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Dotsikas Y Apostolou C Soumelas S Kolocouri F Ziaka A Kousoulos C Loukas YL 《Journal of AOAC International》2010,93(4):1129-1136
An automated high-throughput HPLC/MS/MS method was developed for the quantitative determination of pantoprazole in human plasma. Only 100 microL plasma was placed in 2.2 mL 96 deep-well plates, and both pantoprazole and omeprazole (IS) were extracted from human plasma by liquid-liquid extraction, using diethyl ether-dichloromethane (70:30, v/v) as the organic solvent. Robotic liquid-handling workstations were used for all liquid transfer and solution preparation steps and resulted in a short sample preparation time. After vortexing, centrifugation, and freezing, the supernatant organic solvent was evaporated and reconstituted in a small volume of reconstitution solution. Sample analysis was performed by utilizing the combination of RP-HPLC/MS/MS, with positive-ion electrospray ionization and multiple reaction monitoring detection. The chromatographic run time was set at 1.8 min with a flow rate of 0.6 mL/min on a Nucleosil octylsilyl (C8) analytical column. The method was proven to be sensitive, specific, accurate, and precise for the determination of pantoprazole in human plasma. The method was applied to a bioequivalence study after per os administration of a 40 mg pantoprazole gastric retentive tablet. 相似文献
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Kolocouri F Dotsikas Y Apostolou C Kousoulos C Soumelas GS Loukas YL 《Journal of AOAC International》2011,94(3):758-764
An HPLC/MS/MS method characterized by complete automation and high throughput was developed for the determination of cilazapril and its active metabolite cilazaprilat in human plasma. All sample preparation and analysis steps were performed by using 2.2 mL 96 deep-well plates, while robotic liquid handling workstations were utilized for all liquid transfer steps, including liquid-liquid extraction. The whole procedure was very fast compared to a manual procedure with vials and no automation. The method also had a very short chromatographic run time of 1.5 min. Sample analysis was performed by RP-HPLC/MS/MS with positive electrospray ionization using multiple reaction monitoring. The calibration curve was linear in the range of 0.500-300 and 0.250-150 ng/mL for cilazapril and cilazaprilat, respectively. The proposed method was fully validated and proved to be selective, accurate, precise, reproducible, and suitable for the determination of cilazapril and cilazaprilat in human plasma. Therefore, it was applied to a bioequivalence study after per os administration of 2.5 mg tablet formulations of cilazapril. 相似文献
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Filomila Kolocouri Yannis Dotsikas Yannis L. Loukas 《Analytical and bioanalytical chemistry》2010,398(3):1339-1347
Dried plasma spots were employed as an alternative sample collection technique for the quantitative determination of gabapentin
in human plasma, using an automated liquid chromatography tandem mass spectrometry method. After the methanolic extraction
of single plasma, 1/8-in. disks which contained only 1.98 μL plasma volume, were placed in 96-well format plates, then gabapentin
and its internal standard, 4-aminocyclohexanecarboxylic acid, were derivatized with n-butanol/HCl (3 M) and detected as butyl esters. The assay exhibited excellent linearity over the concentration range of 40.0–10.0 × 103 ng/mL, which is suitable for the determination of gabapentin after per os administration of a single tablet. Variations in
intra- and inter-assay accuracy and precision were within internationally accepted criteria. Homogeneity of plasma spots was
proven, whilst butyl ester stability for both analytes was estimated and found very satisfactory. The quantitative analysis
of Gabapentin with dried plasma spot specimens seems to be a prominent and advantageous technique, especially when applied
to pharmacokinetic studies, where plasma sampling procedure becomes rapid and required plasma volumes negligible. 相似文献
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Kolocouri F Dotsikas Y Apostolou C Kousoulos C Loukas YL 《Analytical and bioanalytical chemistry》2007,387(2):593-601
An automated, sensitive and high-throughput liquid chromatographic/electrospray tandem mass spectrometric (LC–MS/MS) assay
was developed for the simultaneous determination of losartan (LOS), its major circulating metabolite EXP-3174 and hydrochlorothiazide
(HCTZ) in human plasma. LOS and HCTZ coexist in the same drug formulation, and this is the first method that enables the simultaneous
determination of both drugs along with the active metabolite of LOS. Since these drugs have different physicochemical properties,
the employment of a liquid–liquid extraction (LLE) protocol was precluded. A fully automated solid-phase extraction (SPE)
protocol, based on 96-well format plates, was used to isolate these compounds and furosemide (internal standard, IS) from
plasma. Washing and elution steps were amended accordingly in order to minimize any matrix effect from components of the plasma
without reducing the elution of the molecules of interest. The compounds were eluted from a C18 column and detected with an
API 3000 triple-quadrupole mass spectrometer using negative electrospray ionization and multiple reaction monitoring (MRM).
The assay was linear over the range 1.00–400 ng/mL for LOS and EXP-3174 and 0.500–200 ng/mL for HCTZ, respectively, when 200 μl
of plasma was used in the extraction. The overall intra- and interassay variations were within acceptance limits. The analysis
time for each sample was 4 min, and more than 300 samples could be analyzed in one day by running the system overnight. The
assay was simple, highly sensitive, selective, precise, fast, and it enables the reliable determination of LOS, EXP-3174 and
HCTZ in pharmacokinetic or bioequivalence studies after per os administration of a single tablet containing both drugs. 相似文献
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