper(II)

185 Bis(acetylacetonato)nickel(II)

186 Bisdesethylchloroquine

187 Borate

188 Borontetrafluoride

189 Bovine Cytochrome c

190 Bovine Serum Albumin

191 Bradykinin

192 Bromate

193 Bromite

194 Bromobenzene

195 Brucine

196 Butabarbital

197 Butacaine

198 1,4-Butanediol

199 1,4-Butanedione

200 i-Butanol

201 n-Butanol

202 s-Butanol

203 t-Butanol

204 Butanone

205 Butethal

206 Butirosin A

207 Butirosine

208 Butylamine

209 Butylated Hydroxyanisole

210 Butylated Hydroxytoluene

211 Butyric Acid

212 Butyrophenone

C213 C14-AGEPC

214 C16-AGEPC

215 C18-AGEPC

216 Cadmium

217 Cadmium Phenanthroline

218 Caffeic Acid

219 Caffeine

220 Calciferol

221 Calcium

222 Calmodulin

223 Canine Cytochrome c

224 Cannabinol

225 Capric Acid

226 Caproic Acid

227 Caprylic Acid

228 Carbamazepine

229 Carbonate

230 Carbonic Anhydrase

231 2-Carboxy-D-Arabinitol-1,5-

Bisphosphate

232 2-Carboxy-D-Arabinitol-1-Phosphate

233 2-Carboxy-D-Arabinitol-5-Phosphate

234 2-Carboxy-D-Ribitol-1,5-

Bisphosphate

235 Carvacrole

236 Carvacrol (5-Isopropyl-2-

Methyl-Phenol)

237 Catechin

238 CBZ-l-Azetidinone

239 CBZ-l-Azetidinone-TBA Salt

240 CBZ-l-Serinamide

241 CBZ-l-Serinamide Mesylate

242 CBZ-l-Serine

243 CBZ-l-Threoninamide

244 CBZ-l-Threonine

245 Cesium

246 Chenodeoxycholate, Glycine and

Taurine-conjugated

247 Chenodeoxycholic Acid

248 Chimeric Ribozyme 32mer

249 Chimeric Ribozyme 38mer

250 Chloramphenicol

251 Chloranil (Tetrachloro-1,4-

Benzodinon)

252 Chlorate

253 Chlordiazepoxide

254 Chlordimeform

255 Chlorhexidine

256 Chlorhexidine Gluconate

257 Chlorite

258 Chloroacetic Acid

259 p-Chloroaniline

260 Chlorobenzene

261 p-Chlorobenzenesulfonate

262 3-Chlorobenzohydroxamic Acid

263 3-Chlorobenzoic Acid

264 2-Chloroethylethyl-

Methylsulfonium Ion

265 Chloroform

266 Chlorogenic Acid

267 4-Chloro-2-Methylphenoxy-

Acetic Acid

268 4-(4-Chloro-2-methylphenoxy)-

butanoic Acid

269 2-(4-Chloro-2-methylphenoxy)-

Propionic Acid

270 p-Chloronitrobenzene

271 2-Chlorophenol

272 3-Chlorophenol

273 4-Chlorophenol

274 2-Chlorophenoxyacetic Acid

275 4-Chlorophenoxyacetic Acid

276 2-Chloropropanoic Acid

277 3-Chloropropanoic Acid

278 Chloroquine

279 Chlor-l-succinate

280 Chlor-3-succinate

281 7-Chlorotetracycline

282 8-Chlorotheophylline

283 Chloroxylene

284 Chloroxylenol

285 Chlorphenesin Carbamate

286 Chlorpheniramine

287 Chlorpromazine

288 Chlorpropamine

289 Chlortetracycline

290 4-epi-Chlortetracycline

291 keto-4-Chlortetracycline

292 keto-4-epi-Chlortetracycline

293 Cholate, Glycine and Taurine-

conjugated

294 Cholcalciferol

295 Cholic Acid

296 Choline

297 Chromate

298 Chromium

299 Chromosomal DNA

300 Cinchonidine

301 Cinchonine

302 Ciprofloxacin

303 Cisplatin

304 Citrate

305 Citric Acid

306 Clomesone

307 Clonazepam

308 Clonidine

309 CMP

310 cCMP, (Cyclic CMP)

311 Cobalt

312 Cocaine

313 Codeine

314 Coenzyme Q10

315 Colchicine

316 Conalbumin

317 Concanavalin A

318 Contrathion

319 Copper

320 Copper EDTA

321 Cortisone

322 o-Coumaric Acid

323 p-Coumaric Acid

324 Coumarin

325 Crayfish Carboxypeptidase B

Tryptic Digest

326 Creatinine

327 m-Cresol

328 o-Cresol

329 p-Cresol

330 Cyanate

331 Cyanide

332 Cyanocobalamine

333 Cyclamic Acid

334 Cyclic P3

335 Cyclodisone

336 1,2-Cyclohexanedione

337 1,2-Cyclohexanedione Dioxime

338 Cyclohexanedione Monoxime

339 Cyclohexanone Monoxime

340 Cyproheptadine

341 Cysteine

342 Cytidine

343 Cytidine 5’-diphosphate (CDP)

344 Cytidine 5’-monophosphate (CMP)

345 Cytidine 5’-monophosphate,

Cyclic (cCMP)

346 Cytidine 5’-triphosphate (CTP)

347 Cytochrome

348 Cytosine

D349 2,4-D

350 Daidzein

351 Dansyl-Alanine

352 Dansyl-Arginine

353 Dansyl-Asparagine

354 Dansyl-Glutamine

355 Dansyl-Glycine

356 Dansyl-Leucine

357 Dansyl-Lysine

358 Dansyl-Methionine

359 Dansyl-Phenylalanine

360 Dansyl-Serine

361 Dansyl-Tryptophan

362 Dansyl-Tyrosine

363 Danthron

364 Dapsone

365 DCTA (1,2-Diaminocyclohexane-

Tetraacetic Acid)

366 Decane

367 1-Decanol

368 2-Decanone

369 Decarbamoylgonyautoxin I

370 Decarbamoylgonyautoxin II

371 Decarbamoylgonyautoxin III

372 Decarbamoylgonyautoxin IV

373 Decarbamoylneosaxitoxin

374 Decarbamoylsaxitoxin

375 Dehydroascorbic Acid

376 Dehydroepiandrosterone

377 Dehydroisoascorbic Acid

378 Demeclocycline

379 Demethylchlortetracycline

380 6-Demethylchlortetracycline

381 N-Demethylerythromycin A

382 N-Demethylerythromycin A

Enol Ether

383 Deoxycholate, Glycine and

Taurine-conjugated

384 Deoxycholic Acid

385 2-Deoxyribose

386 Deprotected AAF-Modified

Oligonucleotide 6mer





389 Deprotected Oligonucleotide 4mer



















408 Desethylchloroquine

409 Desethylhydroxychloroquine

410 N-Desmethylmephenytoin

411 N-Desmethylmethsuximide

412 Despropanylfentanyl

413 Desulfonated Aztreonam

414 Detritylated Oligonucleotide 8mer



417 Dexamethasone

418 1,1’-Diacetylferrocene

419 Diallylbarbituric Acid

420 Diallylmethylsulfonium

421 1,2-Diaminocyclohexane-

Tetraacetic Acid

422 cis-Diamminedichloroplatinum (II)

423 Dianisidine (Dimethoxybenzidine)

424 o-Dianisidine

425 Diazepam

426 Dibucane

427 Dibutylamine

428 Dibutylphosphate

429 Dicamba

430 1,2-Dichlorobenzene

431 1,4-Dichlorobenzene

432 2,5-Dichlorobenzene Sulfonic Acid

433 3,3’-Dichlorobenzidine

434 2,4-Dichlorobenzohydroxamic Acid

435 3,6-Dichloro-2-Methoxybenzoic Acid

436 2,3-Dichlorophenol






442 2,4-Dichlorophenoxyacetic Acid

(2,4-D)

443 4-(2,4-Dichlorophenoxy)

butanoic Acid

444 2-(2,4-Dichlorophenoxy)

propionic Acid

445 2,2-Dichloropropanoic Acid

446 2,3-Dichloropropanoic Acid

447 Dicyanodiamide

448 Didrate

449 Diethanolamine

450 Diethylamine

451 Diethylene Glycol

452 Diethylenetriaminepentaacetic Acid

453 Diethylmethylsulfonium

454 Diethylphthalate

455 Diethylstilbestrol

456 Diethyltryptamine

457 Difacinone

458 Difluoro(Acetylacetonato) Boron(III)

459 Diglycolate

460 6,7-Dihydro-7-Hydroxy-1-

Hydroxymethyl-5h-Pyrrolizine

461 Dihydroxyacetone

462 1,2 Dihydroxybenzene

463 2,4-Dihydroxybenzoic Acid

464 Dilaudid

465 Dilevalol

466 Diltiazem

467 Dimethoxybenzidine (Dianisidine)

468 Dimethylallylsulfonium Ion

469 Dimethylamine

470 Dimethylarsinic Acid

471 Dimethylarsinous Acid

472 1-3-Dimethylbarbituric Acid

473 2,5-Dimethylbenzenesulfonic Acid

474 1,1-Dimethylguanidine

475 Dimethyl-2-Hydroxyethyl-Sulfonium

476 2,3-Dimethyl-1-(4-Methyl-Phenyl)-3-

Pyrazolin-5-one

477 Dimethylphthalate

478 Dimethylpropylsulfonium Ion

479 Dimethylselenonium Iodide

480 Dimethylthiourea

481 2,4-Dinitrobenzenesulfonic Acid

482 3,5-Dinitro-o-cresol

483 2,4-Dinitrophenol

484 2,6-Dinitrophenol

485 2,4-Dinitrotoluene-3-sulfonic Acid

486 Dipeptides

487 Diphenoxylate

488 Diphenylguanidine

489 Diphenylhydantoin

490 Diphenyhydramine

491 Dipropyltryptamine

492 Diquat

493 Disodium Edetate

494 2,6-Di-tert-butyl-p-cresol

495 Dithionate

496 Dithiothreitol

497 DMT Protected AAF Modifier


498 DMT Protected Oligonucleotide 4mer




502 DMT Protected Oligonucleotide

10mer


12mer


15mer


16mer


17mer


20mer


21mer


24mer


25mer


28mer


30mer


36mer


44mer


50mer

516 DNA Fragments of p-BR322

517 DNA Oligonucleotide 10mer







524 DNP-Alanine

525 DNP-Arginine

526 DNP-Asparagine

527 DNP-Glutamic Acid

528 DNP-Glutamine

529 DNP-Leucine

530 DNP-Lysine

531 DNP-Methionine

532 DNP-Phenylalanine

533 DNP-Proline

534 DNP-Serine

535 DNP-Threonine

536 DNP-Tryptophan

537 DNP-Tyrosine

538 Dodecane

539 Dodecanesulfonate

540 Dodecyl Sulfate

541 Dog Albumin

542 l-Dopa

543 Dopamine

544 Dowicil-75

545 Doxapram

546 Doxycycline

547 DP2 Oligosaccharide




551 Dyphylline

E552 Edrophonium

553 E D TA

(Ethylenediaminetetraacetic Acid)

554 EDTA, Copper

555 EDTA, Iron

556 Ellagic Acid

557 Enalapril Maleate

558 Endothall Acid

559 Enoxacin

560 Enthanic Acid

561 Ephedrine

562 Epianhydrotetracycline

563 Epicatechin

564 4-Epidoxycycline

565 6-Epidoxycycline

566 4,6-Epidoxycycline

567 Epitetracycline

568 Equine Cytochrome c

569 Ergocristine

570 Ergocristinine

571 a-Ergocryptine

572 a-Ergocryptinine

573 Ergonovine

574 Ergonovinine

575 Ergotamine

576 Ergotaminine

577 Erioglaucine

578 Erythralosamine

579 Erythritol

580 Erythromycin

581 Erythromycin A

582 Erythromycin A Enol Ether

583 Erythromycin B

584 Erythromycin C

585 Erythromycin Related Compound N

586 Estrone

587 Ethanol

588 Ethanolamine

589 Ethonitazene

590 Ethosuximide

591 Ethoxylated Alcohols C6 to C18

592 Ethylamine

593 9-Ethyl-3-Aminocarbazole

594 Ethylbenzene

595 Ethyl Benzoate

596 O-Ethyl S-[2[(diisopropylamino)

ethyl]isopropylphosphonothiolate

597 O-Ethyl S-[2[(diisopropylamino)

ethyl]propylphosphonothiolate

598 O-Ethyl S-[2[(dimethylyamino)

ethyl] -n-propylphosphonothiolate

599 O-Ethyl S-[2[(dimethylyamino)

ethyl] propylphosphonothiolate

600 Ethylene Cyanohydrin

601 Ethylenediamine

602 Ethylenediaminetetraacetic Acid

603 Ethylene Glycol

604 1-Ethylguanidine

605 2-Ethylhexylsalicylate

606 2-Ethylhexyl trans-4-

methoxycinnamate

607 Ethyl Homocholine

608 Ethylmorphine

609 N-Ethylnornicotine

610 5-Ethyl-5-p-Tolylbarbituric Acid

611 Etonitazene

612 Eugenol

613 Everninomycin, SCH 27899

F614 FAD

615 Fenfluramine

616 Fentanyl

617 Fentanyl Homologues and Analogues

618 Ferrocene

619 Ferulic Acid

620 Flavin Adenine Dinucleotide

621 Fluorescein Labeled Oligonucleotide

12mer

622 Fluoride

623 Fluoroacetic Acid

624 Fluoroborate

625 5-Fluorouracil

626 Flurazepam

627 Folic Acid

628 Formaldehyde

629 Formate

630 Formic Acid

631 Fortimicin A

632 Fortimicin B

633 Fructose

634 Fructose-1,6-Diphosphate

635 Fructose-6-Phosphate

636 Fucose

637 Fumaric Acid

638 2-Furaldehyde

639 Furanic Compounds

640 Furfural

641 Furfuryl Alcohol

G642 Galactitol

643 Galactosamine

644 Galactose

645 Galacturonic Acid

646 Gallic Acid

647 Genistein

648 Gentamicin C1

649 Gentamicin C1a

650 Gentamicin C2

651 Gentamicin C2a

652 Gentisic Acid

653 Germall-115

654 Gibberellic Acid

655 Gibberellin A7

656 Glucaric Acid

657 Glucobrassicin

658 Gluconapin

659 Gluconasurtiin

660 Gluconic Acid

661 Glucosamine

662 Glucosamine-1-phosphate

663 Glucosamine-6-phosphate

664 Glucose-6-Phosphate

665 Glucotropaeolin

666 Glucuronic Acid

667 Glufosinate Ammonium

668 Glutamate

669 Glutamic Acid

670 Glutamine

671 g-Glutamylglutamine

672 Glutarate

673 Glutaric Acid

674 Glutathione

675 Glutethimide

676 Glycerine

677 Glycerol

678 Glycine

679 Glycine

conjugated,Chenodeoxycholate

680 Glycine-conjugated Cholate

681 Glycine-conjugated, Deoxycholate

682 Glycine-conjugated, Lithocholate

683 Glycine-conjugated,

Ursodeoxycholate

684 Glycolic Acid

685 Glycylmethoxyacetic Acid

686 Glycylsarcosine

687 Glycyrrhizic Acid

688 Glydant

689 Glyoxal

690 Glyoxylate

691 Glyphosate

692 GMP

693 cGMP, (Cyclic GMP)

694 Gonyautoxin I

695 Gonyautoxin II

696 Gonyautoxin III

697 Gonyautoxin IV

698 Gonyautoxin VIII

699 Guaiacol

700 Guanidine

701 Guanine

702 Guanosine

703 Guanosine 5’-diphosphate (GDP)

704 Guanosine 5’-monophosphate

(GMP)

705 Guanosine 5’-monophosphate,

Cyclic (cGMP)

706 Guanosine 5’-triphosphate (GTP)

H707 HEDTA (Hydroxyethylene-

Diaminetriacetate)

708 Heptane

709 1-Heptanol

710 2-Heptanone

711 Heptanophenone

712 Heptapolyphosphate

713 Hexabarbital

714 Hexamethylenetetramine

715 1-Hexanol

716 2-Hexanone

717 Hexanophenone

718 Hexapolyphosphate

719 Hexylamine

720 Histamine

721 Histidine

722 Hoelan

723 Hyaluronic Acid Decasaccharide

724 Hyaluronic Acid Dodecasaccharide

725 Hyaluronic Acid Hexasaccharide

726 Hyaluronic Acid Octasaccharide

727 Hyaluronic Acid Tetrasaccharide

728 Hydrastine

729 Hydrazine

730 Hydrocortisone

731 Hydrogen Sulfide

732 m-Hydroxybenzaldehyde

733 Hydroxybenzoate

734 2-Hydroxybenzoic Acid (Salicylic Acid)

735 4-Hydroxybenzoic Acid

736 m-Hydroxybenzoic Acid

737 o-Hydroxybenzoic Acid

738 p-Hydroxybenzoic Acid

739 a-Hydroxybutyric Acid

740 g-Hydroxybutyric Acid

741 Hydroxychloroquine

742 9 - {(2-Hydroxyethoxy)

methyl}Guanine

743 2-Hydroxyethylethylmethylsulfonium

744 Hydroxyethyltheophylline

745 4-Hydroxy-Glucobrassicin

746 5-Hydroxyisoquinol

747 7-Hydroxymethotrexate

748 2-Hydroxy-4-methoxybenzophenone

749 5-hydroxymethyl-2-furaldehyde

750 Hydroxymethyl-furfural

751 5-Hydroxy-2-Pyrrolidinone

752 2-Hydroxyquinoline



755 Hyoscyamine

756 Hypophosphite

757 Hypoxanthine

I758 Ibuprofen

759 Imipramine

760 Imminodiacetic Acid

761 Indol-3-Carboxaldehyde

762 Indole-3-Acetic Acid

763 Indomethacin

764 Inosine

765 Inositol

766 Inositol Hexaphosphate

767 Inositol Pentaphosphate

768 Inositol Phosphate

769 Inositol Tetraphosphate

770 Inositol Triphosphate

771 Insulin

772 Iodate

773 Iodide

774 Iron

775 Iron Diethyletriaminepentaacetic

Acid

776 Iron Ethylenediaminetetraacetic

Acid (EDTA)

777 Iron Nitrilotriacetic Acid

778 Iron Phenanthroline

779 Isoascorbic Acid

780 3-Isobutyl-1-Methylxanthine

781 R ,S-2-(4-Isobutylphenyl)-Propionic

Acid

782 Isobutyric Acid

783 Isochlortetracycline

784 Isocitric Acid

785 Isoleucine

786 Isomalt

787 Isomaltose

788 Isomaltotriose

789 Isopropanol

790 3-Isopropyl-(1H)-2,1,3-Benzothiadi-

azin- 4(3H)-one 2,2 Dioxide

791 Isopropyl-6-Methylphenol

792 2-Isopropyl-5-Methylphenol (Thymol)

793 5-Isopropyl-2-Methylphenol (Carvac-

rol)

794 Isopropylmethyl Phosphonic Acid

795 Isoquinoline-N-Oxide

796 Isovaleric Acid

J797 Jacobine

798 Jacozine

K799 Kaempferol

800 Kanamycin

801 Kanamycin A

802 a-Ketobutyric Acid

803 a-ketocaproic Acid

804 a-Ketoglutaric Acid

805 a-Ketoisocaproic Acid

806 a-Ketoisovaleric Acid

807 Ketomalonic

808 6-Keto-Prostaglandin F1a

L809 Labetalol

810 Lactate

811 Lactate Dehydrogenase

812 Lactic Acid

813 Lactose

814 Lauric Acid

815 Lead

816 Leucine

817 Levorphanol

818 LHC II

819 Lidocaine

820 Linamarin

821 Linoleic Acid

822 Linolenic Acid

823 Liothyronine (T3)

824 Lithium

825 Litocholate, Glycine and Taurine-

conjugated

826 Litocholic Acid

827 Lividomycin A

828 Lomefloxacin

829 Lutidine

830 Lymecycline

831 Lysine

832 Lysozyme

833 Lysozyme Tryptic Digest Reduced and

s-Carboxymethylated

M834 Magnesium

835 Malate

836 Maleic Acid

837 Malic Acid

838 d-Malic Acid

839 l-Malic Acid

840 Malonate

841 Malonic Acid,

842 Maltodecaose

843 Maltoheptaose

844 Maltohexaose

845 Maltononaose

846 Maltooctaose

847 Maltopentaose

848 Maltose

849 Maltotetraose

850 Maltotriose

851 Manganese

852 Manganese (III) Protoporphyrin

853 Mannitol

854 Mannose

855 Meclizine

856 Mefenamic Acid

857 Mepenzolate

858 Meperidine

859 Mephensin

860 Mephenytoin

861 Mephobarbital

862 Mepivacine

863 Mercaptoacetic Acid

864 Mescaline

865 Metacycline

866 Metadrenaline

867 Metaphosphate

868 Methacycline

869 Methadone

870 Methagualone

871 Metham Sodium

872 Methanol

873 Methionine

874 Methocarbamol

875 Methotrexate

876 Methoxamine

877 Methoxyacetic Acid

878 Methoxyaniline (o-Anisidine)

879 4-Methoxybenzohydroxamic Acid

880 2-Methoxyethanol

881 2-Methoxyphenol

882 Methsuxamide

883 Methylamine

884 N-Methylaniline

885 Methylarsonic Acid

886 Methylarsonous Acid

887 p-Methylbenzenesulfonic Acid

888 Methyl Benzoate

889 4-Methylbenzohydroxamic Acid

890 B-Methylcholine

891 1-Methyl-2,4-Diaminobenzene

892 Methyl (+)2-(4-(2,4dichlorophe

noxy)) propionate (diclofopmethyl)

893 Methyldopa

894 Methylene Blue

895 4,4’-Methylenedianiline

896 4,4’-Methylenedianiline Bisnadimide

897 4,4’-Methylenedianiline

Mononadamide

898 5-Methyl-2-Furaldehyde

899 Methyl-furfural

900 N-Methylglucamine

901 1-Methylguanidine

902 Methyl Isocyanate

903 Methylmalonic Acid

904 N-Methylmorpholine Oxide

905 N-Methylphenazone

906 Methylphenidate

907 4-Methyl-m-Phenylenediamine

908 3-Methylxanthine

909 Minocycline

910 Molybdate

911 Monobutyl Phosphate

912 Monochloroacetic Acid

913 Monofluorophosphate

914 Monohydrochloride

915 Monomethylarsonic Acid

916 Monosodium Glutamate

917 Morphine

918 Morpholine

919 Muconic Acid

920 Myoglobin Myoglobin Tryptic Digest

921 Myristic Acid

N922 Nadolol

923 Nalidixic Acid

924 Nalorphine

925 Naloxone

926 6-B-Naltrexol

927 Naphthalene

928 2 - [(1-Naphthalenylamino)-

Carbonyl] Benzoic Acid

929 1-Naphthylamine

930 Naproxen

931 Naproxen Sodium

932 Naptalam

933 n-Butanol

934 Neomycin B

935 Neosaxitoxin

936 Neostigmine

937 Niacin

938 Niacinamide

939 Nickel

940 Nicotine

941 Nicotinic Acid

942 Nikethamide

943 Nitrate

944 Nitrilotriacetic Acid

945 Nitrite

946 p-Nitroaniline

947 Nitrobenzene

948 m-Nitrobenzenesulfonate

949 p-Nitrobenzenesulfonic Acid

950 3 - N i t ro - 4 - h y d

roxyphenylarsonic Acid

951 Nitromethane

952 p-Nitrophenol

953 Nonane

954 1-Nonanol

955 2-Nonanone

956 Noradrenaline

957 5-Norbornene-2,3-Dicarboxylic Acid

Anhydride


Dimethyl Ester


Monomethyl Ester

960 Norfloxacin

961 Norleucine

962 Normetadrenaline

963 N-Normorphine

964 NSAID

O965 Octadecylsulfate

966 Octane

967 1-Octanol

968 2-Octanone

969 Ofloxacin

970 Oleandomycin

971 Oleic Acid

972 Oligoribonucleotides

973 Ornithine

974 Orthophosphate

975 Ovalbumin

976 Oxalacetic Acid

977 Oxalate

978 Oxalic Acid

979 Oxaloacetic Acid

980 Oxalosuccinic Acid

981 Oxazepam

982 Oxobis(Acetylacetonato)

Vanadium(IV)

983 Oxolinic Acid

984 Oxytetracycline

P985 Palmitic Acid

986 Panthenol

987 Papaverine

988 Paraquat

989 Paraxanthine

990 Patent Blue VF

991 PCP N-Ethyl Analog

992 PEG 600

993 Pelargonic Acid

994 Pemoline

995 2,3,4,5,6-Pentachlorophenol

996 Pentaerythritol

997 2-Pentanone

998 Pentapolyphosphate

999 Pentazocine

1000 Pentobarbital

1001 Pentylamine

1002 Peptides

1003 Perchlorate

1004 Permanganate

1005 Phenacetate

1006 Phenacetin

1007 Phencyclidine

1008 Phenelzine

1009 Phenobarbital

1010 Phenol

1011 2-Phenylaminonapthalene

1012 4-Phenylbutanoate

1013 Phentermine

1014 Phenylacetate

1015 Phenylalanine

1016 Phenylbutylammonium Ion

1017 10-Phenyldecanohydroxamic Acid

1018 m-Phenylenediamine

1019 o-Phenylenediamine

1020 p-Phenylenediamine

1021 2-Phenylethanol

1022 Phenylethylammonium Ion

1023 Phenylethylmalonamide

1024 Phenylglucuronide

1025 6-Phenylhexanohydroxamic Acid

1026 Phenylmercapturate

1027 9-Phenylnonanohydroxamic Acid

1028 9-Phenylnonanoic Acid

1029 8-Phenyloctanohydroxamic Acid

1030 5-Phenylpentanoate

1031 3-Phenylpropanoate

1032 Phenylpropylammonium Ion

1033 Phenylsulfate

1034 Phorwite RKH

1035 Phosphate

1036 Phosphate Oligomer P1













1049 Phosphatidylcholine

1050 Phosphatidylethanolamine

1051 Phosphite

1052 Phosphoarginine

1053 3-Phosphoglyceric Acid

1054 1-Phosphohistidine

1055 3-Phosphohistidine

1056 N-(Phosphonomethyl)glycine

1057 N-Phosphonomethyliminodiacetic

Acid

1058 Phosphorothioate 28mer

1059 Phosphorothioate deoxycytidine

homopolymer 28mer

1060 Phosphoserine

1061 Phosphothreonine

1062 Phosphotyrosine

1063 Photosystem II Light-Harvesting

Complex (LHC II)

1064 Phthalic Acid

1065 Phylloquinone

1066 Physostigmine

1067 Phytic Acid

1068 Picramic Acid

1069 Pimelic Acid

1070 Pipemidic Acid

1071 Pipenzolate

1072 Piromidic Acid

1073 Plasmid DNA

1074 Polyethylene Glycol

1075 Polyethyene Glycol 600

1076 Polysulfide

1077 Polythiazide

1078 Potassium

1079 Potassium Tetrathionate

1080 Prednisolone

1081 Prednisone

1082 Pregnenolone

1083 Pregnenolone-3-sulfate

1084 Primidone

1085 Procaine

1086 Progesterone

1087 Progoitrin

1088 Proline

1089 L-Proline 3,4-3H(N),histidyl-3-3H(N)

1090 Promazine

1091 Promethazine

1092 Propanoic Acid

1093 1-Propanol

1094 2-Propanol

1095 2-Propanone

1096 Propiomazine

1097 Propionate

1098 Propionic Acid

1099 Propiophenone

1100 Propranolol

1101 Propylamine

1102 Propylene Glycol

1103 Prostacyclin (PGI2)

1104 Protected DNA Oligomers

1105 Proteins

1106 Protocatechuic Acid

1107 Protoporphyrin IX

1108 PTH-Alanine

1109 PTH-Arginine

1110 PTH-Asparagine

1111 PTH-Glutamic Acid

1112 PTH-Glycine

1113 PTH-Leucine

1114 PTH-Lysine

1115 PTH-Phenylalanine

1116 PTH-Tryptophan

1117 PTH-Tyrosine

1118 PTH-Valine

1119 Pyridine

1120 Pyridinoline

1121 Pyridostigmine

1122 Pyridoxine

1123 Pyrilamine

1124 L-Pyroglutamyl- L-histidyl-L-

proline amide

1125 Pyrophosphate

1126 2-Pyrrolidinone

1127 Pyruvate

1128 Pyruvate Dehydrogenase

1129 Pyruvic Acid

Q1130 Quercetin

1131 Quinoline-N-Oxide

1132 Quinolones

R1133 Ranitidine

1134 RB

1135 Reserpine

1136 Resveratrol

1137 Retinol

1138 Retinol Acetate

1139 Retrorsine

1140 Retrorsine N-Oxide

1141 Rhamnose

1142 Ribitol

1143 Riboflavin

1144 Riboflavin 5’-Phosphate

1145 Ribonuclease A

1146 Ribose

1147 Ribose-5-Phosphate

1148 Ribostamycin

1149 Ribozyme 32mer, Chimeric

1150 Ribozyme 38mer, Chimeric

1151 Ribulose-1,5-Diphosphate

1152 RNA

1153 Rolitetracycline

1154 Rubidium

1155 Ruthenium Phenanthroline

S1156 Saccharin

1157 Salicylamide

1158 Salicylic Acid (2-Hydroxybenzoic

Acid)

1159 Saligenin

1160 Saxitoxin

1161 Scopolamine

1162 Scopoletin

1163 Secobarbital

1164 Sedoheptulose-1,7-Diphosphate

1165 Selenate

1166 Selenic Acid

1167 Selenite

1168 Selenocystine

1169 Selenoethionine

1170 Selenohomocystine

1171 Selenomethionine

1172 Selenous Acid

1173 Senecionine

1174 Senecionine N-Oxide

1175 Seneciphylline

1176 Serine

1177 Shikimic Acid

1178 Sialic Acid, (N-Acetyl Neuraminic

Acid)

1179 Silicate

1180 Silver Iodide

1181 Sinalbin

1182 Sinapic Acid

1183 Sinigrin

1184 Sisomicin

1185 SKF 78729A

1186 Slaframine

1187 Sodium

1188 Sorbic Acid

1189 Sorbitol

1190 Soybean Trypsin Inhibitor

1191 Stearic Acid

1192 Streptomycin

1193 Strontium

1194 Strychnine

1195 Suberic Acid

1196 Succinate

1197 Succinic Acid

1198 Succinimide

1199 Succinylsulfathiazole

1200 Sucrose

1201 Sulfacetamide

1202 Sulfadiazine

1203 Sulfadimethoxine

1204 Sulfaguanidine

1205 Sulfamate

1206 Sulfamerazine

1207 Sulfamethazine

1208 Sulfamethizole

1209 Sulfamethoxazole

1210 Sulfanilamide

1211 Sulfanilic Acid

1212 Sulfanilylbenzamide

1213 Sulfapyridine

1214 Sulfate

1215 Sulfathiazole

1216 Sulfide

1217 Sulfisoxazole

1218 Sulfite

1219 Sulfite as

Alphahydroxymethysulfonate

1220 N-Sulfocarbamoyl Toxin B1

1221 N-Sulfocarbamoyl Toxin B2

1222 N-Sulfocarbamoyl Toxin C1




1226 Symmetrical Dimethylthionin

1227 Syringaldehyde

1228 Syringic Acid

T1229 T3

1230 T4

1231 Tartaric Acid

1232 d-Tartaric Acid

1233 l-Tartaric Acid

1234 Tartronic Acid

1235 Taurine

1236 Taurine-conjugated

Chenodeoxycholate

1237 Taurine-conjugated Cholate

1238 Taurine-conjugated Deoxycholate

1239 Taurine-conjugated Lithocholate

1240 Taurine-conjugated

Ursodeoxycholate

1241 99m Technetium

Diethylenetriaminepentaacetic Acid

1242 99m Technetium Glucoheptonate

1243 99m Technetium Methylene

Diphosphonate

1244 99m Technetium Pertechnetate

1245 o-Terphenyl

1246 Testosterone

1247 Tetrabutylammonium Chloride

1248 Tetracaine

1249 2,3,5,6-Tetracarboxybenzene

Sulfonic Acid

1250 Tetrachloro-1,4-Benzodinon

(Chloranil)

1251 2,3,4,5-Tetrachlorophenol



1254 Tetracycline

1255 Tetradecane

1256 Tetradecyl Sulfate

1257 Tetraethylammonium Chloride

1258 Tetramethylammonium Hydroxide

1259 Tetramethylarsonium Iodide

1260 Tetramethylarsonium Ion

1261 Tetramethylenemethylsulfonium Ion

1262 1,1,3,3-Tetramethylguanidine

1263 Tetrapentylammonium Chloride

1264 Tetrapolyphosphate

1265 Tetrapropylammonium Chloride

1266 Tetrathionate

1267 Theaflavin

1268 Theaflavin-3’-digallate

1269 Theaflavin-3-gallate

1270 Theaflavin-3’-gallate

1271 Theafulvins

1272 Thebaine

1273 Theophylline

1274 Thiabendazole

1275 Thiamine

1276 Thiamine Diphosphate

1277 Thiamine Monophosphate

1278 Thiamine Pyrophosphate

Hamilton Company www.hamiltoncompany.com4970 Energy WayReno, Nevada 89502 USAToll Free: 800 648-5950 Telephone: +1-775-858-3000Fax: +1-775-856-7259 Fax: +41-(0)81-660-60-60e-mail: [email protected] e-mail: [email protected] Document No. L80069 © Hamilton Company 10/08 1k

Hamilton Bonaduz AG Via Crusch 8CH-7402 Bonaduz/SwitzerlandToll-Free: 00800-660-660-60Telephone: +41-(0)81-660-60-60Fax: +41-(0)81-660-60-70e-mail: [email protected]

1279 Thiamine Triphosphate

1280 Thiamylal

1281 Thienylcyclohexylpiperidine

1282 Thiobarbituric Acid

1283 Thiochrome

1284 Thiochrome Monophosphate

1285 Thiochrome Pyrophosphate

1286 Thiochrome Triphosphate

1287 Thiocyanate

1288 Thionin

1289 Thiosalicylic Acid

1290 Thiosulfate

1291 Thiourea

1292 Threonine

1293 Thymidine

1294 Thymidine 5’-diphosphate (TDP)

1295 Thymidine 5’-monophosphate (TMP)

1296 Thymidine 5’-triphosphate (TTP)

1297 Thymine

1298 Thymol (2-Isopropyl-5-Methylphenol)

1299 Thymyl Methyl Ether

1300 Thyrotropin-releasing Hormone

1301 Thyrotropin-releasing Hormone

Metabolites

1302 Thyroxine (T4)

1303 Tinopal 5BM-GX

1304 Tinopal CBS-X

1305 Tin Protoporphyrin

1306 Tocopherol

1307 a-Tocopherol

1308 d,l-alpha-Tocopherol

1309 g-Tocopherol

1310 Toluene

1311 o-Toluidine

1312 Transferrin

1313 Trehalose

1314 Triallylsulfonium

1315 Tributylamine

1316 Tricaprin

1317 1,2,3-Trichlorobenzene



1320 2,3,4-Trichlorophenol






1326 2,4,5-Trichlorophenoxyacetic Acid

1327 (+)-2-(2,4,5-Trichlorophenoxy)

propionic Acid

1328 Triclocarban

1329 Tridecane

1330 Triclosan

1331 Triethanolamine

1332 Triethylamine

1333 Trifluoroacetic Acid

1334 1,2,3-Trihydroxybenzene

1335 Trimethylamine

1336 Trimethylarsine Oxide

1337 Trimethylselenonium Iodide

1338 Trimethylselenonium Ion

1339 Trimethylsulfonium Ion

1340 Triphenyltin Hydroxide

1341 Tripolyphosphate

1342 Tris(Acetylacetonato)Aluminum (III)

1343 Tris(Acetylacetonato)Chromium (III)

1344 Tris(Acetylacetonato)Iron (III)

1345 Tris Buffer

1346 Tris(Hexafluoroacetylacetonato)

Chromium (III)

1347 Tris(1-Phenyl-1,3-Butane-Dionato)

Chromiun (III)

1348 Tromethamine

1349 Trypsin

1350 Tryptic Digest of Cytochrome c

1351 Tryptophan

1352 Tyrosine

1353 Tween 80

U1354 Undecane

1355 1-Undecanol

1356 2-Undecanone

1357 Uracil

1358 Urethane

1359 Uric Acid

1360 Uridine

1361 Uridine 5’-Monophosphate (UMP)

1362 Ursodeoxycholate, Glycine and

Taurine-conjugated

1363 Ursodeoxycholic Acid

V1364 Valerate

1365 Valeric Acid

1366 a-Keto, b-Methyl, n-Valeric Acid

1367 Valerophenone

1368 Valine

1369 Vanillic Acid

1370 Vanillin

1371 Verdamicin

1372 Vitamin A

1373 Vitamin B12

1374 Vitamin D2

1375 Vitamin E

1376 Vitamin K1

W1377 Warfarin

1378 Water

X1379 Xanthine

1380 Xanthosine

1381 2,3-Xylenol

1382 2,4-Xylenol

1383 2,5-Xylenol

1384 2,6-Xylenol

1385 3,4-Xylenol

1386 3,5-Xylenol

1387 Xylitol

1388 Xylose

Y1389 Yohimbine

Z1390 Zinc

1391 Zinc Deuteroporphyrin

1392 Zinc Phenanthroline

1393 Zinc Protoporphyrin

HPLC COLUMNS

THEMEASUREOFEXCELLENCETM

HPLC Columns & Bulk Resins

2

The Measure of Excellence

HamiltonFor nearly 50 years, Hamilton Company has been satisfying customer needs in the field of precision fluid measuring. It all started with syringes. Not commercial, mass produced medical syringes, but precision measuring instruments. About 25 years ago Hamilton was the first company to develop and manufacture pressure stable polymeric HPLC columns.

ColumnsWe now offer 19 different polymer-based HPLC columns for reversed phase, anion exchange, cation exchange and ion exclusion separations. Two silica-based C8 and C18 columns are available for reversed phase separations.

Choosing the Right HPLC ColumnBefore you select an HPLC column to separate your sample please check the Hamilton Company HPLC web site at www.hamiltoncompany.com/hplc/app_index_1.asp or e-mail [email protected] for a complete list of compounds that have been separated on Hamilton HPLC columns.

1

Recommended Uses

Reversed Phase PRP®-h1 Long life column for LC/MS applications,synthesized DNA, and small molecule

PRP-h5 Reduced system pressure for protein and peptide separations and enhanced oligonucleotide recovery

PRP-1 General purpose pH stable long life column, synthesized DNA

PRP-3 Gradient protein and peptide separations

PRP-Infinity Nonporous support for very fast gradient separation of large proteins

HxSil C8, C18 General purpose silica-based reversed phase

Anion Exchange PRP-X100 Anions, inorganic and organic using conductivity or UV detection. 0 to 100% solvent compatible.

PRP-X110 Similar to PRP-X100 but for lower level anions (20 ppb to 20 ppm)

PRP-X500 Gradient separation of large proteins and labeled DNA

PRP-X600 Gradient separation of labeled and unlabeled DNA

RCX-10 Isocratic or gradient separation of carbohydrate oligomers up to DP8

RCX-30 Gradient separation of complex carbohydrates

Cation Exchange PRP-X200 Inorganic and organic cations using conductivity or UV detection. Separate mono or divalent cations depending on mobile phase conditions.

PRP-X400 Glyphosate and metabolite in drinking water. Also unique hydrophilic interaction separations

PRP-X800 Mono and divalent cations in the same run. Transition metals

HC-40 Sugar oligomers up to DP8. Max pressure 1,000 psi

HC-75 Mono and disaccharides in corn syrup. Max pressure 1,000 psi Calcium Form

HC-75 Organic acids and sugars. Max pressure 1,000 psi Hydrogen Form

HC-75 Sugar alcohols. Max pressure 1,000 psi Lead Form

Ion Exclusion PRP-X300 Organic acids and alcohols

This brochure only contains a few of the applications which have been performed on Hamilton HPLC columns. In the brochure, columns are grouped by separation mechanism:

reversed phase, anion exchange, cation exchange and ion exclusion.

The Recommended Uses Table below lists a few of the possible uses for each of the column packings.

USP "L" Number

L1 Octadecylsilanechemicallybondedtoporoussilicaorceramic micro-particles,3to10µmindiameter.

L7 Octylsilanechemicallybondedtoporoussilicaparticles,3to10µmindiameter.

L17 Strongcation-exchangeresinconsistingofsulfonatedcross-linked styrene-divinylbenzenecopolymerinthehydrogenform,7to11µmindiameter.

L19 Strongcation-exchangeresinconsistingofsulfonatedcross-linked styrene-divinylbenzenecopolymerinthecalciumform,about9µmindiameter.

L21 Arigid,sphericalstyrene-divinylbenzenecopolymer,5to10µmindiameter.

L23 Ananion-exchangeresinmadeofporouspolymethacrylateorpolyacrylategel withquaternaryammoniumgroups,about10µminsize.

L34 Strongcation-exchangeresinconsistingofsulfonatedcross-linked styrene-divinylbenzenecopolymerintheleadform,about9µmindiameter.

Reversed Phase Support Exchange PoreSupports Material Capacity Size 3 µm 5 µm 7 µm 10 µm 12-20 µm 30-50 µm 50-75µm

PRP-h1 PSDVB* N/A 100Å 79279

PRP-h5 PSDVB* N/A 300Å 79280

PRP-1 PSDVB* N/A 100Å 79578 79579 79580 79581 79582 79583

PRP-3 PSDVB* N/A 300Å 79701 79702

HxSilC8 Silica 719427914379144

HxSilC18 Silica 7913979140 79141

Anion Exchange Support Exchange PoreSupports Material Capacity Size 3 µm 5 µm 7 µm 10 µm 12-20 µm 30-50 µm 50-75µm

PRP-X100 PSDVB*withTrimethyl- 0.19meq/gm 100Å 79584 79585 79586 ammoniunExchanger

PRP-X500 Poly(methacryl- 1.6meq/gm Superfi- 79594 79595 79596 amidoproprylTrimethyl- cially ammoniumchloride porous

PRP-X600 Poly(dimethyl- 1.6meq/gm Superfi- 79597 79598 79599 amidopropyl-methacryl- cially amide) porous

Cation Exchange Support Exchange PoreSupports Material Capacity Size 3 µm 5 µm 7 µm 10 µm 12-20 µm 30-50 µm 50-75µm

PRP-X200 PSDVB*withSulfonate 35µeq/gm 100Å 79587 79588 Exchanger

PRP-X400 PSDVB*withSulfonate 2.5meq/gm N/A 79591 79592 79593 Exchanger

3

HAMILTON Column

HxSil C18

HxSil C8

HC-75 Hydrogen Form

HC-75 Calcium Form

PRP-1PRP-3

PRP-X500

HC-75 Lead Form

USP "L" Number and Hamilton HPLC Columns

USP “L” Numbers & Bulk Resins

*PSDVBisPoly(styrene-divinylbenzene)Bulkresinissoldbythegram.

Bulk Resins

PRP®-1 Reversed Phase HPLC ColumnsReversed Phase HPLC Columns

Benzoic Acid and its Derivatives

PRP®-h1Polymeric 100 Å columns for reversed phase, LC/MS applications.• pH stable from 1 to 13.• Robust (virtually any HPLC solvent can be used).

• One particle size: 5 µm.• Four column diameters: 2.1, 4.6, 10 and 100 mm.• Analytical and semiprep/preparative guard columns.

Naproxen Sodium

0 2 4 6 8 10 Minutes

1

5

3

42

PRP-h1, 5 µ, 100 Å, 4.1 x 50 mm 1. 2,4-Dihdroxybenzoic Acid2. 4-Hydroxybenzoic Acid3. 2-Acetoxybenzoic Acid (Aspirin)4. Benzoic Acid5. 2- Hydroxybenzoic Acid (Salicylic Acid)

Mobil Phase: A: 10 mmol/L sodiumdihydrogenphosphatepH = 2.2 B: Acetonitrile. Gradient: 18% B (Isocratic)Flow Rate:0.60 mL/min. Column Temperature: 50 oCDetection: UV @230 nm. Injection Volumn: 5 µL

0 1 2 3 4 5 Minutes

1 PRP-h1, 5 µm, 100 Å, 4.1 x 50 mm,

1. Naproxen (110 µg/mL)2. Ibuprofen (830 µg/mL)

Mobil Phase: A: 10 mmol/L SodiumdihydrogenphosphatepH = 2 B: Acetonitrile. Gradient: Isocratic 50% A/50%BFlow Rate:0.60 mL/min. Column Tempera ture: 60 oCDetection: UV @230 nm.

2

• Low bleed (ideal for mass spectrometry).

Ordering Information

4

HPLC ColumnsI.D. 50 mm 100 mm 150 mm 250 mm

2.1 mm 79250 79249

4.6 mm 79251 79252 79253 79256

10 mm 79255 79266

100 mm 79523* Analytical guard column for steel columns is 2.3 x 20 mm

Analytical Guard Columns For Steel Columns*Starter Kit 79257(1 holder, 2 cartridges)

Replacement Cartridges (5/pk) 79258

*** Semiprep/preparative guard column for steel columns is 4.6 x 20 mm

Semiprep/Preparative Guard Column For Steel Columns***

Starter Kit 79275(1 holder, 1 cartridge)


5

Polymeric 300 Å columns for oligonucleotide purification and protien separations.• pH stable from 1 to 13.• Robust (virtually any HPLC solvent can be used).• Hydrophilic polymer structure delivers faster oligonucleotide purification.

PRP-h5 Reversed Phase HPLC Columns

• One particle size: 5 µm.• Three column diameters: 2.1, 4.6 and 10 mm.• Analytical and semiprep/preparative guard columns.

4

PRP-h5, 4.1 x 150 mm, 5 µm 300 Å

21-mer Oligonucleotide Purification

0 2 4 6 8 10 Minutes

Mobile phase: A: 0.1 mol/TEAA,ph = 7 B: 0.1 mol/LTEAA, ph = 7, 25% AcetonitrileGradient: 30-80 %B in 15 min. Flow Rate: 0.60 mL/minColumn Temp: 60 oC Detection: UV @ 260 nm

PRP-h5, 5 µ, 300 Å, 4.1 x 50 mm

* Analytical guard column for steel columns is 2.3 x 20 mm

Analytical Guard Columns For Steel Columns*

Starter Kit 79267(1 holder, 2 cartridges)



Starter Kits 79277(1 holder, 1 cartridge)



HPLC ColumnsI.D. 50 mm 100 mm 150 mm 250 mm

2.1 mm 79270 79271

4.6 mm 79261 79262 79272 79273

10 mm 79255 79266


VisittheHAMILTONApplicationCompoundIndexatwww.hamiltoncompany.com/hplc/app_index_1.asp ore-mailchromatography@hamiltoncompany.comforacompletelistofcompoundsthathavebeenseparated.

6

• Five particle sizes: 5, 7, 10, 12-20 and 30-50 µm.• Eleven column internal diameters: 1.0 to 101.6 mm.• Two column materials: 316 stainless steel and PEEK.• Analytical and semiprep/preparative guard columns.

Polymeric 100 Å columns for general reversed phase separations.• pH stable from 1 to 13.• Better sample recovery than silica-based columns (there are no silanol groups).• Excellent durability (stable to any concentration of water or organic solvent).

1. Cytosine2. Uracil3. Uridine

Conditions: 0.05M Citric Acid pH 4.2. Isocratic. Ambient. 1 mL/min. 10 µL, UV 254 nm.

0 30 60 90 Seconds

PRP-1, 4.1 x 50 mm, 3 µm (P/N 79804)

1 2

3

PRP-1 Reversed Phase HPLC Columns

1

2

Small Molecules DNA with Secondary Structure Biocides found in Soap

1. Failure Sequences2. 5’-DMT 36mer

PRP-1, 4.1 x 150 mm, 10 µm (P/N 79425)

Conditions: A) 50mM Sodium Hydroxide pH 12.7; B) 1:1 50mM Sodium Hydroxide : Acetonitrile. Linear Gradient 0% B (0-2 min), 0-50% B (2-25 min). 60°C. 2 mL/min. UV 260nm.

0 10 20

Minutes

1. Chloroxylenol 400 ppm2. Triclocarban 100 ppm

Conditions: 4:1 Acetonitrile : Water. Isocratic. Ambient. 1 mL/min. 10 µL, UV 254 nm.

PRP-1, 4.1 x 250 mm, 7 µm (P/N 79422)

1

2

0 4 8 12 16 Minutes

7


Isomeric C18 Fatty Acids- Chlorhexidine Gluconate Resveratrol

Surfactant Surfactant Fatty Acids

Pyridinoline Diphenylguanidine Folic Acid

0 2 4 6 Minutes

PRP-1, 4.1 x 150 mm, 10 µm (P/N 79425)

1. Chlorhexidine Gluconate 2 µg

Conditions: 1:30:69 Trifluoroacetic Acid : Acetonitrile : Deionized Water. Isocratic. Ambient. 2 mL/min. 1 µL, UV 254 nm.

1

0 3 6 9 12 Minutes

PRP-1, 4.1 x 250 mm, 7 µm (P/N 79422)

1. Resveratrol

Conditions: 3:7 Acetonitrile : 0.1M Sodium Dihydrogen Phosphate pH 11.1. Isocratic. Ambient. 1 mL/min. 10 µL, UV 280 nm.

1

1. Sodium Octadecylsulfate 200 ppm

Conditions: 4.5:5.5 Acetonitrile : Deionized Water. Isocratic. Ambient. 1 mL/min. 10 µL, Conductivity.

PRP-1, 4.1 x 150 mm,10 µm (P/N 79425)

1

0 2 4 Minutes

1. Sodium Dodecylsulfate 200 ppm

Conditions: 3:7 Acetonitrile : Deionized Water. Isocratic. Ambient. 1 mL/min. 10 µL, Conductivity

PRP-1, 4.1 x 150 mm, 10 µm (P/N 79425)1

0 2 4 6 8 Minutes

PRP-1, 4.1 x 100 mm, 3 µm (P/N 79805)

1. Caproic Acid 2. Caprylic Acid 3. Capric Acid 4. Lauric Acid 5. Palmitic Acid 6. Stearic Acid

Conditions: A) 3:1 Acetonitrile : Water; B) Acetonitrile. Linear Gradient 0-100% B in 2 min. Hold for 15 min. Ambient. 0.5 mL/min. 50 µL, UV 254 nm.

1

2

34

5 6

0 5 10 15 Minutes

PRP-1, 4.1 x 250 mm, 7 µm (P/N 79422)

Conditions: 0.1% Trifluoroacetic Acid in 9:1Acetonitrile : Water. Isocratic. Ambient. 0.8 mL/min. 100 µL, UV 210 nm.

1. Linolenic Acid (C18:3) 2. Linoleic Acid (C18:2) 3. Oleic Acid (C18:1) 4. Stearic Acid (C18:0)

1

2 3

4

0 4 8 12 16 Minutes

PRP-1, 4.1 x 250 mm, 7 µm (P/N 79422)

1. Folic Acid

Conditions: 97.5:2.5 20mM Sodium Phosphate pH 6.2 : Acetonitrile. Isocratic Ambient. 1 mL/min. 5 µL, UV 280 nm.

1

0 5 10

2

PRP-1, 4.1 x 150 mm, 10 µm (P/N 79425)

1. Diphenylguanidine 1 mg/mL

Conditions: 2:8 Acetonitrile : Deionized Water with 0.1 % Trifluoroacetic Acid. Isocratic. Ambient. 2 mL/min. 10 µL, UV 280 nm.

1

0 2 4 Minutes

PRP-1, 4.1 x 150 mm, 10 µm (P/N 79425)

1. Pyridinoline, 3 mg/mL

1

0 2 4 Minutes

Conditions: 1:9 Acetonitrile : Deionized Water with 0.1% Heptafluorobutyric Acid. Isocratic. Ambient. 2 mL/min. 3 µL, UV 280 nm.

8

Visit the HAMILTON Application Compound Index at http://www.hamiltoncompany.com/hplc/app_index_1.asp or e-mail [email protected] for a complete list of compounds that have been separated.


* Analytical guard column for steel columns is 2.3 x 20 mm** Analytical guard column for PEEK columns is 3.0 x 8.0 mm

Analytical Guard Columns For Steel Columns* For PEEK Columns** Semiprep/Preparative Guard Column For Steel Columns***

Starter Kits 79447 79317 Starter Kit 79121(1 holder, 2 cartridges) (1 holder, 1 cartridge)

Replacement Cartridges (5/pk) 79445 79318 Replacement Cartridges (2/pk) 79122


HPLC ColumnsI.D. x Length 5 µm 7 µm 10 µm 12-20 µm

1.0 x 50 mm 79751 79755 79759

1.0 x 100 mm 79752 79756 79760

1.0 x 150 mm 79753 79757 79761

1.0 x 250 mm 79754 79758 79762

2.1 x 50 mm 79195 79554

2.1 x 100 mm 79790

2.1 x 150 mm 79366 79480

2.1 x 150 mm* 79796

2.1 x 250 mm 79390 79391

2.3 x 25 mm 79789**

4.1 x 50 mm 79443

4.1 x 100 mm 79479 79565

4.1 x 150 mm 79444 79529 79425 79714

4.1 x 250 mm 79820 79422 79427 79358

4.6 x 50 mm* 79850 79874

4.6 x 100 mm* 79558

4.6 x 150 mm* 79423 79351

I.D. x Length 5 µm 7 µm 10 µm 12-20 µm 30-50 µm

4.6 x 250 mm* 79571 79380 79381 79848

7.0 x 100 mm 79495 79713

7.0 x 300 mm 79919

7.0 x 305 mm 79795 79545 79426

10.0 x 50 mm 79367

10.0 x 100 mm 79355 79499

10.0 x 150 mm 79204 79349

10.0 x 250 mm 79531 79496

21.5 x 100 mm 79791

21.5 x 150 mm 79532 79497

21.5 x 250 mm 79352 79478 79428

30.0 x 300 mm 79718

50.8 x 100 mm 79498

50.8 x 150 mm 79716

50.8 x 250 mm 79567 79493

101.6 x 250 mm 79525

* PEEK hardware** Cartridge Column, Cartridge Holder P/N 32908 must be purchased separately.


PRP-1, 4.1 x 150 mm, 10 µm (P/N 79425)

Conditions: A) 0.1N Perchloric Acid; B) Acetonitrile. Linear Gradient 5-50%B in 10 min. Ambient. 2 mL/min. 10 µL, UV 210 nm.

1. Niacinamide2. Pyridoxine (B6)3. Thiamine (B1)4. Riboflavin (B2)5. Cyanocobalamine (B12)

1

2

3

4 5

0 2 4 6 8 Minutes

Pregnenolone and Pregnenolone-3-sulfate Water Soluble Vitamins Fat Soluble Vitamins

0 5 10 15 20 Minutes

PRP-1, 4.1 x 250 mm, 7 µm, (P/N 79422)

1. Retinol (Vitamin A) 500 ppm2. Calciferol (Vitamin D2) 40 ppm3. alpha-Tocopherol (Vitamin E) 20 ppm4. Phylloquinone (Vitamin K1) 20 ppm

Conditions: 20:70:10 Tetrahydrofuran :Acetonitrile : Water. Isocratic. Ambient. 1.5 mL/min. 100 µL, UV 280 nm.

1

2

3 4

PRP-1, 4.1 x 250 mm, 7 µm (P/N 79422)

1. Pregnenolone-3-sulfate2. Pregnenolone

Conditions: 7:3 Acetonitrile : 25mM Tetrabutylammonium Hydrogensulfate. Isocratic.Ambient. 1 mL/min. 10 µL, UV 210 nm.

12

0 5 10 15 Minutes

9


• Two particle sizes: 10 and 12-20 µm.• Eight column internal diameters: 1.0 to 50.8 mm.• Two column materials: 316 stainless steel and PEEK.• Analytical and semiprep/preparative guard columns.

Proteins at pH 12.2

Polymeric 300 Å columns for protein separations.• pH stable from 1 to 13.

• Better sample recovery than silica-based columns (there are no silanol groups).• Excellent durability (the support can be cleaned with strong acid or base to remove any sample residue).

PRP-3, 4.1 x 150 mm, 10 µm (P/N 79466)

1. Ribonuclease A 1.0 mg/mL2. Insulin 1.0 mg/mL3. Cytochrome c 1.0 mg/mL4. Trypsin 1.0 mg/mL5. Lysozyme 1.0 mg/mL6. Pyruvate Dehydrogenase 0.5 mg/mL

Conditions: A) 0.1% TFA in 50mM Sodium Hydroxide pH 12.2; B) 0.1% TFA in Acetonitrile. Linear Gradient 0-60% B in 30 min. Ambient. 2 mL/min. 50 µL, UV 220 nm.

12

3

45

6

0 10 20 30 Minutes


Analytical Guard Columns For Steel Columns* For PEEK Columns** Semiprep/Preparative Guard Column For Steel Columns*

Starter Kits 79461 79393 Starter Kit 79123(1 holder, 2 cartridges) (1 holder, 1 cartridge)

Replacement Cartridges (5/pk) 79454 79395 Replacement Cartridges (2/pk) 79124

HPLC ColumnsI.D. x Length 10 µm

1.0 x 50 mm 79763

1.0 x 100 mm 79764

1.0 x 150 mm 79765

1.0 x 250 mm 79766

2.1 x 50 mm

2.1 x 100 mm 79863

2.1 x 150 mm 79392

4.1 x 50 mm 79467

4.1 x 100 mm

I.D. x Length 10 µm 12-20 µm

4.1 x 150 mm 79466

4.1 x 250 mm 79794

4.6 x 50 mm* 79539

4.6 x 150mm* 79382

4.6 x 250 mm* 79574

7.0 x 305 mm 79468

10.0 x 250 mm 79526

21.5 x 250 mm 79469

50.8 x 250 mm 79875*PEEKhardware

* Analytical guard column for steel columns is 2.3 x 20 mm * Semiprep/preparative guard column for steel columns is 4.6 x 20 mm** Analytical guard column for PEEK columns is 3.0 x 8.0 mm

0 4 8 Minutes

PRP-3, 4.1 x 150 mm, 10 µm (P/N 79466)

1. Ribonuclease A 2. Cytochrome c 3. Lysozyme 4. Myoglobin 5. Ovalbumin

Conditions: A) 0.1% TFA in Water pH 2.0; B) 0.1% TFA in Acetonitrile. Linear Gradient 25-50% B in 5 min. Hold 3 min. Ambient. 2 mL/min. 100 µL, UV 215 nm.

1

2

34

5

Proteins

10

PRP-Infinity Reversed Phase HPLC Columns

Proteins • One particle size: 4 µm.

• Three column internal diameters: 2.1 to 10.0 mm.

• One column material: 316 stainless steel.

Polymeric nonporous packing for fast gradient separation of large proteins.

• pH stable from 1 to 13.

• Better sample recovery than silica-based columns (there are no silanol groups).• Excellent durability (the support can be cleaned with strong acid or base to remove any sample residue).

PRP-Infinity, 4.1 x 33 mm, 4 µm (P/N 79470)

Conditions: A) 0.1% TFA in Water pH 2.0; B) 0.1% TFA in Acetonitrile. Linear Gradient 21-60% B at 0.6% per second. Ambient. 2 mL/min. 20 µL, UV 215 nm.

1. Ribonuclease A 2. Cytochrome c 3. Transferrin 4. Bovine Serum Albumin 5. Concanavalin A 6. Ovalbumin

1

234

5

6

0 30 60 90 Minutes

ForPRP-Infinitycolumnsanin-linefilterisrecommendedasaguardcolumnisnotavailable.


2.1x33mm 79576

2.1x100mm 79748

4.1x33mm 79470

4.1x50mm 79533

10.0x60mm 79527


Proteins

PRP-Infinity, 4.1 x 33 mm, 4 µm (P/N 79470)

Conditions: A) 0.1% TFA in Water pH 2.0; B) 0.1% TFA in Acetonitrile. Linear Gradient 20-60% B in 10 min. Ambient. 2 mL/min. 20 µL, UV 215 nm.

1. Ribonuclease A 2. Cytochrome c 3. Transferrin 4. Bovine Serum Albumin 5. Concanavalin A 6. Ovalbumin 1

2 5

3

4

6

0 5 10 Minutes


11

HxSil C8 & C18 Reversed Phase HPLC ColumnsSilica-based 100 Å columns for general reversed phase separations.• Separation of closely related PAHs.• Better retention of poorly retained compounds.• Separation of neutral, acidic and basic compounds without tailing (tailing factor of less than 2.0).

• Two particle sizes: 3 and 5 µm.• Five column internal diameters: 2.1 to 21.5 mm.

• Analytical and semiprep/preparative guard columns in 316 stainless steel.

HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

Conditions: 85:15 Methanol : Water. Isocratic. Ambient. 1 mL/min. 5 uL, UV 308 nm.

1. Oxybenzone2. Octocrylene3. Octyl Methoxycinnamate4. Avobenzone (Parsol 1789)5. Octyl Salicylate

1

2

3

45

0 5 10 15 20 Minutes

Meclizine in Tablets Tetracycline by USP Method Oxytetracycline and Tetracycline

0 10 20 Minutes

HxSil C18, 4.6 x 250 mm, 5 µm (P/N 79869)

1. Tetracycline 0.5 mg/mL

Conditions: 680 mL 0.1N Ammonium Oxalate, 270 mL Dimethylformamide, 50 mL 0.2M Dibasic Ammonium Phosphate pH 7.6. Isocratic. Ambient. 1 mL/min. 10 µL, UV 280 nm.

1

0 5 10 Minutes

HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)

Conditions: 680 mL 0.1N Ammonium Oxalate, 270 mL Dimethylformamide, 50 mL 0.2M Dibasic Ammonium Phosphate pH 7.6. Isocratic. Ambient. 1 mL/min. 20 µL, UV 280 nm.

1. Oxytetracycline 0.5 mg/mL2. Tetracycline 0.5 mg/mL

1

2

Sunscreens in Lip Balm Sunscreen Compounds Diphenhydramine in Itch Relief Gel

HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

1. 2-Hydroxy-4- methoxybenzophenone2. 2-Ethylhexyl trans-4- methoxycinnamate3. 2-Ethylhexylsalicylate


2

3

1

0 5 10 Minutes

HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

1. Diphenhydramine 10 ppm

1

0 2 4 Minutes

Conditions: 9:1 Methanol : 20mM Potassium Hydrogen Phosphate pH 7.0. Isocratic. Ambient. 2 mL/min. 5 µL, UV 254 nm.

1. Meclizine 1 mg/mL

Conditions: 9:1 Methanol : 20mM Potassium Hydrogen Phosphate pH 7.0. Isocratic. Ambient. 2 mL/min. 5 µL, UV 254 nm.

1

0 5 10 Minutes

HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

12



HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

1. Uracil 2. Caffeine 3. Phenol 4. Toluene 5. Butylbenzene 6. Triphenylene 7. Amylbenzene 8. Terphenyl

1

2

34 5

6

7

8

0 10 20 Minutes

HxSil C8 & C18 Reversed Phase HPLC Columns

HPLC ColumnsI.D. x Length 3 µm 5 µm

2.1 x 33 mm 79888

2.1 x 50 mm 79889 79881

2.1 x 75 mm 79890 79882

2.1 x 100 mm 79891 79883

2.1 x 150 mm 79892 79884

2.1 x 250 mm 79885

4.6 x 33 mm 79886

I.D. x Length 3 µm 5 µm

4.6 x 50 mm 79872 79867

4.6 x 100 mm 79887 79879

4.6 x 150 mm 79873 79868

4.6 x 250 mm 79869

7.0 x 250 mm 79880

10.0 x 250 mm 79870

21.5 x 250 mm 79871

HxSil C18 Ordering Information

Analytical Guard Column For Steel Columns* Semiprep/Preparative Guard Column For Steel Columns*

Starter Kit 79459 Starter Kit 79137(1 holder, 2 cartridges) (1 holder, 1 cartridge)

Replacement Cartridges (5/pk) 79452 Replacement Cartridges (2/pk) 79138

* Analytical guard column for steel columns is 2.3 x 20 mm * Semiprep/preparative guard column for steel columns is 4.6 x 20 mm

Folic Acid, Methotrexate by USP Method Antibiotics Small Molecules

0 5 10 15 Minutes

HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)

Conditions: 3:1 0.05M Potassium Phosphate pH 7 : Acetonitrile. Isocratic. Ambient. 2 mL/min. 5 µL, UV 254 nm.

1. Ampicillin2. Oxacillin3. Dicloxacillin

1

2 3

0 5 10 Minutes

HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

1. Folic Acid 100 µg/mL2. Methotrexate 100 µg/mL

Conditions: 9:1 0.126M Sodium Phosphate, Dibasic, 0.037M Citric Acid pH 6.0 : Acetonitrile. Isocratic. Ambient. 1.2 mL/min. 10 µL, UV 302 nm.

1

2

HxSil C8 Ordering InformationHPLC ColumnsI.D. x Length 3 µm 5 µm

2.1 x 33 mm 79116

2.1 x 50 mm 79117 79107

2.1 x 75 mm 79118 79108

2.1 x 100 mm 79119 79109

2.1 x 150 mm 79120 79110

2.1 x 250 mm 79111

4.6 x 33 mm 79112

I.D. x Length 3 µm 5 µm

4.6 x 50 mm 79113 79100

4.6 x 100 mm 79114 79101

4.6 x 150 mm 79115 79102

4.6 x 250 mm 79103

7.0 x 250 mm 79104

10.0 x 250 mm 79105

21.5 x 250 mm 79106

* Analytical guard column for steel columns is 2.3 x 20 mm * Semiprep/preparative guard column for steel columns is 4.6 x 20 mm

Analytical Guard Column For Steel Columns* Semiprep/Preparative Guard Column For Steel Columns*Starter Kit 79458 Starter Kit 79135(1 holder, 2 cartridges) (1 holder, 1 cartridge)

Replacement Cartridges (5/pk) 79451 Replacement Cartridges (2/pk) 79136

13

Anion Exchange HPLC Columns

Polymeric anion exchange packings for separation of inorganic and organic anions.

• Easily separate the eight common anions (fluoride through sulfate).• Good separation of fluoride from the water dip.

• Three particle sizes: 5, 10 and 12-20 µm.• Eight column internal diameters: 1.0 to 50.8 mm.

• Two column materials: 316 stainless steel and PEEK.• Analytical and semiprep/preparative guard columns.

PRP-X100

PRP-X100, 4.1 x 50 mm, 3 µm (P/N 79810)

Conditions: 4mM p-Hydroxybenzoic Acid pH 8.5 w/2.5% Methanol 1 mL/min. 100 µL, Conductivity.

1. Fluoride2. Chloride3. Nitrite4. Bromide5. Nitrate

1

2

3

4 5

0 2.5 5 Minutes

• Use with organic solvent from 0 to 100 % for elution of hydrophobic anions or column cleaning.• pH stable from1 to 13.• Use conductivity or UV detection.

Common Anions Anions by Indirect UV Detection EDTA

0 6 12 Minutes

1. Copper EDTA 10 ppm

Conditions: 3mM Sulfuric Acid. Isocratic. Ambient. 2 mL/min. 100 uL, UV 254 nm.

1

PRP-X100, 4.6 x 150 mm, 10 µm (P/N 79354)

0 2 4 6 8 Minutes

1. Fluoride 10 ppm2. Carbonate 10 ppm3. Chloride 10 ppm4. Nitrite 10 ppm5. Bromide 10 ppm6. Nitrate 10 ppm7. Phosphate 10 ppm8. Sulfate 10 ppm

Conditions: 4mM p-Hydroxybenzoic Acid pH 8.9 with 2.5 % Methanol. Isocratic. Ambient. 2 mL/min.100 µL, Indirect UV 310 nm.

1

23

45 6 7 8

PRP-X100, 4.1 x 150 mm, 10 µm (P/N 79434)

Organic Acids Germall 115 Bromate and Other Anions

Sulfite and Sulfate Phosphate and Phosphite Chlorite and Chlorate

14

PRP-X100 Anion Exchange HPLC Columns

PRP-X100, 4.6 x 150 mm, 10 µm (P/N 79354)

1, 2. Germall 115 1 mg/mL

Conditions: 7:3 20mM Sodium Hydroxide : Methanol. Isocratic. Ambient. 2 mL/min. 50 µL, UV 220 nm.

1

2

0 5 10 Minutes

PRP-X100, 4.1 x 150 mm, 10 µn (P/N 79434)

1. Fluoride 20 ppm2. Chloride 20 ppm3. Bromate 40 ppm4. Bromide 20 ppm5. Nitrate 20 ppm

Conditions: 1.7mM Sodium Bicarbonate, 1.8mM Sodium Carbonate, 0.1mM Sodium Thiocyanate. Isocratic. Ambient. 2 mL/min. 10 µL, Suppressed Conductivity.

1

23

4 5

0 2 4 6 8 Minutes

1. Formic Acid 10 µg2. Fluoroacetic Acid 10 µg3. Isopropylmethyl Phosphonic Acid 10 µg4. Chloroacetic Acid 10 µg

Conditions: 0.1M Sodium Hydroxide in 5:95 Acetonitrile : Deionized Water. Isocratic. Ambient. 2 mL/min. 10 µL, Conductivity.

1 2

3 4

0 5 10 Minutes

PRP-X100, 4.1 x 150 mm, 10 µm (P/N 79434)

PRP-X100, 4.1 x 250 mm, 10 µm (P/N 79433)

1. Fluoride2. Chlorite3. Chloride4. Nitrite5. Phosphate6. Bromide7. Nitrate8. Chlorate

Conditions: 4.0mM 4-Amino-2-Hydroxybenzoic Acid pH 5.8. Isocratic. Ambient. 2 mL/min.100 µL, Indirect UV 310 nm.

1

3

2

4

5 6 78

0 2 4 6 8 10 Minutes

PRP-X100, 4.1 x 150 mm, 10 µm (P/N 79434)

Conditions: 20mM Succinic Acid pH 2.9. Isocratic. Ambient. 2 mL/min. 100 µL Conductivity.

1. Fluoride 10 ppm2. Phosphate 50 ppm3. Phosphite 50 ppm4. Chloride 10 ppm

1

2

3

4

0 2 4 6 8 10 12 14 16 Minutes

PRP-X100, 4.6 x 150 mm, 10 µm (P/N 79354)

Conditions: 80:20 8mM p-Hydroxybenzoic Acid pH 10.0 : Methanol. Isocratic. Ambient.2 mL/min. 10 µL, Conductivity.

1. Sulfite 1.27 mg/mL2. Sulfate 0.5 mg/mL

1

2

0 2 4 6 Minutes

****Semiprep/preparativeguardcolumnforsteelcolumnsis4.6x20mm

Analytical Guard Columns For Steel Columns** For PEEK Columns*** Semiprep/Preparative Guard Column For Steel Columns****

StarterKits 79448 79383 StarterKit 79125(1holder,2cartridges) (1holder,1cartridge)

ReplacementCartridges(5/pk) 79446 79385 ReplacementCartridges(2/pk) 79126

**Analyticalguardcolumnforsteelcolumnsis2.3x20mm***AnalyticalguardcolumnforPEEKcolumnsis3.0x8.0mm

HPLC ColumnsI.D. x Length 5 µm 10 µm

1.0x50mm 79767 79771

1.0x100mm 79768 79772

1.0x150mm 79769 79773

1.0x250mm 79770 79774

2.1x50mm 79150 79749

2.1x150mm 79421

2.1x150mm* 79852 79853

2.1x250mm 79190 79346

4.1x50mm 79365

4.1x100mm 79538 79439

4.1x150mm 79434

PEEKhardware


I.D. x Length 5 µm 10 µm 12-20 µm

4.1x250mm 79433 79359

4.6x150mm* 79174 79354

4.6x250mm* 79181 79455

7.0x305mm 79364

10.0x150mm 79715

10.0x250mm 79534

21.5x100mm 79543

21.5x150mm 79542

21.5x250mm 79535 79353

50.8x150mm 79551

15

Anions

PRP-X110 Anion Exchange HPLC ColumnsPolymeric anion exchange packings for separation of inorganic and organic anions from 20 ppb to 20 ppm.• Good separation of fluoride from the water dip.

• Compatible with conductivity or UV detectors.• Use with organic solvent up to 100%• pH stable from 1 to 13.

• One particle size: 7 µm.• Four column internal diameters: 1.0 to 4.6 mm.

• Two column materials: 316 stainless steel and PEEK.• Analytical guard columns.

PRP-X110S, 4.1 x 250 mm, 7 µm (P/N 79735)

1. Fluoride 2 ppm2. Chloride 2 ppm3. Iodide 10 ppm4. Sulfate 10 ppm5. Thiosulfate 10 ppm6. Perchlorate 10 ppm

Conditions: 5mM Sodium Hydroxide with 5mM Phenol, 0.05mM Sodium Thiocyanate. Ambient. 1.5 mL/min. 30 µL, Suppressed Conductivity.

1

2

3

4

5

6

0 7 14 Minutes

Polarizable Anions Glucaric Acid

PRP-X110, 4.1 x 150 mm, 7 µm (P/N 79732)

1. Glucaric Acid 1000 ppm

Conditions: 3:7 Acetonitrile : 20mM Potassium Phosphate, pH 6.2. Isocratic. Ambient. 1.5 mL/min. 20 µL, UV 210 nm.

1

0 6 12 Minutes

* PEEK hardware

HPLC Columns PRP-X110 PRP-X110SI.D. x Length 7 µm 7 µm

1.0 x 50 mm 79775

1.0 x 100 mm 79776

1.0 x 150 mm 79777

1.0 x 250 mm 79778

2.1 x 100 mm* 79742 79743

2.1 x 150 mm* 79744 79745

2.1 x 250 mm* 79746 79747

4.1 x 100 mm 79730 79731

4.1 x 150 mm 79732 79733

4.1 x 250 mm 79734 79735



Analytical Guard Columns For Steel Columns* For PEEK Columns**

Starter Kits 79726 79727(1 holder, 2 cartridges)

Replacement Cartridges (5/pk) 79728 79729

PRP-X110 PRP-X110SI.D. x Length 7 µm 7 µm

4.6 x 100 mm* 79736 79737

4.6 x 150 mm* 79738 79739

4.6 x 250 mm* 79740 79741

1. Fluoride 1 ppm2. Chloride 1 ppm3. Nitrite 1 ppm4. Bromide 1 ppm5. Nitrate 1 ppm6. Phosphate 1 ppm7. Sulfate 1 ppm

0 4 8 Minutes

Conditions: 2.0mM p-Hydroxybenzoic Acid pH 9.3. Isocratic. Ambient. 2 mL/min. 100 µL, Non-Suppressed Conductivity.

1

2

3

4

5

67

PRP-X110, 4.1 x 150 mm, 7 µm (P/N 79732)

16* PEEK hardware

Proteins


• One particle size: 7 µm.• Three column internal diameters: 2.1 to 10.0 mm.• Two column materials: 316 stainless steel and PEEK.• Higher sample capacity than nonporous supports.

Polymeric strong-base anion exchange packing with limited porosity for separation of proteins.• Fast separation of large proteins.• pH stable from 1 to 13.

PRP-X500, 4.6 x 50 mm, 7 µm (P/N 79474)

Conditions: A) 10mM Tris pH 9.0; B) 10mM Tris pH 9.0, 0.5N Sodium Chloride. Linear Gradient 0-50% B in 2.5 min. Hold for 2.5 min. Ambient. 2 mL/min. 30 µL, UV 254 nm.

1. Myoglobin 7 µg2. Conalbumin 7 µg3. Dog Albumin 77 µg

1 2

3

0 2.5 5 Minutes

DMT-on Antisense Phosphorothioate 18mer

PRP-X500, 4.6 x 50 mm, 7 µm (P/N 79474)

1. DMT-on Phosphorothioate 18mer

1

Conditions: A) 85:15 100mM TRIS pH 8.0 : Acetonitrile; B) 85:15 100mM TRIS pH 4.0, 2.5M Lithium Chloride : Acetonitrile. Linear Gradient 0-100% B in 10 min. 2 mL/min. 100 µL of a 1 mg/ml solution, UV 260nm.

0 10 20 Minutes


2.1 x 50 mm* 79864

2.1 x 250 mm 79555

4.6 x 50 mm* 79474

4.6 x 150 mm* 79573

10.0 x 100 mm 79348

Analytical Guard Column For PEEK Columns*

Starter Kit (1 holder, 2 cartridges) 79319



* Analytical guard column for PEEK columns is 3.0 x 8.0 mm

17


0 10 20 30 40 Minutes

PRP-X600, 4.6 x 50 mm, 7 µm (P/N 79360)

Conditions: A) 10mM TRIS, 1mM EDTA pH 9.0; B) 1N NaCl in A, Linear Gradient 60-67.5% B in 15 min. 67.5%-75% B (15-45 min.). Ambient. 1 mL/min. 5 µL, UV 260 nm.

Polymeric weak-base anion exchange packing for gradient separation of proteins and DNA oligomers.• Limited porosity allows separation of large DNA fragments.• pH stable from 1 to 13.

• One particle size: 7 µm.• Column material: PEEK.

• Guard column in PEEK.• Higher sample capacity than nonporous supports.


4.6 x 50 mm* 79360

4.6 x 100 mm* 79187

4.6 x 150 mm* 79188

4.6 x 250 mm* 79189





*PEEKhardware

* Analytical guard column for PEEK columns is 3.0 x 8.0 mm


0 10 20 30 Minutes

PRP-X600, 4.6 x 50 mM, 7 µm (P/N 79360)

Conditions: A) 85:15 100mM TRIS pH 8.0 : Acetonitrile; B) 85:15 100mM TRIS pH 4.0, 2.5M Lithium Chloride : Acetonitrile. Linear Gradient 0-100% B in 10 min. 2 mL/min. UV 260 nm.

1. DMT-on Phosphorothioate 18mer

1

0 7 14 Minutes

PRP-X600, 4.6 x 50 mm, 7 µm (P/N 79360)

1. Myoglobin 1 mg/mL2. Concalbumin 1 mg/mL3. Ovalbumin 1 mg/mL4. Bovine Serum Albumin 1 mg/mL

Conditions: A) 20mM TRIS pH 9.0; B) 20mM TRIS,pH 9.0, 1.0M Sodium Chloride. Linear Gradient 0 to 15%B in 10 min. Hold 2 min. 2 mL/min. UV 220nm.

12

34

Separation of DNA Fragments DMT-on Phosphorothioate 18mer Proteins

18

RCX-10 Anion Exchange HPLC Columns

0 5 10 15 Minutes

RCX-10, 4.1 x 250 mm, 7 µm (P/N 79440)

1. DP22. DP53. DP104. DP15

Conditions: A) 60mM Sodium Hydroxide; B) 60mM Sodium Hydroxide with 500mM Sodium Acetate. Gradient: 0-100%B in 10 min. 2 mL/min. 20 µL, Pulsed Amperometric, Dual Gold ElectrodeE1=500 mV T1=333 msecE2=800 mV T2=166 msecE3=-600 mV T3=499 msec

1

2

3

4

• One particle size: 7 µm.• Three column internal diameters: 2.1 to 4.6 mm.• Two column materials: 316 stainless steel and PEEK.• Guard columns in stainless steel and PEEK.

Polymeric anion exchange packing for isocratic orgradient separation of carbohydrates.• pH stable from 1 to 13.

• Isocratic separation of mono and disaccharides.• Gradient separation of oligomers up to DP8.• Compatible with PAD, conductivity and RI detectors.

Glycols in Water Acetyl Glucosamine Artichoke Tubers

Conditions: 10mN Sodium Hydroxide. Isocratic. Ambient. 1 mL/min. 50 µL, Pulsed Amperometric, Dual Gold Electrode.E1=650 mV T1=166 msecE2=900 mV T2=166 msecE3=-800 mV T3=500 msec

RCX-10, 4.1 x 250 mm, 7 µm (P/N 79440)

1. Ethylene Glycol 100 ppb2. Propylene Glycol 100 ppb

1

2

0 4 8 Minutes

RCX-10, 4.1 x 250 mm, 7 µm (P/N 79440)

Conditions: 10mN Sodium Hydroxide (purged with Helium during the run) Isocratic. Ambient. 1 mL/min. 50 µL, Refractive Index.

1. Galactosamine 100 ppm2. Glucosamine 100 ppm3. Acetyl-Glucosamine 100 ppm

12

3

0 7 14 Minutes

HPLC Columns I.D. x Length 7 µm

2.1 x 250 mm 79399

4.1 x 250 mm 79440

4.6 x 250 mm* 79388




Starter Kits (1 holder, 2 cartridges) 79462 79378


19


RCX-30 Anion Exchange HPLC Columns

0 5 10 15 Minutes

RCX-30, 4.6 x 150 mm, 7µm (P/N 79370)

1. Fucose2. Galactosamine3. Glucosamine4. Galactose5. Glucose6. Mannose

Conditions: 16mN Sodium Hydroxide. Isocratic. Ambient. 2 mL/min. 15 µL, Pulsed Amperometric 5 µAFS Dual Gold ElectrodeE1=600 mV T1=166 msecE2=600 mV T2=166 msecE3=-800 mV T3=249 msec

1

2

3

4 56

Polymeric anion exchange packing for isocratic or gradient separation of complex carbohydrates.• pH stable from 1 to 13.• Isocratic or gradient modes.

• Higher ion exchange capacity support for separation of closely related carbohydrates.• Compatible with PAD and RI detectors.

• One particle size: 7 µm.• Three column internal diameters: 2.1, 4.1 and 4.6 mm.

• Two column materials: Stainless steel and PEEK.• Guard column in PEEK.


2.1x250mm 79170

4.1x250mm 79803

4.6x150mm* 79370

4.6x250mm* 79877


StarterKit(1holder,2cartridges) 79371

ReplacementCartridges(5/pk) 79372

*PEEKhardware Visit the HAMILTON Application Compound Index at http://www.hamiltoncompany.com/hplc/app_index_1.asp or e-mail [email protected] for a complete list of compounds that have been separated.

*AnalyticalguardcolumnforPEEKcolumnsis3.0x8.0mm

Glycoprotein Monosaccharides Corn Syrup Artichoke Tubers

RCX-30, 4.6 x 150 mm (P/N 79370)

Conditions: A) 60mM Sodium Hydroxide; B) 500mM Sodium Acetate in A. Linear Gradient: 0-50% B in 10 min., then 50-100% B (10-20 min.). Ambient. 1 mL/min. 50 µL, Pulsed Amperometric Dual Gold Electrode.E1=350 mV T1=166 msecE2=900 mV T2=166 msecE3=-800 mV T3=500 msec

1. Glucose2. Fructose3. Sucrose4. DP55. DP106. DP15

1

2

3

4

5

6

0 10 20 30 Minutes

RCX-30, 4.6 x 150 mm (P/N 79370)

Conditions: A) 60mM Sodium Hydroxide; B) 500mM Sodium Acetate in A. Linear Gradient 100% A for 4 min., then 0-100% B (4-15 min.) Ambient. 2 mL/min. 50 µL. Pulsed Amperometric Dual Gold Electrode.E1=350 mV T1=166 msecE2=900 mV T2=166 msecE3=-850 mV T3=500 msec

1 34

5

2 6

78

9

0 10 20 Minutes

1. Glucose2. Fructose3. Maltose4. Maltotriose5. Maltotetraose6. Maltopentaose7. Maltohexaose8. Maltoheptaose9. Maltooctaose

20


* Analytical guard column for steel columns is 2.3 x 20 mm** Analytical guard column for PEEK columns is 3.0 x 8.0 m*** Semiprep/preparative guard column for steel columns is 4.6 x 20 mm





Starter Kits (1 holder, 1 cartridge) 79127


Cation Exchange HPLC Columns

• One particle size: 10 µm.• Six column internal diameters: 1.0 to 10.0 mm.

• Two column materials: 316 stainless steel and PEEK.• Analytical and semiprep/preparative guard columns.

PRP-X200Polymeric cation exchange packing for separation of inorganic cations and organic cations.• Easily separate mono or divalent cations.• pH stable from 1 to 13.

• Use with organic solvent up to 100% for elution of hydrophobic cations and column cleaning.• Compatible with conductivity and UV detectors.

PRP-X200, 4.1 x 150 mm (P/N 79441)

1. Lithium2. Sodium3. Ammonium4. Potassium5. Cesium

1

23

4 5

0 5 10 Minutes

Conditions: 2.3:1 4mM Nitric Acid: Methanol. Isocratic. Ambient. 2 mL/min. 100 µL, Conductivity.

Monovalent Cations Glycine and Ethanolamine N-Methylglucamine

PRP-X200, 4.1 x 150 mm (P/N 79441)

1. N-Methylglucamine 1 mg/mL

0 4 8 Minutes

Conditions: 2mN Nitric Acid. Isocratic. Ambient. 2 mL/min. 3 µL, Conductivity.

1

0 6 12 Minutes

Conditions: 2mN Nitric Acid. Ambient. Isocratic. 2 mL/min. 20 µL, Conductivity.

1. Glycine2. Sodium3. Ethanolamine

PRP-X200, 4.1 x 250 mm (P/N 79442)

1

2

3

I.D. x Length 10 µm

1.0 x 50 mm 79779

1.0 x 100 mm 79780

1.0 x 150 mm 79781

1.0 x 250 mm 79782

2.1 x 150 mm 79394

2.1 x 250 mm 79347

4.1 x 100 mm 79363

I.D. x Length 10 µm

4.1 x 150 mm 79441

4.1 x 250 mm 79442

4.6 x 50 mm* 79153

4.6 x 150 mm* 79384

4.6 x 250 mm* 79357

7.0 x 305 mm 79489

10.0 x 250 mm 79549

HPLC Columns

* PEEK hardware* PEEK hardware

21




C18 Guard Column for EPA Glyphosate Application For Steel Columns**



PRP-X400 Semiprep/Preparative Guard Column For Steel Columns***

Starter Kit (1 holder, 1 cartridge) 79131


PRP-X400 Cation Exchange HPLC Columns

*PEEKhardware


2.1 x 250 mm 79398

4.1 x 50 mm 79893

4.1 x 150 mm 79717

4.1 x 250 mm 79473 79563

4.6 x 250 mm* 79387

10.0 x 250 mm 79575

21.5 x 250 mm 79553

Ordering InformationHPLC Columns

* Analytical guard column for PEEK columns is 3.0 x 8.0 m** Analytical guard column for steel columns is 2.3 x 20 mm*** Semiprep/preparative guard column for steel columns is 4.6 x 20 mm

Glyphosate and Metabolite Inositol in Vitamin Matrix Sugar Alcohols

Polymeric cation exchange packing for separation of glyphosate and its metabolite in drinking water.• Faster analysis when compared to competitor’s columns.

• Detection levels of less than 10 ppb when using the post column, OPA derivatization technique.• Applicable to other separations, such as inositol and sugar alcohols.

•Twoparticlesizes:7and12-20µm.•Fivecolumninternaldiameters:2.1to21.5mm.

•Twocolumnmaterials:316stainlesssteelandPEEK.•Analyticalandsemiprep/preparativeguardcolumns.

PRP-X400, 4.1 x 250 mm, 7 µm (P/N 79473)

1. Adonitol 200 ppm2. Arabitol 200 ppm3. Xylitol 200 ppm4. Mannitol 200 ppm5. Sorbitol 200 ppm

Conditions: A) Acetonitrile; B) Water. Linear Gradient 10-30% B in 30 min. Ambient. 2 mL/min. 5 µL, Pulsed Amperometric Dual Gold Electrode; delay 510 msec E1=100mV T1=720msec E2=1000mV T2=120msec E3=-800mV T3=300msec

1

2 4

5

3

PRP-X400, 4.1 x 250 mm, 7 µm (P/N 79473)

Conditions: 85:15 Acetonitrile : 10mN Sodium Hydroxide. Isocratic. Ambient. 2 mL/min. 100 µL, Pulsed Amperometric Dual Gold Electrode.

1, 2, 3, and 5. Unknown4. Inositol 1

2

3

4

5

Conditions: 0.005M Monobasic Potassium Phosphate. Isocratic. Ambient. 0.5 mL/min. 200 µL, Excitation wavelength 338 nm. Emission wavelength 455 nm. Post Column Conditions: Oxidation Solution Flow Rate: 0.20 mL/min; Reaction Coil: 1 mL (0.05 cm ID X 5 m); Reaction Time: 1.4 min; Temp: 38°C Derivatization Solution: 0.30 mL/min; Reaction Coil: 0.20 mL (0.05 cm ID X 1 m), Ambient.

PRP-X400, 4.1 x 250 mm, 7 µm (P/N 79473)

1. Glyphosate2. Aminomethylphosphonic Acid

1

2

Post Column Conditions: 0.1N Sodium Hydroxide at 2 mL/min.

0 5 10 15 20 Minutes 0 10 20 30

Minutes

E1=100 mV T1=716 msecE2=1000 mV T2=166 msecE3=-800 mV T3=299 msecPost Column Conditions: 0.1N Sodium Hydroxide solution at 2 mL/min.

0 6 12 Minutes

22


4.1 x 150 mm 79855

4.1 x 250 mm 79828

4.6 x 150 mm* 79851

4.6 x 250 mm* 79829


* PEEK hardware





Mono and Divalent Cations Transition Metals Transition Metals


PRP-X800 Cation Exchange HPLC Columns

0 4 8 12 16 Minutes

PRP-X800, 4.1 x 250 mm, 7 µm (P/N 79828)

Conditions: 2mM Cupric Sulfate. Isocratic. Ambient. 0.8 mL/min. 10 µL, Indirect UV 220 nm.

1. Lithium 5 ppm2. Sodium 5 ppm3. Ammonium 5 ppm4. Potassium 5 ppm5. Magnesium 5 ppm6. Calcium 5 ppm

1

23

45

6

PRP-X800, 4.1 x 150 mm, 7µm (P/N 79855)

Conditions: A). 2mM Ethylenediamine pH 3.2; B) 100mM Ethylenediamine pH 1.73. Linear Gradient 100% A (0-5 min.), 0-100% B in (5-20 min.) 1 mL/min, 100 µL, Post Column Reaction: 0.0125% 4-(2-pyridylazo) resorcinol (PAR) in 5M Ammonium Hydroxide, 0.5 mL/min. Visible 540 nm.

1. Manganese 6 ppm2. Zinc 6 ppm3. Cadmium 6 ppm4. Lead 6 ppm

1

2

3

4

PRP-X800, 4.1 x 150 mm, 7µm (P/N 79855)

1. Manganese 6 ppm2. Cadmium 6 ppm3. Zinc 6 ppm4. Cobalt 6 ppm

Conditions: A) 10mM Ammonium Acetate. pH 3.4; B) Trifluoroacetic Acid in Water pH 1.5. Linear Gradient 15-35% B in 20 min. 1 mL/min. 100 µL, Post Column Reaction: 0.0125% 4-(2-pyridylazo)resorcinol in 5M Ammonium Hydroxide 0.5 mL/min. Visible 540 nm.

1

2

3 4

0 5 10 15 20 Minutes

0 6 12 Minutes

• One particle size: 7 µm.• Two column internal diameters: 4.1 to 4.6 mm.• Two column materials: 316 stainless steel and PEEK.• Guard columns in 316 stainless steel and PEEK.

Polymeric cation exchange columns for the isocratic separation of mono and divalent cations.• Isocratic separation of lithium, sodium, ammonium, potassium, magnesium and calcium.

• Gradient separation of transition metals.• Excellent durability (stable to any concentration of water or organic solvent).

Gel-Type Cation Exchange Column I.D. x Length 10-15 µm

7.8 x 305 mm 79432

Ordering InformationGuard Columns For Steel Columns*

Hydrogen Form Cation Exchange Starter Kit(1 holder, 1 cartridge) 79133

Hydrogen Form Cation Exchange ReplacementCartridges (2/pk) 79134

Carbonate Form Anion Exchange Starter Kit(1 holder, 1 cartridge) 79866

Carbonate Form Anion Exchange ReplacementCartridges (2/pk) 79865

HC-40 Cation Exchange HPLC Columns

0 8 16 Minutes

HC-40, 7.8 x 305 mm, (P/N 79432)

1. Glucose2. Maltose3. Maltotriose4. Maltotetraose5. Maltopentaose6. Maltohexaose7. Maltooctaose +

Conditions: Water. Isocratic. 80°C. 0.6 mL/min. 12 µL, Refractive Index.

1

2

3

4567

• One particle size: 10 - 15 µm.• One column internal diameter: 7.8 mm.• One column material: 316 stainless steel.

Polymeric 4% cross-linked soft-gel column for cation, ligand exchange separation of carbohydrates.• Separate oligosaccharides up to DP8.• Water mobile phase.• Compatible with RI detectors.

* HC-40 guard column for steel columns is 4.6 x 20 mm

Corn Syrup High Conversion Corn Syrup

0 4 8 12 16 Minutes

HC-40, 7.8 x 305 mm, (P/N 79432)

1. Glucose2. Maltose3. Maltotriose4. Maltotetraose5. Maltopentaose6. Maltohexaose7. Maltoheptaose+

Conditions: Deionized Water. Isocratic. 80°C.0.6 mL/min. 20 µL, Refractive Index.

12

367 45

23

HC-75 Cation Exchange HPLC Columns

• One particle size: 9 µm.• Three column internal diameters: 4.1, 7.8 and 10.0 mm.

• One column material: 316 stainless steel.• Three forms: calcium, lead and hydrogen.

Polymeric 7.5% cross-linked soft-gel columns for cation, ligand exchange separations of carbohydrates. • Compatible with UV and RI detectors.

• Hydrogen form columns for USP L17 applications.• Calcium form columns for USP L19 applications.• Lead form columns for USP L34 applications.

Guard Columns For Steel Columns*

HydrogenFormCationExchangeStarterKit(1holder,1cartridge) 79133

HydrogenFormCationExchangeReplacementCartridges(2/pk) 79134

CarbonateFormAnionExchangeStarterKit(1holder,1cartridge) 79866

CarbonateFormAnionExchangeReplacementCartridges(2/pk) 79865

Ordering InformationGel-Type Cation Exchange Columns 9 µm, 9 µm, 9 µm,I.D. x Length Calcium Lead Hydrogen

4.1 x 250 mm 79431 79476

7.8 x 100 mm 79547

7.8 x 305 mm 79436 79438 79544

10.0 x 250 mm 79528

*HC-75guardcolumnforsteelcolumnsis4.6x20mm


HC-75, 4.1 x 250 mm H+ (P/N 79476)

Conditions: 0.01N Sulfuric Acid. Isocratic. 60°C.0.35 mL/min. 10 µL, UV 210 nm.

1. Citric, Acid 250 ppm2. Lactic Acid 250 ppm3. Acetic Acid 250 ppm

1

2

3

0 6 12 Minutes

Malic Acid Citric, Lactic and Acetic Acids Dihydroxyacetone

1

0 5 10 Minutes

2

3

Conditions: 0.01N Sulfuric Acid. Isocratic. 60°C.0.35 mL/min. 10 µL, UV 210 nm.

1. Maleic Acid 4 ppm2. Malic Acid 1000 ppm3. Fumaric Acid 10 ppm

HC-75, 4.1 x 250 mm H+ (P/N 79476)

24

HC-75, 7.8 x 305 mm Ca+2 (P/N 79436)

1. Formic Acid 0.5 mg/mL2. Glycerol 0.5 mg/mL3. Dihydroxyacetone 0.5 mg/mL

Conditions: 0.01M Calcium Chloride. Isocratic. 90°C. 0.6 mL/min. 5uL, Refractive Index.

1

2

3

0 5 10 Minutes

Ion Exclusion HPLC Columns

0 3 6 Minutes

PRP-X300, 4.1 x 150 mm (P/N 79464)

Conditions: 1mN Sulfuric Acid. Isocratic. Ambient. 1 mL/min. 20 uL, UV 210 nm.

1. Tartaric Acid2. Malic Acid3. Citric Acid4. Lactic Acid5. Acetic Acid

1

2 3

4 5

PRP-X300Polymeric ion exclusion packing for separation of alcohols and organic acids.

• Separate structurally similar alcohols.

• Separate similar organic acids.

• Use with organic solvent from 0 to 100% for elution of highly retained compounds.

• One particle size: 7 µm.

• Five column internal diameters: 1.0 to 10.0 mm.

• Two column materials: 316 stainless steel and PEEK.

• Analytical and semiprep/preparative guard columns.

Fatty Acids Acrylamide and Acrylic Acid Organic Acids

1

2

3

4

56

PRP-X300, 4.1 x 250 mm (P/N 79465)

0 10 20 30 Minutes

1. Acetic Acid 500 ppm2. Propionic Acid 500 ppm3. Isobutyric Acid 500 ppm4. Butyric Acid 500 ppm5. Isovaleric Acid 500 ppm6. Valeric Acid 500 ppm

Conditions: 20:80 t-Butyl Alcohol : 10mM Potassium Phosphate Monobasic pH 2.5. Isocratic. Ambient. 1 mL/min. 100 µL, UV 215 nm.

0 2 4 6 Minutes

PRP-X300, 4.1 x 150 mm (P/N 79464)

Conditions: 95:5 1mN Sulfuric Acid : Acetonitrile. Isocratic. Ambient. 2 mL/min. 5 µL, UV 210 nm.

1. Acrylamide 333 ppm2. Acrylic Acid 350 ppm3. Acrylonitrile 267 ppm

1

2

3





1.0 x 50 mm 79783

1.0 x 100 mm 79784

1.0 x 150 mm 79785

1.0 x 250 mm 79786

2.1 x 150 mm 79396

2.1 x 250 mm 79397

4.1 x 33 mm 79536

4.1 x 50 mm 79356

4.1 x 150 mm 79464

4.1 x 250 mm 79465

4.6 x 150 mm* 79475

4.6 x 250 mm* 79386

10.0 x 250 mm 79572



* Semiprep/preparative guard column for steel columns is 4.6 x 20 mm

Semiprep/Preparative Guard Column For Steel Columns*

Starter Kit (1 holder, 1 cartridge) 79129


Visit the HAMILTON Application Compound Index at http://www.hamiltoncompany.com/hplc/app_index_1.asp or e-mail [email protected]

*PEEKhardware

25

* PEEK hardware23

Lit. No. 80072r2 © Hamilton Company 3/07 5M ACCO Printed in U.S.A.

Sales/Support USA 1-888-525-2123http://www.hamiltoncompany.come-mail: [email protected]

Hamilton Company4970 Energy WayReno, Nevada 89520 USAToll-Free: 800-648-5950Telephone: +1-775-858-3000Fax: +1-775-856-7259e-mail: [email protected]

Hamilton Bonaduz AG Via Crusch 8CH-7402Bonaduz/SwitzerlandTelephone: +41-(0)81-660-60-60Fax: +41-(0)81-660-60-70e-mail: [email protected]

Hamilton Deutschland GmbHDaimlerweg 5A64293 Darmstadt/GermanyTelephone: +49-(0)6151-66706-0Fax: +49-(0)6151-66706-66e-mail: [email protected]

Hamilton Northern EuropeUnit 2, Lyne Riggs EstateLancaster RoadCarnforth, GB-Lancashire LA5 9EA, U.K.Telephone: +44-(0)1524-720-650Fax: +44-(0)1524-720-651e-mail: [email protected]

Hamilton France S.A.R.L.Parc de Haute Technologie-Silic Nº181 Rue Georges BesseF-92182 Antony CedexFranceTelephone: +33-(0)1-55-59-18-18Fax: +33-(0)1-55-59-18-19e-mail: [email protected]

Quality Hamilton Products:MICROLITERTM SyringesGASTIGHT® SyringesChromatography SyringesSyringes for Life ScienceInstrument SyringesSoftGripTM PipettesMiniature ValvesModular Valve PositionerLaboratory Fittings, Adapters & TubingPrecision Syringe PumpsDiluters & DispensersMICROLAB® Robotic WorkstationsElectrochemical SensorsDURACALTM Buffer SolutionsHPLC Columns & ResinsLaboratory Automation for:Drug DiscoveryGenomicsProteomicsForensicsIn Vitro Diagnostics

TRADEMARKS:

PRP® is a registered trademark of the Hamilton Company

THEMEASUREOFEXCELLENCETM

H P L C C O L U M N S

Hamilton Company P.O. Box 10030 Reno, Nevada 89520-0012 Toll-Free 800-648-5950 Tel: +1-775-858-3000 Fax: +1-775-856-7259 e-mail: [email protected] http://www.hamiltoncompany.com

Document No. L80065 © Hamilton Company 2/01 Printed in U.S.A ACCO/10M

9 0 01Call the TOLL FREE ORDER HOTLINE1-888-525-2123 and order from Hamilton with MasterCard, Visa or American Express.

New, HxSilTM C18 and C8 HPLC Columns

Hamilton 5 µm HPLC Columns HxSil C18 HxSil C8 PRP-1 PRP-Infinity5 µm 5 µm 5 µm 4 µm

I.D. x Length Part # Price Part # Price Part # Price Part # Price

2.1 x 30 mm 79576 $330.00

2.1 x 50 mm 79881 $330.00 79107 $330.00

2.1 x 75 mm 79882 $340.00 79108 $340.00

2.1 x 100 mm 79883 $350.00 79109 $350.00 79790 $350.00 79748 $390.00

2.1 x 150 mm 79884 $360.00 79110 $360.00 79366 $390.00

2.1 x 250 mm 79885 $380.00 79111 $380.00

4.6 x 30 mm 79470* $330.00

4.6 x 50 mm 79867 $260.00 79100 $260.00 79443* $315.00 79533* $370.00

4.6 x 100 mm 79879 $360.00 79101 $360.00 79479* $350.00

4.6 x 150 mm 79868 $370.00 79102 $370.00 79444* $390.00

4.6 x 250 mm 79869 $390.00 79103 $390.00 79820* $510.00

7.0 x 250 mm 79880 $650.00 79104 $650.00

10.0 x 250 mm 79870 $995.00 79105 $995.00

21.5 x 250 mm 79871 $1,875.00 79106 $1,875.00* I.D is 4.1 mm

Hamilton 3 um HPLC Columns HxSil C18 HxSil C8 PRP-1 PRP-Infinity 3 µm 3 µm 3 µm 4 µm

I.D. x Length Part # Price Part # Price Part # Price Part # Price

2.1 x 30 mm 79888 $330.00 79116 $330.00 79576 $330.00

2.1 x 50 mm 79889 $330.00 79117 $330.00 79854 $315.00

2.1 x 75 mm 79890 $340.00 79118 $340.00

2.1 x 100 mm 79891 $375.00 79119 $375.00 79844 $350.00 79748 $390.00

2.1 x 150 mm 79892 $380.00 79120 $380.00 79845 $390.00

4.6 x 30 mm 79886 $330.00 79112 $330.00 79470* $330.00

4.6 x 50 mm 79872 $330.00 79113 $330.00 79804* $315.00 79533* $370.00

4.6 x 100 mm 79887 $360.00 79114 $360.00 79805* $350.00

4.6 x 150 mm 79873 $380.00 79115 $380.00 79806* $390.00

* I.D is 4.1 mm

Analytical Guard Columns (for 2.1, 4.1 and 4.6 mm I.D. HPLC Columns)

HxSil C18 HxSil C-8 PRP-1

Starter Kits 79459 79458 79447(1 holder, 2 cartridges) $145.00 $145.00 $145.00

Replacement 79452 79451 79445Cartridges (5/pk) $160.00 $160.00 $160.00

For PRP-Infinity columns an in-line filter is recommended as a guard column is not available.

Semiprep/Preparative Guard Columns (for 7.0, 10.0 and 21.5 mm I.D. HPLC Columns)

HxSil C18 HxSil C8 PRP-1

Starter Kits 79137 79135 79121(1 holder, 1 cartridge) $150.00 $150.00 $150.00

Replacement 79138 79136 79122Cartridges (2/pk) $100.00 $100.00 $100.00


A Word about HPLC ColumnsFor over 20 years Hamilton HPLC columns have

been solving problems no other columns could.

Now we have introduced HxSil silica-based

reversed phase C18 and C8 HPLC columns to

complement our selection of polymer-based

columns. At Hamilton we believe that your

separation requirements dictate the type of

column needed. Whether it’s a silica-based or

polymer-based reversed phase column we are

ready to help. Call us at 800-648-5950 for

expert technical assistance and FREE method

development.

TrademarksThe following company trademarks have been used in this brochureDiscovery® - Sigma-Aldrich Nova-Pak® - WatersInertsil® - GL Sciences PRP®, HxSilTM - Hamilton Company

References1. Robert J. Steffeck, Susan L. Woo, Raymond J. Weigland and James M. Anderson, “A Comparison of Silica-Based C18 and C8 HPLC Columns to Aid Column Selection” LG*GC 13, 9 (1995): 720-726.

Published prices are recommended selling prices as of the date of publication, and are subject to changewithout notice. Prices are in U.S.A. dollars

2

RetentionHamilton HxSil C18 columns exhibit greater retention than most columns. This allows you to separate compounds that are not suffi-ciently retained on other C18 columns. Figures 1 and 2 illustrate theseparation of polycyclic aromatic hydrocarbons (PAH’s) on an HxSilC18 column versus a Nova-Pak® C18 column of the same length. Theflow rate has been reduced for the Nova-Pak C18 column to compen-sate for the smaller I.D. The Hamilton HxSil C18 column providesabout twice the retention for these compounds (23 minute run time

versus 11 minutes). The greater retention of the HxSil C18 column provides better separation of these PAH compounds.With the HxSil C18 column, peaks 2 through 6 (naphthalene, acenaphthylene, acenaphthene, phenanthrene, anthracene) arebaseline resolved. The Nova-Pak C18 column does not fully separate peaks 2 and 3; peaks 5 and 6 are only partiallyresolved. Peaks 9 and 10 are better separated on the HamiltonHxSil C18 column.

Figure 1Polycyclic Aromatic Hydrocarbons (PAH’s)HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

1. Benzene 50 ppm2. Naphthalene 50 ppm3. Acenaphthylene 113 ppm4. Acenaphthene 38 ppm5. Phenanthrene 50 ppm6. Anthracene 50 ppm7. Pyrene 50 ppm8. Chrysene 50 ppm9. Benzo[b]fluoranthene 50 ppm10. Benzo[k]fluoranthene 50 ppm11. 1,2,5,6-Dibenzanthracene 50 ppm

Conditions: 7:3 Acetonitrile : Water.2 mL/min, 50 µL, Isocratic. Ambient. UV at 254 nm.

6

9

11

10

0 10 20 30Minutes

873

2

1

5

Figure 2Polycyclic Aromatic Hydrocarbons (PAH’s)Nova-Pak C18, 3.9 x 150 mm, 5 µm


Conditions: 7:3 Acetonitrile : Water.1.4 mL/min, 50 µL, Isocratic. Ambient. UV at 254 nm.

0 2 4 6 8 10 12 14 16Minutes

11

10

9

3

2

1

8

7

6

4

5

Figure 3Eight Small Molecules HxSil C18, 4.6 x 150 mm,5 µm (P/N 79868)

Figure 4Eight Small Molecules Inertsil ODS-3, 4.6 x 150 mm, 5 µm

1. Uracil 10 ppm2. Caffeine 10 ppm3. Phenol 10 ppm4. Toluene 10 ppm5. n-Butylbenzene 100 ppm6. Triphenylene 10 ppm7. Amylbenzene 100 ppm8. o-Terphenyl 10 ppm

Conditions: 7:3 Acetonitrile :Water. 1 mL/min, 50 µL, Isocratic. Ambient. UV at 254 nm.



SelectivityEach manufacturer’s reversed phase column is different. Choosing the best column to separate your sample can be difficult. Hamiltonmanufactures both silica-based and polymeric reversed phase HPLCcolumns, providing you with a wide range of column retention selectivities and performance benefits. The applications which follow,demonstrate the performance characteristics of Hamilton silica-basedreversed phase columns.

Small MoleculesFigures 3 and 4 compare Hamilton HxSil C18 and Inertsil® ODS-3 columns of the same length and particle size. The HxSil C18 columnprovides slightly longer retention (1 minute), and better separation ofpeaks 7 and 8 (amylbenzene and o- terphenyl). On the Inertsil ODS-3column these two compounds coelute.

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The HPLC Performance Report for each lot of HxSil C18 and C8 support includes information for the parameters listed below.

Silica Lot NumberParticle Size dv 50Particle Distribution dv 90/10Pore VolumeSilica Specific Surface AreaMetals

SodiumIronAluminum

Hamilton Company’s new HxSil silica-based reversed phase C18 and C8 HPLC columns provide greater retention of compoundsthan other C18 and C8 columns. This allows you to separatecompounds that are not adequately retained on other reversedphase columns. When you combine HxSil columns with our selection of polymer-based reversed phase columns you have awide range of column retention selectivity, and performance characteristics from which to choose.

At Hamilton we believe that your separation requirements dictate the type of column needed. Whether it’s a silica-based or polymer-based reversed phase column we are ready to help. Call us at 800-648-5950 for expert technical assistance and FREE method development. We understand that sometimes the only way to know if an HPLC column will separate your compound is to see your sample run on that column.

Each batch of HxSil C18 and C8 support is tested to determineretention and tailing of six model compounds. This mixturedeveloped by Steffeck et al.1 is used to determine the suitability of the C18 bonded phase and behavior of the underlying silicatoward neutral, hydrophobic, acidic and basic compounds. Thetailing factor for each compound is measured at 5% of peakheight (more demanding than measurment at 10% of peakheight).

The purpose of each probe is as follows: Uracil is a void volume marker.Pyridine is a basic compound used to test the activity of silanols toward bases.Phenol is an acidic compound used along with pyridine to determine the underlying silica activity.N,N-Dimethylaniline is a second basic compound used to determine the activity of residual silanols toward bases.4-Butylbenzoic Acid is a second acidic compound used to test the activity of residual silanols toward acids.Toluene tests the hydrophobicity of the column.

Both the Hamilton C18 (Figure 20) and C8 (Figure 21) columns separate all six test probes without a tailing peak (a tailing peak isdefined as a peak with a tailing factor greater than 2.0). This demon-strates the suitability of Hamilton HxSil C18 and C8 columns for theanalysis of acids, bases and neutral compounds. Of the 86 columnstested in the Steffeck et al. paper, 53 (62%) of the columns had 1 ormore of the test probes listed as a tailing peak.

Test Probes

Bonded Phase Lot NumberLigandCarbon Loading %µmol Ligand/m2

Capping Agent

Figure 20Six Probe Test Mix HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


1. Uracil2. Pyridine3. Phenol4. N,N-Dimethylaniline5. 4-Butylbenzoic Acid6. Toluene

Conditions: 65:35 Acetonitrile : 0.05M Sodium Citrate pH 3.15. 1 mL /min, 10 µL, Isocratic. Ambient. UV at 254 nm.



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PeptidesFigures 17 through 19 illustrate the separation of a five componentdecapeptide standard on three Hamilton reversed phase columns.

The peptide standard contains the following five decapeptides;1. NH2-Arg-Gly-Ala-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide2. Acetyl-Arg-Gly-Gly-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide3. Acetyl-Arg-Gly-Ala-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide4. Acetyl-Arg-Gly-Val-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide5. Acetyl-Arg-Gly-Val-Val-Gly-Leu-Gly-Leu-Gly-Lys-amide

Figure 17Five Peptide Standards PRP-1, 4.1 x 150 mm, 5 µm (P/N 79444)

Figure 19Five Peptide Standards HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)

Conditions: A) 0.1%Trifluoroacetic Acid; B) 50% Acetonitrile, 0.09%Trifluoracetic Acid, Linear Gradient 0 -100% B in 50 min. Ambient. 1 mL /min, 100 µL, UV at 215 nm.


Conditions: A) 0.1% Trifluoroacetic Acid; B) 50% Acetonitrile, 0.09% Trifluoracetic Acid, Linear Gradient 0 -100% Bin 50 min. Ambient. 1 mL /min, 100 µL, UV at 215 nm.

Conditions: A) 0.1%Trifluoroacetic Acid; B) 50% Acetonitrile, 0.09% Trifluoracetic Acid, Linear Gradient 0 -100% B in 50 min. Ambient.1 mL /min, 100 µL, UV at 215 nm.

It is useful to note the unique selectivity of the PRP-1 polymeric,poly(styrene-divinylbenzene) HPLC column. It completely separatesall five peptide standards. PRP-1 polymeric columns have the addedadvantage of pH stability from 0 to 14, and they are pressure stable to 5,000 psi. The HxSil C18 and C8 columns separate three of thestandards and only partially resolve the first two peptide standards.None of the C18 silica columns tried (Discovery®, Vydac®, and Nova-Pak®) separated all five peptide standards.

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Technical DetailsHamilton HxSil C18 columns are functionalized mono-merically with octadecyldimethylchlorosilane and endcapped with trimethylchlorosilane. C8 columns arefunctionalized with octyldimethylchlorosilane and endcapped with trimethylchlorosilane.

EfficiencyThe tangent method is used to determine column efficiency. This method is common to a variety of column manufacturers and is easily performed by theuser. Typical efficiencies are on the order of >10,000plates per column for a 150 mm long column.

Reproducibility and SymmetryHxSil C18 and C8 columns are manufactured to provide reproducible retention of your compound. Tight manu-facturing controls and extensive characterization of thebase silica enable us to manufacture C18 columns withreproducible retention. Just look at the results of fourbatches of HxSil C18.

% Carbon 16.6 17.6 17.9 17.9µmol ligand/m2 3.0 3.1 3.2 3.2Particle Size 5 µm 5 µm 5 µm 3 µm

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USP MethodsThe Hamilton HxSil C18 column meets the requirements of a USP L1 column and the HxSil C8 column meets the requirements foran L7 column. Figure 5 illustrates the analysis of tetracycline.

Figure 5Tetracycline by USP Method HxSil C18, 4.6 x 250 mm, 5 µm (P/N 79869)

Figure 6 Folic Acid and Methotrexateby USP Method HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

Figure 7Oxytetracycline andTetracycline HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)


Conditions: 680 mL 0.1NAmmonium Oxalate,270 mL Dimethylformamide, 50 mL 0.2 M DibasicAmmonium Phosphate pH 7.6. 1 mL/min, 10 µL, Isocratic. Ambient. UV at 280 nm.


Conditions: 9:1 0.126 MSodium Phosphate, dibasic,0.037 M Citric Acid pH 6.0 :Acetonitrile. 1.2 mL/min, 10 µL, Isocratic. Ambient. UV at 302 nm.


Conditions: 680 mL 0.1N AmmoniumOxalate, 270 mL Dimethylformamide, 50 mL 0.2 M Dibasic Ammonium Phosphate pH 7.6. 1 mL/min, 20 µL, Isocratic. Ambient. UV at 280 nm.

Figure 8Anticonvulsants HxSil C18, 4.6 x 150 mm,5 µm (P/N 79868)

1. Ethotoin 2 µg 2. Phenobarbital 2 µg3. Methsuximide 2 µg4. Diazepam 0.5 µg

Conditions: A) Water; B) Acetonitrile. Linear Gradient 20-60% B in 10 min. Hold at 60% B for 5 min. Ambient. 1 mL/min, 100 µL, UV at 254 nm.

Figure 6 demonstrates the separation of folic acid from methotrexateon a HxSil C18 “L1” column following the USP 24 recommended conditions.

AnticonvulsantsThree anticonvulsants and one antianxiety drug are separated in a 12 minute gradient run with only an acetonitrile, water mobilephase (see Figure 8). All four peaks have excellent symmetry,narrow band widths and good resolution.

AntibioticsThe separation of oxytetracycline and tetracycline using an HxSil C8 column is shown in Figure 7.

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Figure 13 Seven Proteins HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

Figure 14Seven Proteins HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)

Figure 15 Seven Proteins PRP-1, 4.1 x 150 mm, 5 µm (P/N 79444)

ProteinsHamilton Company o�ers four reversed phase column packings for the separation of proteins: two silica-based packings (HxSil C18 andHxSil C8) and two polymer-based packings (PR P ® -1 and PRP-In�nity).In Figures 13 through 15 seven proteins are separated using the sameconditions. In Figure 16 the protein mixture is a little di�erent andthe �ow rate is optimized (increased) for the nonporous PRP-In�nitycolumn.

1. Ribonuclease A 1mg/mL2. Cytochrome c 1 mg/mL3. Lysozyme 1 mg/mL4. Transferrin 1 mg/mL5. Bovine Serum Albumin

1 mg/mL6. Myoglobin 1 mg/mL7. Ovalbumin 1 mg/mL

Conditions: A) 0.1%Tri�uoroacetic Acid in Water; B) 0.1% Tri�uoroacetic Acid inAcetonitrile. Linear Gradient 20-80% B in 33.3 min. Ambient. 1 mL/min, 30 µL, UV at 215 nm.

Conditions: A) 0.1%Tri�uoroacetic Acid in Water; B) 0.1% Tri�uoroacetic Acid in Acetonitrile. Linear Gradient 20-80% B in 33.3 min. Ambient. 1 mL/min, 30 µL,UV at 215 nm.



1. Ribonuclease A 1 mg/mL2. Cytochrome c 1 mg/mL3. Lysozyme 1 mg/mL4. Transferrin 1 mg/mL5. Bovine Serum Albumin


Conditions: A) 0.1%Tri�uoroacetic Acid in Water; B)0.1% Tri�uoroacetic Acid inAcetonitrile.Linear Gradient 20-80% B in 33.3 min. Ambient. 1 mL/min, 30 µL, UV at 215 nm.

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SteroidsFour steroids are separated on a Hamilton HxSil C18 (Figure 11) andC8 (Figure 12) column of the same length. Excellent selectivity isdemonstrated by both columns (baseline separation of all foursteroids). Two of the steroids, 4-androstene-3,17-dione (peak 2) and

Figure 11SteroidsHxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

1. Hydrocortisone 250 ppm2. 4-Androstene-3,17-dione 250 ppm3. Testosterone 250 ppm4. Progesterone 250 ppm

Conditions: 8:2 Methanol : Water .1 mL/min, 20 uL, Isocratic. Ambient. UV at 240 nm.

Figure 12SteroidsHxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)


Conditions: 8:2 Methanol : Water .1 mL/min, 20 µL, Isocratic. Ambient. UV at 240 nm.

Meclizine in Motion Sickness PillsIn Figure 9 the antiemetic, meclizine is separated from excipients in motion sickness pills. This amine base drug exhibits excellent sym -metry (tailing factor of 1.061) on a Hamilton HxSil C18 column.

Diphenhydramine in Itch Relief GelIn Figure 10 the antihistamine diphenhydramine is separated in just over 2 minutes. Easy sample preparation is characterizedby water dilution, and �ltration. As in Figure 9, the compound ofinterest (diphenhydramine) is an amine base.

Figure 9Meclizine in Motion Sickness PillsHxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


Conditions: 9:1 Methanol :20 mM PotassiumHydrogen Phosphate pH 7.0. 2 mL/min, 5 µL, Isocratic. Ambient. UV at 254 nm.

Figure 10Diphenhydramine in Itch Relief GelHxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


Conditions: 9:1 Methanol : 20 mM Potassium HydrogenPhosphate pH 7.0.2 mL/min, 5 µL, Isocratic. Ambient. UV at 254 nm.

testosterone (peak 3) di�er only by the oxidation state at the C-17position; a carbonyl and hydroxyl group respectively. Note thereduced retention with the C8 column. 4.5 minute run time versus7.5 with the C18.

Figure 16Six Proteins PRP-In�nity, 4.1 x 30 mm, 4 µm (P/N 79470)

1. Ribonuclease A 1mg/mL2. Cytochrome c 1 mg/mL3. Transferrin 1 mg/mL4. Bovine Serum Albumin

1 mg/mL5. Concanavalin A

1 mg/mL6. Ovalbumin 1 mg/mL

Conditions: A) 0.1%Tri�uoroacetic Acid in Water; B) 0.1% Tri�uoroacetic Acid in Acetonitrile. Linear Gradient 20-60% B in 1.2 min. Ambient. 2 mL/min, 20 µL,UV at 215 nm.

The HxSil silica-based C18 and C8 columns show better resolution than the polymer-based PRP-1 and -In�nity columnsfor protein separations. The polymeric PRP-1 and -In�nitycolumns are used for protein puri�cation when columncleaning/chemical sterilization is needed. Cleaning with acid orbase does not harm these polymeric columns. The PRP-In�nitycolumn (Figure 16) is well suited for the fast (less than 2 minute)separation of proteins.

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Figure 13 Seven Proteins HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

Figure 14Seven Proteins HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)

Figure 15 Seven Proteins PRP-1, 4.1 x 150 mm, 5 µm (P/N 79444)

ProteinsHamilton Company offers four reversed phase column packings for the separation of proteins: two silica-based packings (HxSil C18 andHxSil C8) and two polymer-based packings (PRP®-1 and PRP-Infinity).In Figures 13 through 15 seven proteins are separated using the sameconditions. In Figure 16 the protein mixture is a little different andthe flow rate is optimized (increased) for the nonporous PRP-Infinitycolumn.



Conditions: A) 0.1%Trifluoroacetic Acid in Water; B) 0.1% Trifluoroacetic Acid inAcetonitrile. Linear Gradient 20-80% B in 33.3 min. Ambient. 1 mL/min, 30 µL, UV at 215 nm.

Conditions: A) 0.1%Trifluoroacetic Acid in Water; B) 0.1% Trifluoroacetic Acid in Acetonitrile. Linear Gradient 20-80% B in 33.3 min. Ambient. 1 mL/min, 30 µL,UV at 215 nm.



1. Ribonuclease A 1 mg/mL2. Cytochrome c 1 mg/mL3. Lysozyme 1 mg/mL4. Transferrin 1 mg/mL5. Bovine Serum Albumin


Conditions: A) 0.1%Trifluoroacetic Acid in Water; B)0.1% Trifluoroacetic Acid inAcetonitrile. Linear Gradient 20-80% B in 33.3 min. Ambient. 1 mL/min, 30 µL, UV at 215 nm.

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SteroidsFour steroids are separated on a Hamilton HxSil C18 (Figure 11) andC8 (Figure 12) column of the same length. Excellent selectivity isdemonstrated by both columns (baseline separation of all foursteroids). Two of the steroids, 4-androstene-3,17-dione (peak 2) and

Figure 11Steroids HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


Conditions: 8:2 Methanol : Water.1 mL/min, 20 uL, Isocratic. Ambient. UV at 240 nm.

Figure 12Steroids HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)


Conditions: 8:2 Methanol : Water.1 mL/min, 20 µL, Isocratic. Ambient. UV at 240 nm.

Additional Pharmaceutical SeparationsIf the chromatograms shown below don’t convince you Hamilton Company has an HPLC column for your sample, call us at 800-648-5950 for expert technical assistance and FREE method development. We understand that sometimes the only way to know if an HPLC column will separate your compound is to see your sample run on that column.

Meclizine in Motion Sickness PillsIn Figure 9 the antiemetic, meclizine is separated from excipients in motion sickness pills. This amine base drug exhibits excellent sym-metry (tailing factor of 1.061) on a Hamilton HxSil C18 column.

Diphenhydramine in Itch Relief GelIn Figure 10 the antihistamine diphenhydramine is separated in just over 2 minutes. Easy sample preparation is characterizedby water dilution, and filtration. As in Figure 9, the compound ofinterest (diphenhydramine) is an amine base.

Figure 9Meclizine in Motion Sickness PillsHxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


Conditions: 9:1 Methanol :20 mM PotassiumHydrogen Phosphate pH 7.0. 2 mL/min, 5 µL, Isocratic. Ambient. UV at 254 nm.

Figure 10Diphenhydramine in Itch Relief GelHxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


Conditions: 9:1 Methanol : 20 mM Potassium HydrogenPhosphate pH 7.0. 2 mL/min, 5 µL, Isocratic. Ambient. UV at 254 nm.

testosterone (peak 3) differ only by the oxidation state at the C-17position; a carbonyl and hydroxyl group respectively. Note thereduced retention with the C8 column. 4.5 minute run time versus7.5 with the C18. Figure 16

Six Proteins PRP-Infinity, 4.1 x 30 mm, 4 µm (P/N 79470)

1. Ribonuclease A 1mg/mL2. Cytochrome c 1 mg/mL3. Transferrin 1 mg/mL4. Bovine Serum Albumin

1 mg/mL5. Concanavalin A

1 mg/mL6. Ovalbumin 1 mg/mL

Conditions: A) 0.1%Trifluoroacetic Acid in Water; B) 0.1% Trifluoroacetic Acid in Acetonitrile. Linear Gradient 20-60% B in 1.2 min. Ambient. 2 mL/min, 20 µL, UV at 215 nm.

The HxSil silica-based C18 and C8 columns show better resolution than the polymer-based PRP-1 and -Infinity columnsfor protein separations. The polymeric PRP-1 and -Infinitycolumns are used for protein purification when columncleaning/chemical sterilization is needed. Cleaning with acid orbase does not harm these polymeric columns. The PRP-Infinitycolumn (Figure 16) is well suited for the fast (less than 2 minute)separation of proteins.

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PeptidesFigures 17 through 19 illustrate the separation of a five componentdecapeptide standard on three Hamilton reversed phase columns.

The peptide standard contains the following five decapeptides;1. NH2-Arg-Gly-Ala-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide2. Acetyl-Arg-Gly-Gly-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide3. Acetyl-Arg-Gly-Ala-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide4. Acetyl-Arg-Gly-Val-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide5. Acetyl-Arg-Gly-Val-Val-Gly-Leu-Gly-Leu-Gly-Lys-amide

Figure 17Five Peptide Standards PRP-1, 4.1 x 150 mm, 5 µm (P/N 79444)


Conditions: A) 0.1%Trifluoroacetic Acid; B) 50% Acetonitrile, 0.09%Trifluoracetic Acid, Linear Gradient 0 -100% B in 50 min. Ambient. 1 mL /min, 100 µL, UV at 215 nm.


Conditions: A) 0.1% Trifluoroacetic Acid; B) 50% Acetonitrile, 0.09% Trifluoracetic Acid, Linear Gradient 0 -100% Bin 50 min. Ambient. 1 mL /min, 100 µL, UV at 215 nm.

Conditions: A) 0.1%Trifluoroacetic Acid; B) 50% Acetonitrile, 0.09% Trifluoracetic Acid, Linear Gradient 0 -100% B in 50 min. Ambient.1 mL /min, 100 µL, UV at 215 nm.

It is useful to note the unique selectivity of the PRP-1 polymeric,poly(styrene-divinylbenzene) HPLC column. It completely separatesall five peptide standards. PRP-1 polymeric columns have the addedadvantage of pH stability from 0 to 14, and they are pressure stable to 5,000 psi. The HxSil C18 and C8 columns separate three of thestandards and only partially resolve the first two peptide standards.None of the C18 silica columns tried (Discovery®, Vydac®, and Nova-Pak®) separated all five peptide standards.

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Technical DetailsHamilton HxSil C18 columns are functionalized mono-merically with octadecyldimethylchlorosilane and endcapped with trimethylchlorosilane. C8 columns arefunctionalized with octyldimethylchlorosilane and endcapped with trimethylchlorosilane.

EfficiencyThe tangent method is used to determine column efficiency. This method is common to a variety of column manufacturers and is easily performed by theuser. Typical efficiencies are on the order of >10,000plates per column for a 150 mm long column.

Reproducibility and SymmetryHxSil C18 and C8 columns are manufactured to provide reproducible retention of your compound. Tight manu-facturing controls and extensive characterization of thebase silica enable us to manufacture C18 columns withreproducible retention. Just look at the results of fourbatches of HxSil C18.

% Carbon 16.6 17.6 17.9 17.9µmol ligand/m2 3.0 3.1 3.2 3.2Particle Size 5 µm 5 µm 5 µm 3 µm

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USP MethodsThe Hamilton HxSil C18 column meets the requirements of a USP L1 column and the HxSil C8 column meets the requirements foran L7 column. Figure 5 illustrates the analysis of tetracycline.

Figure 5Tetracycline by USP Method HxSil C18, 4.6 x 250 mm, 5 µm (P/N 79869)

Figure 6 Folic Acid and Methotrexateby USP Method HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)

Figure 7Oxytetracycline andTetracycline HxSil C8, 4.6 x 150 mm, 5 µm (P/N 79102)


Conditions: 680 mL 0.1NAmmonium Oxalate,270 mL Dimethylformamide, 50 mL 0.2 M DibasicAmmonium Phosphate pH 7.6. 1 mL/min, 10 µL, Isocratic. Ambient. UV at 280 nm.


Conditions: 9:1 0.126 MSodium Phosphate, dibasic,0.037 M Citric Acid pH 6.0 :Acetonitrile. 1.2 mL/min, 10 µL, Isocratic. Ambient. UV at 302 nm.


Conditions: 680 mL 0.1N AmmoniumOxalate, 270 mL Dimethylformamide, 50 mL 0.2 M Dibasic Ammonium Phosphate pH 7.6. 1 mL/min, 20 µL, Isocratic. Ambient. UV at 280 nm.

Figure 8Anticonvulsants HxSil C18, 4.6 x 150 mm,5 µm (P/N 79868)

1. Ethotoin 2 µg 2. Phenobarbital 2 µg3. Methsuximide 2 µg4. Diazepam 0.5 µg

Conditions: A) Water; B) Acetonitrile. Linear Gradient 20-60% B in 10 min. Hold at 60% B for 5 min. Ambient. 1 mL/min, 100 µL, UV at 254 nm.

Figure 6 demonstrates the separation of folic acid from methotrexateon a HxSil C18 “L1” column following the USP 24 recommended conditions.

AnticonvulsantsThree anticonvulsants and one antianxiety drug are separated in a 12 minute gradient run with only an acetonitrile, water mobilephase (see Figure 8). All four peaks have excellent symmetry,narrow band widths and good resolution.

AntibioticsThe separation of oxytetracycline and tetracycline using an HxSil C8 column is shown in Figure 7.

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RetentionHamilton HxSil C18 columns exhibit greater retention than most columns. This allows you to separate compounds that are not suffi-ciently retained on other C18 columns. Figures 1 and 2 illustrate theseparation of polycyclic aromatic hydrocarbons (PAH’s) on an HxSilC18 column versus a Nova-Pak® C18 column of the same length. Theflow rate has been reduced for the Nova-Pak C18 column to compen-sate for the smaller I.D. The Hamilton HxSil C18 column providesabout twice the retention for these compounds (23 minute run time

versus 11 minutes). The greater retention of the HxSil C18 column provides better separation of these PAH compounds.With the HxSil C18 column, peaks 2 through 6 (naphthalene, acenaphthylene, acenaphthene, phenanthrene, anthracene) arebaseline resolved. The Nova-Pak C18 column does not fully separate peaks 2 and 3; peaks 5 and 6 are only partiallyresolved. Peaks 9 and 10 are better separated on the HamiltonHxSil C18 column.

Figure 1Polycyclic Aromatic Hydrocarbons (PAH’s)HxSil C18, 4.6 x 150 mm, 5 µm (P/N 79868)


Conditions: 7:3 Acetonitrile : Water.2 mL/min, 50 µL, Isocratic. Ambient. UV at 254 nm.

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Figure 2Polycyclic Aromatic Hydrocarbons (PAH’s)Nova-Pak C18, 3.9 x 150 mm, 5 µm


Conditions: 7:3 Acetonitrile : Water.1.4 mL/min, 50 µL, Isocratic. Ambient. UV at 254 nm.

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Figure 3Eight Small Molecules HxSil C18, 4.6 x 150 mm,5 µm (P/N 79868)

Figure 4Eight Small Molecules Inertsil ODS-3, 4.6 x 150 mm, 5 µm





SelectivityEach manufacturer’s reversed phase column is different. Choosing the best column to separate your sample can be difficult. Hamiltonmanufactures both silica-based and polymeric reversed phase HPLCcolumns, providing you with a wide range of column retention selectivities and performance benefits. The applications which follow,demonstrate the performance characteristics of Hamilton silica-basedreversed phase columns.

Small MoleculesFigures 3 and 4 compare Hamilton HxSil C18 and Inertsil® ODS-3 columns of the same length and particle size. The HxSil C18 columnprovides slightly longer retention (1 minute), and better separation ofpeaks 7 and 8 (amylbenzene and o- terphenyl). On the Inertsil ODS-3column these two compounds coelute.

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The HPLC Performance Report for each lot of HxSil C18 and C8 support includes information for the parameters listed below.

Silica Lot NumberParticle Size dv 50Particle Distribution dv 90/10Pore VolumeSilica Specific Surface AreaMetals

SodiumIronAluminum

Hamilton Company’s new HxSil silica-based reversed phase C18 and C8 HPLC columns provide greater retention of compoundsthan other C18 and C8 columns. This allows you to separatecompounds that are not adequately retained on other reversedphase columns. When you combine HxSil columns with our selection of polymer-based reversed phase columns you have awide range of column retention selectivity, and performance characteristics from which to choose.

At Hamilton we believe that your separation requirements dictate the type of column needed. Whether it’s a silica-based or polymer-based reversed phase column we are ready to help. Call us at 800-648-5950 for expert technical assistance and FREE method development. We understand that sometimes the only way to know if an HPLC column will separate your compound is to see your sample run on that column.

Each batch of HxSil C18 and C8 support is tested to determineretention and tailing of six model compounds. This mixturedeveloped by Steffeck et al.1 is used to determine the suitability of the C18 bonded phase and behavior of the underlying silicatoward neutral, hydrophobic, acidic and basic compounds. Thetailing factor for each compound is measured at 5% of peakheight (more demanding than measurment at 10% of peakheight).

The purpose of each probe is as follows: Uracil is a void volume marker.Pyridine is a basic compound used to test the activity of silanols toward bases.Phenol is an acidic compound used along with pyridine to determine the underlying silica activity.N,N-Dimethylaniline is a second basic compound used to determine the activity of residual silanols toward bases.4-Butylbenzoic Acid is a second acidic compound used to test the activity of residual silanols toward acids.Toluene tests the hydrophobicity of the column.

Both the Hamilton C18 (Figure 20) and C8 (Figure 21) columns separate all six test probes without a tailing peak (a tailing peak isdefined as a peak with a tailing factor greater than 2.0). This demon-strates the suitability of Hamilton HxSil C18 and C8 columns for theanalysis of acids, bases and neutral compounds. Of the 86 columnstested in the Steffeck et al. paper, 53 (62%) of the columns had 1 ormore of the test probes listed as a tailing peak.

Test Probes

Bonded Phase Lot NumberLigandCarbon Loading %µmol Ligand/m2

Capping Agent







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Document No. L 80065 Rev B © Hamilton Company 8/09

TrademarksThe following company trademarks have been used in this brochureDiscover y® - Sigma-Aldrich Nova-Pak ®- WatersInertsil ® - GL Sciences PRP ®, HxSil TM - Hamilton Company

References1. Robert J. Ste�eck, Susan L. Woo, Raymond J. Weigland and James M. Anderson, “A Comparison of Silica-Based C18 and C8 HPLC Columns to Aid Column Selection” LG* GC 13, 9 (1995): 720-726.

http://www.hamiltoncompany.com/hplcSales/Support USA 1-888-525-2123

Hamilton CompanyP.O. Box 10030Reno, Nevada 89520-0012Toll-Free 800-648-5950Tel: +1-775-858-3000Fax: +1-775-856-7259e-mail: [email protected]://www.hamiltoncompany.com

9 0 01

DNA PURIFICATION BY REVERSED PHASEAND ANION EXCHANGE HPLC

Hamilton Company offersthree HPLC columns forthe separation and purifi-cation of synthesizedDNA. Two for reversedphase gradient elutionchromatography: PRP®-1,PRP-3 and one for anionexchange gradient elution chromatography:PRP-X600.

Reversed Phase HPLC Purification of DNAHamilton polymeric reversed phase columns are copolymers of styrene and divinylbenzene. PRP-1and PRP-3 columns provide a number of benefitsfor anyone purifying synthesized oligomers byreversed phase HPLC. These include:

Excellent sample recovery, of greater than 95%.

High sample capacity, 10 mg or more for a 4.1 x150 mm column.pH stability for purification of DNA with secondary structure at pH 12.7.Long life, the packing is stable to a wide range of solvents and mobile phases.

Wide ApplicationPRP-1 and PRP-3 reversed phase columns areyour best choice for purification of syntheticDNA, RNA,and chimeric DNA-RNA oligomers.With PRP-1 and PRP-3 polymeric reversed phasecolumns you can purify (5’-monomethoxytrityl1,5’-dimethoxytrityl1,2, fluorescein, phosphoroth-ioate4, tert-butyldimethylsilyl5,7, and N-Acetyl-2-aminofluorene3,10) protected and deprotectedoligomers with excellent recovery. Oligomerswith secondary structure, e.g.hairpin loops and guanine rich sequences, are easily purifiedas well.

Excellent Sample RecoveryRecovery of trityl-on DNA 1 and chimeric DNA-RNA oligomers is greater than 95%7 and recov-ery of oligomers containing secondary structure is greater than 90%2 with PRP-1 columns.

This is possible because the polymeric(poly(styrene-divinylbenzene)) packing in PRP-1and PRP-3 columns are free from acidic silanolgroups. C18 columns typically recover only 50-80% of protected DNA because the silanolgroups on the column irreversibly bind the protected DNA or hydrolyze the dimethoxytrityl(DMT) protecting group during purification.

High Sample CapacityThe sample capacity of a 4.1 x150 mm PRP-1column was determined to be 10 to 25 mg ofDMT protected DNA per run by Ikuta et al.1.When compared to 1 mg/run for a C18 columnof the same size, it is readily apparent that PRP-1and PRP-3 are the columns of choice for protect-ed oligodeoxynucleotide purification. Loadingsup to 500 mg have been obtained on a 21.5 x250 mm column. A column’s sample capacity isdirectly proportional to its volume. For example,a 4.1 x 150 mm analytical column has approxi-mately 2.0 mL of volume, while a 21.5 x 250 mmpreparative column has a volume of 90.8 mLs. Ifyou can purify 10 mg on the analytical columnyou will be able to purify 45 times that amountor 450 mg,on the preparative column.

2

Another key to reproducible chromatography andscale-up is column re-equilibration with 10 ormore column volumes of mobile phase “A” priorto sample injection and gradient elution. Sincemost analytical HPLC pumps have an upper flowrate limit of 10 mL/min, the retention time of theoligomer will increase when using a larger column without increasing the flow rate propor-tionally. The following table can be used todetermine the expected oligomer capacity of acolumn and gives you an idea of the equilibra-tion volumes recommended for gradient elution.It is best to determine a column’s sample capacity using an analytical column, beforedeciding which preparative column to purchase.Other analytical, preparative and semipreparative column sizes are available, contact the HamiltonCompany.

Column Column EquilibrationSize Volume Volume

4.1 x 50 mm 0.7 mL 7 mL4.1 x 150 mm 2.0 mL 20 mL4.1 x 250 mm 3.3 mL 33 mL7.0 x 305 mm 11.7 mL 117 mL10.0 x 150 mm 11.8 mL 118 mL10.0 x 250 mm 19.6 mL 196 mL21.5 x 150 mm 54.4 mL 544 mL21.5 x 250 mm 91 mL 910 mL50.8 x 250 mm 507 mL 5,070 mL101.6 x 250 mm 2027 mL 20,270 mL

Semipreparative and preparative purifications(200 optical density units or greater) are bestmeasured at 297nm. This prevents saturation ofthe detector. Analytical separations are bestmonitored at 260 nm.

Long Column LifeThe poly(styrene-divinylbenzene) packing in PRP-1 and PRP-3 columns is stable. This stabilityprevents column degradation by any of thepurification methods mentioned in this brochure,assuring you reproducible separations and longcolumn life. The inertness of the stationary phasealso contributes to the exceptional recovery(>95%) of protected and deprotected oligomers.

Example Purification ProtocolA common DNA purification protocol for DMT-onoligomers follows:1. Load the DMT-on oligomer into the HPLC

injector after it has been cleaved from the CPG support and prior to detritylation.

2. Using mobile phases consisting of:A) 100 mM Triethylammonium Acetate pH 7.5;B) Acetonitrile. Run a gradient of 20-30%B

in 10 minutes (see Application #4). The order of elution is as follows:failure sequences,

oligomer without the dimethoxytrityl (DMT) then the purified DMT-on oligomer elutes.

3. The DMT-oligomer is collected and treated with 500 µL of 80% acetic acid for 30 minutes. The acid treatment removes the acid labile DMT protecting group from the oligomer. Since DNA is not stable in acidic solutions, we recommend you neutralize the solution with careful addition of Ammonium Hydroxide solution.

4. If needed, the detritylated oligomer can be repurified on the same column using the following isocratic conditions:9:1 100 mM Triethylammonium Acetate, pH 7.5/Aceton-itrile. The detritylated oligomer elutes quickly (see Application # 5).

5. The detritylated oligomer is collected and desalted if desired.

For additonal information, please consult the references listed at the end of the Mobile Phasessection.

Application #1PRP-1,4.1 x 250 mm





Isocratic Scaleup, 101.6 mm ID Conditions:0.05M Citric Acid,pH 4.20,160 mL/min,Ambient,UV at 254 nm.

Isocratic Scaleup, 21.5 mm IDConditions:0.05M Citric Acid,pH 4.20,7.16 mL/min,Ambient,UV at 254 nm.

Isocratic Scaleup, 4.1 mm IDConditions:0.05M Citric Acid,pH 4.20,0.26 mL/min,Ambient,UV at 254 nm.

0 10 20 30

0 10 20 30

0 10 20 30

1

1

1

2

3

2

3

2

3


Easy Purification ScaleupChanging from an analytical column to a semi-prep or preparative column is easy. Applications#1,2 and 3 illustrate the reproducible scaleupfrom a 4.1 mm column to a 101.6 mm ID column.


1. Failure Sequences

2. 5’-DMT 16mer


Repurification of a Deprotected 16mer Conditions: 9:1 100 mM Triethylammonium Acetate pH 7.5 : A c e t o n i t r i l e. 2.0 mL/min, A m b i e n t .UV at 260 nm.

DMT-on 16mer PurificationConditions: A) 100 mM Triethylammonium Acetate pH 7.5;B)Acetonitrile. Linear Gradient 20-30% B (0-8 min),30-80% B(8-15 min).2.0 mL/min,

1. Deprotected 16mer

0 5 10

0 1 2 3 4 5

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1

2

3

Mobile Phases A wide variety of mobile phases and sample preparation conditions can be used withPRP-1 and PRP-3 reversed phase columns forpurification of protected and deprotected DNAoligomers. The information that follows represents a summary of the LiteratureReferences listed at the end of this brochure.

Purification of DMT-on and DMT-off DNAMOBILE PHASE #1Buffer A) 100 mM Triethyammonium

Acetate pH 7.5 Buffer B) AcetonitrileAdvantages: DNA is typically purified with the

DMT group attached,then deprotected and repurified with a shallower gradient.

Disadvantages: None mentionedLiterature Referance:#9

Purification of DMT-on DNA withSecondary StructureMOBILE PHASE #1Buffer A) 10 mM Ethylenediammonium

Acetate pH 7.6 Buffer B) 1:110 mM Ethylenediammonium

Acetate pH 7.6 :AcetonitrileAdvantages: High column capacity of 10 mg for

the 4.1 x 150 mm size.Purify DNA with self complementary sequences.

Disadvantages: Column temperature is 60°C.Literature Reference #1

MOBILE PHASE #2Buffer A) 50 mM Sodium Hydroxide pH 12.7 Buffer B) 1:1 50 mM Sodium Hydroxide :

Acetonitrile

Sample Prep: None except some very stable structures must first be treated with formaldehyde.

Advantages: Straight forward,easy to use method.

Disadvantages: Column must be heated to 60°C. Method will not work with modified,base labile DNA.

Literature Reference #2

MOBILE PHASE #3Buffer A) 10 mM Potassium Phosphate pH 7Buffer B) 7:1:2 V:V:V Acetonitrile : Methanol :

10 mM Potassium Phosphate pH 7.Sample Prep: Up to 10 OD of crude DNA

were predissolved in 400 uL of 10 mM Potassium Phosphate pH 7. Then 100 uL of formamide is added, vortexed and heated in a water bath at 90°C for 3 minutes and injected.

Advantages: These buffers and the sample preparation allow the purification of strongly interacting DNA with guanine-rich or self-complementary regions without column heating or high pH.This method is also useful for purification of chemically modified or adduct containing DNA. These buffers are useful for purification of samples containing less than 5 to 10 OD’s.

Disadvantages: More laborious sample prepLiterature Reference: #8

MOBILE PHASE #4Buffer A) 50 mM Triethylammonium Acetate

pH 7 :5% Acetonitrile Buffer B) 50 mM Triethylammonium Acetate

pH 7 : 50% AcetonitrileSample Prep: Desalted DNA was predissolved in

20 µL of Deionized Water. Then 80 µL of

formamide was added, vortexed and heated in a water bath at 90°C for 3 minutes.Then chilled in an ice bath. 400 uL of chilled 50 mM Triethylammonium acetate pH 7 was added,vortexed and injected.

Advantages: These buffers and the sample preparation allow the purification of strongly interacting DNA with guanine rich or self-complementary regions without column heating or high pH.This method is also useful for purification of chemically modified or adduct-containing DNA. These buffers are also useful for largerscale purifications (higher than 5 to 10

OD’s).Disadvantages: More laborious sample prep

Literature Reference: #8

Purification of DMT-on DNA for NMRMOBILE PHASE #1Buffer A) 10 mM Potassium Phosphate pH 7.0 Buffer B) 7:1:2 V:V:V Acetonitrile : Methanol :

10 mM Potassium Phosphate pH 7.0Advantages: This method avoids the use of

amines which can cause difficulty with NMR analysis. It also eliminates the need for the usual ion exchange purification step.

Disadvantages: None mentionedLiterature Reference #3

1. FailureSequences

2. 5’-DMT 36mer


DMT-on 36mer with Secondary StructureConditions: A) 50 mM Sodium Hydroxide pH 12.7;B) 1:1 50 mM Sodium Hydroxide :Acetonitrile.Linear Gradient 0% B (0-2 min),0-50% B (2-25 min).2.0 mL/min,60°C. UV at 260 nm.



2. 5’-DMT 30mer

DMT-on 30mer PurificationConditions: A) 100 mM Triethylammonium Acetate pH 7.5;B) Acetonitrile. Linear Gradient 20-30% B (0-8 min),30-80% B(8-15 min).2.0 mL/min,Ambient.UV at 260 nm.

0 2 4 6 8 10 0 10 20

1

21

2

Short Tandem Repeat PrimersConditions: A) 100mM TEAA,0.1 mM EDTA pH 7; B) 75:25100mM TEAA,0.1 mM EDTA pH 7 :Acetonitrile.Linear 0.2 Gradient 15-50% B in 40 min. Temperature Gradient80-90ºC in 16.7 min.Hold at 90ºC. 1 mL/min.UV 260 nm.

Application # 6PRP-3,4.6 x 50 mm

0 10 20 30 40

4

Purification of DMT-on DNA with Secondary Structure and On-Column DMT DeprotectionMOBILE PHASE #1Buffer A) 15% Acetonitrile in 100 mM

Tetraethylammonium HydroxideBuffer B) 15% Acetonitrile in 100 mM

Triethylammonium Bicarbonate pH 7.0Buffer C) 0.5% Trifluoroacetic Acid Buffer D) AcetonitrileAdvantages: Purify large amounts of DNA for

NMR,x-ray crystallography and antisense studies. The alkaline denaturing conditions can purify 10 µmole scale synthesis.Detritylation on the HPLC column and load the sample in ammonium hydroxide. Good purity as determined by CE and NMR.

Disadvantages: None mentionedLiterature Reference: #6

Purification of Phosphorothioatemodified DNAMOBILE PHASE #1Buffer A) 100 mM Potassium Phosphate with

2 mM Tetrabutylammonium Phosphate,pH 7.0

Buffer B) Acetonitrile Sample Prep: Plasma and urine samples were

prepared with phenol extraction procedure.Advantages: Sharper peaks than silica-based

column under identical conditionsDisadvantages: None mentioned Literature Reference: #4

Purification of 2-(acetylamino)fluorene Modified DNAMOBILE PHASE #1Buffer A) 10 mM Potassium Phosphate

pH 7.0 Buffer B) 7:1:2 V:V:V Acetonitrile :

Methanol : 10 mM Potassium Phosphate pH 7.0

Advantages: This method avoids the use of amines which can cause difficulty with NMR analysis. It also eliminates the need for the usual ion exchange purification step.

Disadvantages: None mentionedLiterature Reference #10

Purification of Fluorescein Modified DNAMOBILE PHASE #1A) 20 mM Sodium Acetate pH 6.7 B) Acetonitrile(see application #9)

Purification of RNA and DNA/RNAChimersMOBILE PHASE #1Buffer A) 10 mM Tetrabutylammonium

acetate, 1 mM Tetrabutylammonium phosphate pH 7.5

Buffer B) 8:2 (V:V) 10 mM Tetrabutylammonium acetate, 1 mM Tetrabutylammonium phosphate pH 7.5, Acetonitrile :Water Column Temperature:60°C

Advantages: Long column lifetime. Very good recoveries of the sample (95%).PRP-1 columns are stable to the tetra-n-butylammonium fluoride used in the sample preparation. Silica-based columns are not. The level of purifica-tion possible with PRP-1 is comparableto that which might be achieved by preparative gel electrophoresis but PRP-1 is able to purify much larger samples more rapidly. Now prepare relatively large amounts of high purity chimeric ribozymes without PAGE purification.

Disadvantages: Column must be heated to 60ºC

Literature Reference: #7

Purification of RNAMOBILE PHASE #1Buffer A) 100 mM Trimethylammonium

acetate pH 7.0Buffer B) Acetonitrile

Column Temperature:60°CAdvantages: PRP-1 columns are more stable to

continous heating than silica-based C18 columns. Superior recovery of RNA versus other columns. A volatile buffer eliminates the need for desalting. The Trimethyl-ammonium salt is considerably easier to remove by lyophilization than the Triethylammonium salt.

Disadvantages: Column must be heated to 60ºCLiterature Reference: #5

Literature References1.Ikuta, S.;Chattopadhyaya,R.;

Dickerson,R. Analytical Chemistry,1984, 56, 2253.

2.Germann,M.W.; Pon,R.T,; van de Sande, J.H. Analytical Biochemistry,1987, 156, 399.

3.Huang, G.;Krugh, T.R. AnalyticalBiochemistry. 1990, 190, 21.

4.Biegelow, J.C.;Chrin,L.R.;Mathews, L.A.;McCormack, J.J.J Chrom., 1990, 533, 133.

5.Khare, D.;Orban, J. Nucleic Acids Research, 1992, 20, 5131.

6.Hobbs, F.W.;Yarem, J.A.BioTechniques 1993, 14, 584.

7.Swiderski, P.M.;Bertrand,E.L.;Kaplan, B.E. Analytical Biochemistry,1994, 216, 83.

8.Arghavani,M.B.;Romano, L.J.Analytical Biochemistry, 1995, 231, 201.

9.Reddy, D.M.;Iden, C.R. American Laboratory, 1995, 15, 15.

10.Zhou, Y.;Romano, L. Biochemistry,71993, 32, 14043.


2. Fluorescein 12mer


Fluorescein Modified 12merConditions: A) 20 mM Sodium Acetate pH 6.7;B) Acetonitrile.Linear Gradient 0-50% B (0-20 min). 1.0 mL/min,Ambient.UVat 260 nm.

0 6 12

1

2

5

Reversed Phase Column Ordering InformationPRP-1, 100Å Reversed Phase HPLC Columns

I.D. x Length 5 µm 7 µm 10 µm

2.1 x 50 mm 79554

2.1 x 100 mm 79790

2.1 x 150 mm 79366 79480

2.1 x 250 mm 79390 79391

4.1 x 50 mm 79443

4.1 x 100 mm 79479 79565

4.1 x 150 mm 79444 79529 79425

4.1 x 250 mm 79820 79422 79427

4.6 x 100 mm* 79558

4.6 x 150 mm* 79423 79351

I.D. x Length 5 µm 7 µm 10 µm 12-20 µm

4.6 x 250 mm* 79571 79380 79381

7.0 x 100 mm 79495 79713

7.0 x 305 mm 79795 79426

10.0 x 50 mm 79367

10.0 x 100 mm 79355 79499

10.0 x 250 mm 79531 79496

21.5 x 250 mm 79352 79478 79428

50.8 x 250 mm 79567 79493

101.6 x 250 mm 79525

* PEEK hardware

PRP-3, 300Å Reversed Phase HPLC Columns


2.1 x 150 mm

4.1 x 150 mm

4.1 x 250 mm

4.6 x 150 mm*

4.6 x 250 mm*

7.0 x 305 mm

10.0 x 250 mm

21.5 x 250 mm

* PEEK Hardware

PRP-3 Semiprep/PreparativeGuard Column For Steel Columns***


Replacement 79124Cartridges (2/pk)

*** Semiprep/preparative guard columns for steel columns are 4.6 x 20 mm

PRP-3 Analytical Guard Columns For Steel Columns* For PEEK Columns**

Starter Kits (1 holder, 2 cartridges)

ReplacementCartridges (5/pk)

* Analytical guard columns for steel columns are 2.3 x 20 mm

PRP-1 Semiprep/Preparative Guard ColumnFor Steel Columns***


Replacement 79122Cartridges (2/pk)

*** Semiprep/preparative guard columns for steel columns are 4.6 x 20 mm

PRP-1 Analytical Guard ColumnsFor Steel Columns* For PEEK Columns**

Starter Kits 79447 79317(1 holder, 2 cartridges)

Replacement 79445 79318Cartridges (5/pk)

* Analytical guard columns for steel columns are 2.3 x 20 mm ** Analytical guard columns for PEEK columns are 3.0 x 8.0 mm

.

79461 79393

79454 79395 79392

79794

79574

79382

79466

79469

79468

79526

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Anion Exchange Purification of DNAHamilton PRP-X600 anion exchange HPLCcolumns separate DNA oligomers according to charge. Controlled porosity provides excellentseparation of n-1 oligomers from the full length product. See application #11 for the separationof a DMT-on antisense, phosphorothioate 18mer from the failure sequences. PRP-X600 columnswork best when heated to 85°C.

Sample Capacity The 50 mm long 4.6 mm I.D. columns have asample capacity of approximately 1 mg.150 x4.6 mm long columns have a sample capacity of about 3 mg.

Compatible Mobile PhasesPRP-X600 weak base anion exchange columnsare polymeric,stable from pH 1 to 11 andcompatible with organic solvents.

Changing Mobile Phases andEquilibration After GradientSeparationEquilibration of the PRP-X600 column requiresapproximately 10 column volumes. Followingeach gradient analysis, reequilibrate the columnwith Buffer A to ensure reproducible sample retention.

Application #10PRP-X600,4.6 x 50 mm

Application #11PRP-X600,4.6 x 50 mm

pBR322 DNA Fragment Conditions: A) 20 mM TRIS, 1 mM EDTA pH 9.0;B) 1N Sodium Chloride in 20 mM TRIS, 1 mM EDTA.Linear Gradient 60-67.5% B (0-15 min),67.5-75% B (15-45 min).1.0 mL/min,Ambient.UV at 260 nm.

0 10 20 30 40

DMT-on Phosphorothioate Antisense 18mer Conditions: A) 85:15 100 mM TRIS pH 8.0 :Acetonitrile;B) 85:15 100 mM TRIS pH 4.0,2.5 M Lithium Chloride :Acetonitrile. Linear Gradient 0-100% B in 10 min.2.0 mL/min. 85°C. UV at 260 nm

Anion Exchange HPLC Purification of DNA

0 10 20 30

1. DMT-on phosphorothioate antisense 18mer

1

PRP-X600 Anion Exchange HPLC ColumnsI.D. x Length 7 µm

4.6 x 50 mm* 79360

4.6 x 150 mm* 79189

7.5 x 250 mm 79492

* PEEK Hardware

PRP-X600 Guard Columns For PEEK Columns

Starter Kits (1 holder, 2 cartridges) 79361


7

Hamilton HPLC supports for the separation and purification of DNA are available as bulk resin or prepacked into analytical,semi-preparative and preparative columns. Chromatographic capacity and efficiency tests are conducted on both bulk and packed column resins to ensure product integrity and reproducibility. Samples from different production lots can be purchased for method/process validation. For ease of ordering,specify the quantity of resin needed in grams.Quantities from one gram to tens of kilograms are available. One gram of resin is roughly equivalent to 2.5 cc’s of volume.

Visit the Hamilton Company web site at http://www.hamiltoncompany.com for technical specifications.

Bulk Resin

SupportType of Support Material Capacity Pore Size 5 µm 7 µm 10 µm 12-20 µm 30-50 µm 50-75 µm

PRP-1 PSDVB* N/A 100Å 79578 79579 79580 79581 79582 79583Reversed Phase

PRP-3 PSDVB* N/A 300Å 79701 79702Reversed Phase

PRP-X600 Polydimethyla- 1.6 Superficially 79192 79597 79598 79599Anion Exchange midopropylme- meq/gm porous

thacrylamide

* PSDVB is poly(styrene-divinylbenzene).Bulk resin is sold by the gram.

Bulk Resin Ordering Information

Use Hamilton SPE cartridges, barrels, and 96-wellplates for rapid and easy purification of synthe-sized DNA.

Excellent Purity - PRP-1 cartridges providepurity comparable to that obtained with reversedphase HPLC. Depending on your needs, you havethe choice of a purified protected oligomer or apurified deprotected oligomer when you usePRP-1 cartridges.

See Figure 1 for an electropherogram of a 27merpurified with a PRP-1 cartridge.

Higher Sample Capacity - One PRP-1 cartridge will purify 20 OD A260 units from a 0.2 µmole synthesis. Apply your sample once,at the rate of 1-2 drops per second for optimalbinding. PRP-1 cartridges do not require multipleapplications.Low Backpressure - Cartridges have very low

SPE Cartridges, Barrels and 96-well plates

backpressure, so loading your sample is easier.Tight controls on particle size ensure consistentbatch to batch performance.

Cartridges are designed for a single use, butmay be used again for purification of the sameoligonucleotide. Cartridges can be used with asyringe or vacuum manifold to load, wash, andelute your samples. Barrels are designed for usewith a vacuum manifold.

SPE Purified OligomerElectropherogram of a 27mer purified with a Syringe Cartridge

Hamilton Company4970 Energy WayReno, Nevada 89502 USAToll-Free: 800-648-5950Telephone: +1-775-858-3000Fax: +1-775-856-7259e-mail: [email protected]

Hamilton Bonaduz AG Via Crusch 8CH -7402Bonaduz/SwitzerlandTelephone: +41-(0)81-660-60-60Fax: +41-(0)81-660-60-70e-mail: [email protected]

R

Document No. L80056 (Rev. C) © Hamilton Company 04/09

Web site: http://www.hamiltoncompany.com

0 25 30 35Minutes

SPE Ordering InformationPRP-1 Syringe CartridgesPart # Description Bed Volume Application

79530 Syringe Cartridge 50/pk 150 mg 0.2 µmole

79537 Syringe Cartridge 10/pk 330 mg 0.3-0.5 µmole

79559 Syringe Cartridge 10/pk 600 mg 1.0 µmole

PRP-1 Barrels (for use on vacuum manifolds)Part # Description Bed Volume Application

79342 Barrel,1 mL,50/pk 100 mg 0.2 µmole

79566 Barrel,1 mL,50/pk 150 mg 0.2 µmole

79345 Barrel,6 mL,10/pk 1 gm 1.5 µmole



96 Well PlatesPart # Description Bed Volume Application

79164 96 Well Plate 150 mg/well 0.2 µmole

Electropherogram courtesy of Lolita Bland,University of VirginiaPRP is a registered trademark of Hamilton Company

liquid chromatography - cosela 1317638748.pdf · 81 4-amino-n10- methylpteroglutamic ... 100 [sar1,...

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