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USE OF CAPILLARY ELECTROPHORESIS

FOR THE QUALITY CONTROL OF PHARMACEUTICAL

FORMULATIONS PRODUCED IN HOSPITAL PHARMACY

CEPharm, Boston, October 12th 2009

Susanne NussbaumerQuality control laboratory

Pharmacy of Geneva University Hospitals

Switzerland

Europe

SwitzerlandGeneva

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HUG HUG –– Geneva University Hospitals Geneva University Hospitals -- PharmacyPharmacy

2000 beds50’000 hospitalizations/year

50 employees

Pharmaceutical assistanceDistribution Production

Quality control laboratoryQuality control laboratory3 persons

~ 200 m2

> 20’000 analyses/year

Geneva University HospitalsGeneva University Hospitals

PharmacyPharmacy

Missions of Quality Control Laboratory HUGMissions of Quality Control Laboratory HUG

Prescription Production Quality control

Patient administration

Activities

Routine analysis Stability studies Incompatibility tests

Density, refractive index, melting pointboiling point, IR spectra, UV spectra…

End-products analysis

Cleanrooms/operators control

New formulation

Development, validation and application of analytical methods

Physicochemical incompatibilitiesbetween drugs

Analysis

Physicochemical analysis Microbiological analysis

Quantification of active drugs

Sterility testEndotoxine determination

Surface and area control

Raw material analysis

pH, osmolarity, particulate matteridentification of drugs/excipients…

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Production of Drugs at the Hospital PharmacyProduction of Drugs at the Hospital Pharmacy

Not available on the market

Available, but not in an appropriate formDosage (ex. pediatrics)Risk of error (ex. dilution)Risk of microbiological contamination (ex. intraophtalmics)Toxicity (ex. cytotoxics)

Clinical research

Reasons of production

Types of FormulationsInjectablesOphthalmicsOral, topic solutions SuspensionsSuppositoriesCapsules

Separation techniques used at HUG Ph. Separation techniques used at HUG Ph.

Illustrated by the recommendation of international pharmacopeias

LC-UVCost of column 1 column = 1 compound

High solvent consumption

High efficiency

Wide range of compounds

Different detectors

Rapid method development

Low cost of capillaries

Low solvent consumption CZE

MEEKC

NACE

MEKC CIEF

CITP

CE-UVCE-C4D

Before 2004Before 2004

After 2004After 2004

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codeine (Cod)oxybuprocaine (O)lidocaine (L), morphine (M) cocaine (Coc) procaine (IS)

min1 2 3

mAU

0

20

40

60

IST

E

PIS ISIS

K

IS

L

Coc M

O

Cod

IS

IS

ISIS

1 2 min

mAU

0

20

40

60

tetracaine (T)ephedrine (E)ketamine (K) phenylephrine (P)lidocaine (IS)

Experimental conditionsBGE: Tris-phosphate 50 mM, pH 2.5Injection: 5 s 20 mbarVoltage: 30 kVUV: 200 nm

CECE--UV: ShortUV: Short--end injection, aqueous BGEend injection, aqueous BGE

S. Fleury-Souverain, L. Vernez, C. Weber, P. Bonnabry. European Journal of Hospital Pharmacy. 15 (2009) 53-60.

Theoretical concentration

Morphine10 mg/mL

Trueness Repeatability (CV)

Intermediate precision (CV)

100.0% 0.8% 1.0%99.7% 0.9% 1.3%100.3% 1.0% 1.4%

Oxybu-procaine10 mg/mL

1.4% 1.5%1.0% 1.2%0.8% 1.0%

Tetracaine50mg/mL

0.8% 1.0%0.6% 0.6%0.4% 0.5%

Ketamine50 mg/mL

80%100%120%

80%100%120%

80%100%120%

80%100%120%

100.4%100.2%100.7%

99.7%100.5%99.7%

1.7% 2.0%2.7% 2.7%1.4% 2.4%

99.9%100.8%99.5%Ex

ampl

esof

val

idat

ion

resu

lts

Analyses of 5 pharmaceutical formulations by Analyses of 5 pharmaceutical formulations by CZE methods and CZE methods and ““oldold”” analytical methods.analytical methods.

106 % (LC)103 % (method 3)Homatropine

105% (LC)103% (method 2)Isoprenaline

104% (LC)107% (method 1)Ephedrine

88% (LC)87% (method 1)Phenylephrine

101% (single UV)103% (method 1)Cocaine collyre

Old analytical methodCZE methodsPharmaceutical formulations

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scopolamine (S)atropine (At) isoprenaline (I)adrenaline (Ad)procaine (internal standard (IS))

I

1 2

At

ISS

IS

IS

IS

Ad

min1 2

mAU

0

20

40

HydroHydro--organic BGEorganic BGE

Experimental conditions:BGE: Tris-phosphate 100 mM, pH 2.5 –

acetonitrile (80:20)Injection: 5 s 20 mbarVoltage: 30 kVUV: 200 nm

NonNon--aqueous BGEaqueous BGE

Experimental conditions:BGE: ammonium acetate 25 mM, acetic acid 1M

in MeOH-ACN (10:90, v/v)Injection: 10 s 40 mbarVoltage: 30 kV UV: 200 nm

homatropine ophthalmic solutionweak ophthalmic injection(homatropine (H), phenylephrine (P))procaine (internal standard (IS))

min1 2 3 4

mAU

40

min1 2 3 40

10

20

30

min1 2 3 4

IS

H

P

IS

H

S. Fleury-Souverain, L. Vernez, C. Weber, P. Bonnabry. European Journal of Hospital Pharmacy. 15 (2009) 53-60.

Stability Study: Cefuroxime 10 mg/mL 0.5mLStability Study: Cefuroxime 10 mg/mL 0.5mL

min2 4 6

mAU

0

10

20

30

40Ibuprofen (IS)

Cefuroxime

Degradationproducts

Experimental conditions:BGE: Phosphate buffer 20 mM pH 7.2Injection: 10 s 40 mbarVoltage: 30 kV UV: 200 nm

Validation results

Theoretical concentration

Trueness Repeatability (CV)

Intermediate precision

100.7% 1.3% 1.4%100.9% 1.3% 1.6%99.0% 1.5% 1.6%

80%100%120%

Stability-indicating method

Separation with degradation products

Satisfactory quantitative performances+Results of stability study

Syringes of cefuroxime at 10 mg/mL can be stored 4 months at –18°C without less of potency. After unfreezing, they must be used immediately (fast increase of degradation products at ambient temperature)

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Experimental conditions

BGE : 100 mM Tris-acetate at pH 4.2 acetonitrile (90:10, v/v)

Injection : 40 mbar 10sVoltage : 30 kVCapillary : 50 µm i.d., 375 µm o.d.

tot. length: 64.5 cm, eff. length: 50 cm min 1 2 3

mV

0

20

40

60 K (IS)SUX

Na

Choline

Validation

CECE--CC44D D –– suxamethoniumsuxamethonium

Suxamethonium (succinylcholine)

Stability study- ready-to-use solution - Succinolin, Lysthenon

S. Nussbaumer, S. Fleury-Souverain, S. Rudaz, P. Bonnabry, J.L. Veuthey. Journal of pharmaceutical and biomedical analysis. 49 (2009) 333-337.

Theoretical concentration

suxamethonium10 mg/mL

Trueness Repeatability (CV)

Intermediate precision (CV)

98.8% 1.1% 1.2%100.2% 1.3% 1.3%101.1% 0.6% 1.6%

80%100%120%

BGE : 100 mM Tris-acetate pH 4.5 : acetonitrile (80:20, v/v)

Injection : 40 mbar 10sVoltage : 30 kVCapillary : 50 µm i.d., 375 µm o.d.

total length: 64.5 cm, effective length: 50 cmC4D: output frequency: 150 kHz, output voltage: 40 Vpp

K

CaNaMg

Li(IS)

4 min2 3

mV

350

320

Experimental conditions

Quality control of total parenteral nutrition

CECE--CC44D: NaD: Na++, K, K++, Mg, Mg2+2+, Ca, Ca2+2+

S. Nussbaumer, S. Fleury-Souverain, L. Bouchoud-Bertholet, S. Rudaz, P. Bonnabry, J.L. Veuthey. Journal of pharmaceutical and biomedical analysis. Submitted to publication.

Validation

Theoretical Conc. [mM]

potassium

TruenessRepeatability

(CV)Intermediate precision (CV)

1 100.6% 1.0% 1.3%2 101.8% 1.2% 1.4%4 101.6% 1.1% 1.1%

sodium100.9% 1.2% 1.5%100.9% 1.1% 1.5%99.7% 0.9% 1.2%

calcium100.5% 1.1% 1.1%100.4% 1.3% 1.8%99.0% 0.4% 1.1%

magnesium99.1% 1.0% 1.2%99.2% 0.8% 1.1%98.6% 0.8% 0.8%

12 4

0.51 2

0.51 2

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CE analysis at the Ph. HUGCE analysis at the Ph. HUG

CE-UV/DAD CE-C4D

inorganic cationsin parenteral

nutrition

suxamethonium

Aqueous BGE Hydro-organic BGE

Non-aqueous BGE

codeine oxybuprocainelidocainemorphine cocaine procaine (IS)

tetracaineephedrineketaminephenylephrinelidocaine (IS)

scopolamine atropine isoprenalineadrenaline procaine (IS)

homatropineopht. solution

weak ophthalmic injection (homatropin, phenylephrine)procaine (IS)

cefuroxime

capacitively coupled contactless conductivity detection

Na+, K+, Ca2+, Mg2+

Quantitative analyses achieved byQuantitative analyses achieved by LCLC andand CECE

7 euro30 euroCost(capillaries or columns, consumables and products)

ConformConformQuantitative performances (trueness 100 ± 2%, CV <3%)

0 - 5 mL≥ 100 mLOrganic solvent consumption

3 h5 hTime

CELCMean CriteriaQuantitative analyse of one formulation batch:

20

40

60

80

100

2005 2006 2007 2008 Year

CE acquisition

2000-2004

% of separation analyses performed

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ConclusionConclusion

Use of CE in quality control laboratories should be strongly encouraged !

20 formulations analyzed by CE successfully by the HUG pharmacy,for routine analysis and stability tests

Environment respect

Similar performances

CE analysis is an attractive alternative to LC for the following reasons:

Economical aspects

Dr. Sandrine Fleury-SouverainProf. Pascal Bonnabry

Dr. Serge RudazProf. Jean-Luc Veuthey

AcknowledgementAcknowledgement

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Thanks for your attention!Thanks for your attention!