practical examples of method translation 040209 … › cs › library ›...

Practical Applications of Method Translation Using g

the Agilent Method Translation Tool

Rita SteedInside Application EngineerAgilent TechnologiesLife Sciences and Chemical AnalysisApril 2, 2009

Practical Applications of Method Translation Using the Agilent Method Translation Tool

Title4/1/2009

Objectives

• Demonstrate Agilent Method Translation tool – For fast, easy, and successful method transfer to , y,

smaller volume columns• Review important method variables

I ti– Isocratic– Gradient

• Review method transfer examples using the• Review method transfer examples using the Agilent Method Translation Tool


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Successful Method Translation

• Separation Goals and Method Performance Criteria• Isocratic separations

InstrumentationInstrumentationWhich instrument and method parameters afford optimal resultsConsiderations for successful implementationA il t M th d T l t f i ti tiAgilent Method Translator for isocratic separations

• Gradient separationsGradient retention parametersInstrument considerationsAgilent Method Translator for gradient separations


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Separation Goals and Method Performance Criteria

Separation Goals and System Suitability• Resolution: ≥2

Method Performance CriteriaAccuracy

• Peak shape: USP Tf close to 1 (<2)

• Injection Repeatability: areas, Tf, etc. (RSD 0.1 - 0.25%)

Precision• Ruggedness

– Repeatability

• Absolute retention: 1< k>10

• Relative Retention: α or k2/k1

• Signal to Noise Ratio: >10

– Intermediate precision– Reproducibility

• RobustnessSelectivity/Specificity

• Signal-to-Noise Ratio: >10

AVOID THESE for System Suit. Criteria

Linearity

Range

Quantitation Limit (LOQ 10x S/N)Column efficiency (theoretical plates)

Absolute retention

Quantitation Limit (LOQ, 10x S/N)

Detection Limit (LOD, 3x S/N)


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An Approach for Isocratic Method Translation

• Assess current method performance and parameters

• Set performance goals for method to be translated

• Determine column geometry for necessary efficiency

• Instrument needs vs. method performance goals Depend on requirements for column size and particle size to get desired R– Depend on requirements for column size and particle size to get desired Rs

• Instrument ECV, detector data rate• System pressure limitations

Adj t i j ti l f ll l l• Adjust injection volume for smaller column volume

• Assess injection repeatability and sample solvent composition robustness

• Adjust flow rate vs. system max. pressure relative to method performance goal for analysis time.


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Isocratic Method: Document current method performance, parameters, and instrument configurationCurrent Method Performance• Limiting resolution for critical pair(s)• Peak shape(s) (USP T )

Method Parameters

• Column length, id and particle size• Peak shape(s) (USP Tf)• Injection repeatability• Signal-to-Noise ratio

• Flow rate

• Mobile phase composition (viscosity)

• Column temperature

Instrument Configuration• Extra Column Volume

Column temperature

• Injection volume

• Sample concentration and sample l t iti• Tubing ID and length

• Flow cell volume• Detector data rate

solvent composition

• Nominal backpressure

• Flow cell pathlength• System maximum pressure


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Isocratic Method ExampleSituation:You have isocratic method for tocopherols developed for 4.6 mm i.d. columns in 150 mm length. Run time is ~14 min.

Pump: Agilent 1100 quaternary systemAutosampler: Standard autosamplerTCC: 1100 standardDetector: 1100 DAD, max. data rate 20 Hz

Typical setting, PW = 0.05 min.Flow Cell: 13 μL, 10 mm path lengthFlow Rate: 1.0 mL/min.Column temp. 23ºCGoals: Decrease run time and improve throughput (5X, if possible)

S l t d t (i li ll l id h tSave solvent usage and waste (implies smaller column id or shorter run at higher flow rate)

• Can anything be done to speed up these methods with existing equipment?

What modifications can be made and which are most important?


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• What modifications can be made and which are most important?

Isocratic method on Conventional ColumnTocopherols

Column: ZORBAX Eclipse XDB-C18Mobile Phase: 95% ACN: 5% WaterTemp: 23ºCRRHT

p

Conventional4.6 x 150 mm 5 μm

Flow Rate: 1 mL/minP = 37 bar

Temp: 23 CInjection volume: 1 uL

RRHT4.6 x 50 mm 1.8 μm

Flow Rate: 3 mL/minPressure = 229 bar

P = 37 barmAU

60

80

Rs ~ 4.4Rs ~ 5.2

20

40 1.7 min13.5 min

Sample: Vitamin E – α, β, γ-tocopherols in gel capE li XDB C18 i d fi t h i f th d

min0 2 4 6 8 10 12 140


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Eclipse XDB-C18 is a good first choice for many methods.

Assess Your Current Method

Assess your current method4.6 x 150 mm, 5 μm column

Questions to ask?What is the mobile phase composition?

1.0 mL/minRT last = 14 minutes

What is the current backpressure?Injection volume?Data rate/Peak width?What is your limiting resolution with current method?What size column can deliver the

l ti d?resolution you need?Can your current instrument be used with a shorter column with smaller particle size?Which changes in method parameters are necessary and can you get the same or similar performance and results?


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Efficiency Ranking of Various Column Geometries and Typical Backpressuresand Typical Backpressures

This RRHT column Replaces These Longer Columns50 mm, 1.8 μm 150 mm, 5 μm, 100 mm, 3.5 μm

100 mm, 1.8 μm 250 mm, 5 μm


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Tocopherol Method Translation

Current Method4.6 x 150 mm, 5 µm XDB-C18Viscosity of 95:5 ACN/water at 23ºC is ~0 43 cpViscosity of 95:5 ACN/water at 23 C is ~0.43 cpFlow Rate is 1 mL/minBackpressure is 37 barSt d d fl ll (13 L)Standard flow cell (13 µL)Standard 0.17 mm tubing throughoutLimiting Resolution ~4.4Peak Width required 0.1 minResponse Time = 2 sec or Data Rate = 2.5 Hz is adequate


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Agilent Method Translator


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Isocratic Method: Translation Tool 13 uL flow cell and 0.17 mm tubing

21.6 uL tubing vol + 13 uL

flow cell

Effective N hurt by EC vol.


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For isocratic runs the 2nd row must be set to same %B as row 1

Use 5 uL Flow Cell and 0.12 mm id Tubing

Improvement in N effective


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Adjust % max. pressure until desired flow

Speed Optimized at 3 mL/min

rate

Adjust to 3 mL/min

Click radioClick radio button to

allow % max pressure

adjustment


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Tocopherol Method Translation

Translated Method4.6 x 50 mm, 1.8 um RRHT XDB-C18C l l th i h t di i ith 1 8 ti l i 4 6 50 RRHTColumn length in shorter dimensions with 1.8 µm particles is 4.6 x 50 mm RRHTAt 1 mL/min expected backpressure is 79 bar + ~10 bar (a/s and flow cell) or ~90 barExpected run time will be ~1/3 of 14 minutes or 4.67 minutesTry 3 mL/min for run time of 1/9 of 14 min or 1 55 minTry 3 mL/min for run time of 1/9 of 14 min. or 1.55 min.Predicted pressure is 238 barLimiting resolution will be approximately the same (4.4) or 4.4 x SQRT(13043/12077) = 4.2, If no band broadening due to extra column volume or data rate.Standard DAD or MWD at fastest setting (20 Hz) with 0.17 mm id tubing adequate but not optimumChoose 0.12 mm i.d. tubing and 5 µL flow cell for better results


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Comparison of Conventional Isocratic Method vs. Translated Method at 3 mL/min

Column: ZORBAX Eclipse XDB-C18Mobile Phase: 95% ACN: 5% WaterTemp: 23ºCRRHT

Translated Method at 3 mL/min

Solvent used 15 mL

Conventional4.6 x 150 mm 5 μm

Flow Rate: 1 mL/minP = 37 bar

Temp: 23 CInjection volume: 1 uL

RRHT4.6 x 50 mm 1.8 μm

Flow Rate: 3 mL/minPressure = 229 bar

P = 37 barmAU

60

80

Rs ~ 4.4Rs ~ 5.2Solvent used 5.1 mL

20

40 1.7 min13.5 min

min0 2 4 6 8 10 12 140

Sample: Vitamin E – α, β, γ-tocopherols in gel capE li XDB C18 i d fi t h i f th d


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Eclipse XDB-C18 is a good first choice for many methods.

Flow Cells for RRLC

13 µl Standard Flow Cell: For highest sensitivityHigh-demanding quantitative work, e.g. analytical method development QA/QCanalytical method development, QA/QC

2 µl Micro Flow Cell: For highest resolutionUltra fast semi quantitative workUltra-fast semi-quantitative work, e.g. Screening Experiments, HT LC/MS/UV

5 µl Semi-micro Flow Cell: Best compromise of sensitivity and resolutionBest compromise of sensitivity and resolutionFor good quantitative and qualitative results, e.g. Screening, HT LC/MS/UV, Early Formulation Studies

Dimension Sensitivity* Resolution*

13 µl / 10 mm +++ +

* D d l ti l diti d l di i

5 µl / 6 mm ++ ++2 µl / 3 mm + +++


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* Depends on analytical conditions and column dimension

Effect of Detector Response Time on Fast Gradient Analyseson Fast Gradient Analyses

Response Time

Agilent 1100 DADA il t 1100 WPS ith ADVR

0.1 sec

0 21st peak = 1 2 sec Agilent 1100 WPS with ADVR

Column: Poroshell 300SB-C182.1 x 75 mm, 5 μm

Mobile Phase:A: 95% H2O, 5% ACN with 0.1% TFAB 5% H O 5% ACN ith 0 1% TFA

0.2 sec

0.5 sec

1 peak = 1.2 secAt 20 pts/sec = 24 pts/sec

B: 5% H2O, 5% ACN with 0.1% TFA

Flow Rate: 2 mL/min

Temperature:70°C

Detector: UV 215 nm

1.0 sec

1st peak = 1.2 secAt 5 pts/sec = 6 pts/sec

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9Time (min)

1.0

Piston stroke: 20

Sample:1. Neurotensin3. Lysozyme2. RNaseA 4. Myoglobin

2.0 sec

You may have to adjust the response rate of your detector for rapid peak detection.

High Speed LC with RRLC and RRHT ColumnsMaintaining Resolution at High Analysis Speed

PW=0 30se 80Hz versus 10Hz (20Hz) Data Rate

g g y p

80Hz

PW=0.30sec

80Hz versus 10Hz (20Hz) Data Rate• Peak Width: – 55% (– 30%)• Resolution: + 90% (+ 30%)• Peak Capacity: + 120% (+ 40%)• App Column Eff : + 260% (+ 70%)

Data Rate

Peak Width

Resolution Peak Capacity

80 Hz 0.300 2.25 6040Hz

PW=0.33sec• App. Column Eff.: + 260% (+ 70%)

40 Hz 0.329 2.05 55

20 Hz 0.416 1.71 45

10 Hz 0.666 1.17 29

5 Hz 1 236 0 67 16

20Hz

PW=0.42sec

PW=0 67sec 5 Hz 1.236 0.67 16

min0 1 0 2 0 3 0 4 0 50

10HzPW 0.67sec

5HzPW=1.24sec

Sample: Phenones Test MixColumn: Zorbax SB-C18, 4.6x30, 1.8umGradient:: 50-100%ACN in 0.3minFlow Rate: 5ml/min

min0.1 0.2 0.3 0.4 0.50 Flow Rate: 5ml/min

Translating Gradient MethodsTranslating Gradient Methods


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Advantages of Gradient Elution

Complex samples are analyzed in a single HPLC runAnalysis time can be reducedAnalysis time can be reducedPeaks elute with the same bandwidthMore peaks can be baseline resolved per unit timeMore peaks can be baseline resolved per unit time

– higher peak capacity than isocratic methodSignal-to-Noise ratios and LOD/LOQ are relatively the g ysame during a gradient run (barring ghost peaks, anomalies, etc.!)

peaks don’t broaden with increasing retention time as– peaks don t broaden with increasing retention time as they do in an isocratic separation)


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Resolution Relationship for Gradient Elution

VNVR ≈ N4

k*α

k* - represents the fact that k changes constantly during a gradient

87 tg F

S ( B) Vk* =

Δ%B = difference between initial and final % B valuesS = constant (≈ 4 for 100 - 500 Da)F = flow rate (mL/min.)tg = gradient time (min.)

S (Δ%B) Vmg

Vm = column void volume (mL)

Title of PresentationDate

Agilent Restricted

This Relationship Says that to Keep R l ti P k P iti i thRelative Peak Position in the Chromatogram Unchanged

• Column length • Decrease in tG or F

Any Decrease in Can be Offset by a Proportional

• Column length

C l l (i d )

Decrease in tG or F• Increase in ΔΦ

• Decrease in t or F• Column volume (i.d.)

( )

• Decrease in tG or F• Increase in ΔΦ

D i F• ΔΦ (same column) • Decrease in tG or F

k* =tG • F

k = S • ΔΦ • Vm

100% B Gradient Steepness AffectsRetention (k*) and Resolution

100% B

tg= 5

87tg Fk* =0% B

This equation

tg= 10 1/k* ∝ gradient steepness = b

ΔΦ Vm Sk

governs gradient retention and

selectivity

100% B

t =

ΔΦ = change in volume percent of B solvent (%)

S = property of sample compoundF fl t ( L/ i )

0% B

100% B

tg= 20

F = flow rate (mL/min.)tg = gradient time (min.)

Vm = column void volume (mL)0% B

0 10 20 30 40

tg= 400% B

• S ≈ 4–5 for small molecules• 10 < S < 1000 for peptides

and proteins


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0 10 20 30 40

Time (min)000995P1.PPT

Transferring a Gradient Method to a Small(er) Column

Examine the current method– Column length and i.d., particle size, N– Injection volume– Injection precision– Gradient program

• Initial Hold Time

It’s much easier to transfer a linear gradient than one with multiple

segments and hold times

• Linear gradient segments• Isocratic holds during gradient

– Delay Volume– Resolution of critical pair(s)– Backpressure

Can you trade excess resolution for time or can you get the same ffi i (N) ith h t l ?efficiency (N) with a shorter column? – Calculate critical pair resolution on shorter column(s) with smaller particle size(s)– Calculate expected pressure at one or more flow rates on shorter column


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Gradient Separations: Considerations When Translating Existing Gradient Methods

Isocratic Separations

Sample load; Vinj, analyte

Sample solvent strength

Gradient SeparationsSame as Isocratic Separations plus…

Sample solvent strength

Extra column volume

– Flow cell volume

Delay Volume– Same instrument (different pressures)– Different instrument (for example,

Capillary 1100 vs Binary 1100)– Injection volume– Tubing volume

Injector precision

Capillary 1100 vs. Binary 1100)Gradient Time

– Adjust relative to equation for gradient retentionInjector precision

– Can vary with Vinj

– Data RateToo fast too much noise

– Keep k* constant

Gradient Delay Time– Gradient delay time must be same as for

– Too fast, too much noise– Too slow, loss of N

larger column separation– Ratio of gradient volume/column volume

must be same as for larger column

Column Equilibration Time (Post Time)


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q ( )

Gradient Separations – What is Delay Volume?

Also known as

Dwell Volume

DelayVolume

• Delay Volume = volume from formation of gradient to the column• Behaves as isocratic hold at the beginning of gradient


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• Behaves as isocratic hold at the beginning of gradient.

Comparison of System Delay Volumes

1090 1050 1100 Quat 1100 Bin RRLC

Pump w/o mixerw/ mixer

300-5001050-1250

180-480600-900

800-1100n/a

800-1100n/a

100-120600-800

Std MixerUpchurch

Autosampler Standard

750

V (loop)

42081

300 + V (inj)

n/a

300 + V (inj)

n/a

327 + V

n/a

-Bypass

Column comp. StandardBypass

N/A

4.1 or 8.20

6.2

3 or 60

6.2

3 or 60

(inj)8

15 ul

5

3 or 60

0Min rangeMax range

304-5041058-1258

189-489906-1206

1203-14061242-1442 100-150*600-800

Speed

*competitive low volume LC – 150uL

Delay Volume Comparison: 1100/1200 Series Binary Pump vs. 1200 Series Binary Pump SLBinary Pump vs. 1200 Series Binary Pump SL

Binary pump SL (pressure range up to 600 bar):y p p (p g p )Standard delay volume configuration: 600-800μL (incl. damper and mixer)

Low delay volume configuration: 120μL (virtual damper)y g μ ( p )

Damper volume: 80-280μl

Binary pump (pressure range up to 400 bar):Standard delay volume configuration: 600-900μL (incl. damper and mixer)Standard delay volume configuration: 600 900μL (incl. damper and mixer)

Reduced delay volume configuration: ~200μL (damper needed)

Damper volume: 180μl + 1μl per bar


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All scaling calculations to transfer methods to RRHT, 1.8um particles are done using the Agilent Method Translator


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Features of the Agilent Method TranslatorBasic mode with certain pre-set parameters:as c ode ce a p e se pa a e e s

1.3.

2. 6.

4

5.

Enter the parameters of your existing method and the parameters of the

4.


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p y g pdesired column you would like to convert to.

Features of the Agilent Method TranslatorAdvanced mode – all calculation parameters in your hands:d a ced ode a ca cu a o pa a e e s you a ds

1 2.1.

[mL] [mL]

3.

4.

More to enter but much more information returned

3


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More to enter but much more information returned

Analysis of impurities of an active pharmaceutical ingredient by Does it work? - Example

conventional HPLC (4.6mmID x 250mm, 5.0µm):

mAU HN

CH3CH3

30

40 OH

OCH3

Main Compound

20

30

OH

HN

CH3CH3

OCHN

CH3CH3CHCHN

33

H

NCH3CH3

H

10

OCH3Impurity A

O CH3

Br

Bromanisole

OCH3

Impurity B

OCH3

Impurity C

OH

OH

Impurity D

0


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min0 2.5 5 7.5 10 12.5 15 17.5 20

Does it work?Converting to a 4.6 x 100 mm, RRHT column:


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mAU Conventional HPLC mAU35

Does it work? YES

30

40

25

30

35mAU

20

20

30

15

20

10

15

10

5

10

5

4 6 mm ID x 250 mm 5 0µm Zorbax SB C18

min0 2.5 5 7.5 10 12.5 15 17.5 20

0

4 6 mm ID x 100 mm 1 8µm Zorbax SB C18

min0 1 2 3 4 5 6 7 8

0

min0 0.5 1 1.5 2 2.5 3 3.5 4 4.5

0

4 6 mm ID x 100 mm 1 8µm Zorbax SB C184.6 mm ID x 250 mm, 5.0µm Zorbax SB C18

0.00 min 5% B20.00 min 90% B23.00 min 90% B23 01 min 5% B

4.6 mm ID x 100 mm, 1.8µm Zorbax SB C18


Simple Conversion4.6 mm ID x 100 mm, 1.8µm Zorbax SB C18


Speed Optimized


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23.01 min 5% B30.00 min 5% B23.01 min 5% B9.20 min 5% B4.99 min 5% B6.5 min 5% B

Advanced Mode: Select worst case viscosity for ACN/water at 40ºCACN/water at 40 C

0.75 cp


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Advanced Mode, Simple Conversion


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Advanced Mode, Resolution Optimized


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Using RRHT and other Low Volume HPLC Columns Effectively on Agilent 1200 and 1100 HPLCsEffectively on Agilent 1200 and 1100 HPLCs

• Use data acquisition rate of 0.1 sec• Use DAD SL for 80 Hz data acquisition• Short lengths of 0.12 mm i.d. tubing or smaller (watch pressure)• Thermostated column compartment plumbed through 3 μL sideThermostated column compartment plumbed through 3 μL side• For 2.1 mm id columns at elevated temps, use low vol. heat

exchangersF di t 80 L ( / 5022 2165) i d i j t• For gradients - 80 μL (p/n 5022-2165) or no mixer and injector bypass (not relevant for quaternary systems)

• Recommend micro and well plate autosamplers (ADVR “on”)• Otherwise, use injector program to reduce delay volume


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1100 System Configuration for Ultra-fast LC Recommendations for System Setup and Connecting Capillaries

Replace standard mixer of Binary Pump with 80 μL filter (p/n 5064-8273) to reduce delay volumneUse low volume, 3ul heat exchanger of TCC G1316A to th t t l t

1100 Binary Pump (G1312A) thermostate eluent

For 4.6 and 3mm columns use shortest possible 0.17mm ID connecting capillaries

Note: In ultra-fast applications the typical flow rate range using 4.6 and 3mm ID columns is 1-5 ml/min. At such higher flow

Pump (G1312A)

1100 WPS(G1367A)

4.6 and 3mm ID columns is 1 5 ml/min. At such higher flow rates the larger delay volume of 0.17mm ID capillaries doesn’t have a measurable negative impact on chromatographic performance.

For 2.1 and 1mm columns use shortest possible 0.12 or 0 1mm ID capillaries

3 μL heat exchanger

1100 TCC(G1316A)

0.1mm ID capillariesNote: In ultra-fast application the typical flow rate range using 2.1 and 1 mm ID columns is between 0.1-1 ml/min. At these lower flow rates smaller ID connecting capillaries should be used to minimize system delay volume and extra column peak dispersion/band broadening

4.6mm ID, 1.8umRRHT Column

dispersion/band broadening. Inlet tubing of the flow cell should be directly connected to the column.

Note: If this is not possible an appropriate low-volume connection should be used (capillary of small ID, i.e. 0.12 mm

0 17 d ZDV i )

Waste

1100 DAD SL(G1315C)


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or 0.17mm and ZDV-union).

Optimizing Gradient Separations With 1.8 um RRHT Columns: 10 X Faster Analysis

RRHT SB-C182.1 x 50mm, 1.8um

Conditions: Column: SB-C18, Dimensions listed below, Gradient: 10 – 90% ACN/25mM H3PO4, Gradient time: tG, as notedCPAH’s = Chlorphenoxyacid herbicides – environmental sample

i0 25 0 5 0 75 1 1 25 1 5 1 75 2

Temp: 50°CFlow: 1 mL/minGradient (tG): 2.4 min

C.

min0.25 0.5 0.75 1 1.25 1.5 1.75 2 2.25Rapid Resolution SB-C183.0 x 150mm, 3.5umTemp: 25°CFlow: 1.0 mL/minGradient (tG) : 18 min

B. Key Parameters• Particle size

Fl R t( G)

SB-C184 6 x 250mm 5um

0 2 4 6 8 10 12

A

• Flow Rate• Gradient Time• Column Length• Column ID• Temperature

min5 10 15 20 25

4.6 x 250mm, 5umTemp: 25°CFlow: 1mL/minGradient (tG): 30 min

A.p

Rsoptimized


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min5 10 15 20 25Sample: CPAH= Chlorophenoxy herbicides : Picloram, Chloramben, Dicamba, Bentazon, 2,4-D, Dichlorprop, 2,4,5-TP, Acifluorfen.

Translation to 3 0 x 150 mm 3 5 um 18 min gradient


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Translation to 3.0 x 150 mm, 3.5 um 18 min gradient

3.0 x 150 mm, 3.5 um, Resolution Optimized


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Scaling Gradients from 4.6 mm I.D. Columns to Solvent Saver Plus Column-Organic Acids

mAU

40506070 4.6 x 250 mm SB-C18, 5-um

57 mL solvent used25 uL std injection

1.5-mL/min; tg= 38 min

1

25 7

60

0102030

0min5 10 15 20 25 30 35

mAU70

4.6 x 150 mm SB-C18, 3.5-um 33 L l t d

3 45

6 8

10

010

20

3040

504.6 x 150 mm SB C18, 3.5 um 33 mL solvent used

15 uL std injection1.0-mL/min; tg= 33 min

20

40

60

30 35-10

mAUmin0 5 10 15 20 2580

3.0 x 100 mm SB-C18, 3.5-um10.5 mL solvent used

6 uL injection with INJ Program

min0 5 10 15 20 25 30 35-20

0

20

Analytes 1) gallic acid 3) protocatechuic acid 5) syringic acid 7) salicylic acid

0.5-mL/min; tg= 21min


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Analytes 1) gallic acid 3) protocatechuic acid2) hydrocaffeic acid 4) gentisic acid

5) syringic acid6) sinapinic acid

7) salicylic acid8) caffeic acid

Summary

• Method conversions are an opportunity to increase lab productivity significantly.

• The Agilent Method Translator is easy to use and can make your method translations to smaller columns much quicker and successful.

• Maintain resolution and avoid any change of selectivity y g y

• Proper choice of column size and efficiency,• Careful selection of method parameters.

• System optimization may be required to use smaller columns and/or smaller particle sizes (tubing, flow cell, delay volume, data rate)

• Increased operating pressure may result – ensure that system hasIncreased operating pressure may result ensure that system has adequate capacity for standard and increased pressure operation across the flow range of routine and optimized methods


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Appendix• Method translator link

– http://www.chem.agilent.com/en-us/products/instruments/lc/pages/gp60931 aspxus/products/instruments/lc/pages/gp60931.aspx

• Step-by-step Upgrade of 1100 to 1200 RRLC

– Pt 1 2 1mm ID columns Pub No 5989-6336EN– Pt. 1, 2.1mm ID columns, Pub. No. 5989-6336EN– Pt. 2, 4.6mm ID columns, Pub. No. 5989-6337EN

• Optimize Data Sampling Rate Pub No 5989-5810ENOptimize Data Sampling Rate, Pub. No. 5989 5810EN

• Agilent 1200 Series RRLC and RRLC/MS Optimization Guide, Pub. No. G1312-90301,

• Plug & Play Fast & Ultra-fast Separations Using 3.5um RR and 1.8um RRHT Columns, Pub. No. 5989-2908EN


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Wrap-up e-Seminar Questions

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