use of capillary electrophoresis for the quality … · 2020. 6. 15. · aqueous bge hydro-organic...
TRANSCRIPT
1
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
2
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…
3
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
4
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
5
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)
6
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
7
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
8
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
9
Thanks for your attention!Thanks for your attention!