ABSTRACT
A specific liquid chromatography-mass spectrometric (LC-MS/MS) assay was developed and validated for the determination of lercanidipine, a dihydropyridine calcium channel blocker, in human plasma. Lercanidipine R-D3 was used as internal standard (IS). The drug was extracted from plasma using liquid-liquid extraction technique utilizing hexane: ethyl acetate as extraction solvent. The samples were analyzed using a prepacked Thermo Hypersil C(8) column and a mobile phase composed of a mixture of aqueous acetic acid and triethylamine in methanol. An ion trap mass spectrometer equipped with electrospray ionization (ESI) source operating in the positive ion mode was used to develop and validate the method. The method was proved to be sensitive and specific by testing six different human plasma batches. Linearity was established for the concentration ranges of 0.1-16 ng/ml with a regression factor of 0.9996. The lower limit of quantitation was identifiable and reproducible at 0.1 ng/ml with a precision of 7.2%.
Subject(s)
Calcium Channel Blockers/blood , Chromatography, High Pressure Liquid/methods , Dihydropyridines/blood , Spectrometry, Mass, Electrospray Ionization/methods , Humans , Reference Standards , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
A sensitive and specific liquid chromatography electrospray ionization ion-trap mass spectrometry (LC-ESI-IT-MS/MS) method has been developed and validated for the identification and quantitation of loratadine in human plasma. After the addition of the internal standard (IS), plasma samples were extracted using isooctane:isoamyl alcohol mixture. The compounds were separated on a prepacked Zorbax phenyl column using a mixture of acetonitrile, 0.20% formic acid as mobile phase. A Finnigan LCQ(DUO) ion-trap mass spectrometer connected to a Waters Alliance high performance liquid chromatography (HPLC) was used to develop and validate the method. The results were within the accepted criteria as stated in the FDA bioanalytical method validation guidance. The method was proved to be sensitive and specific by testing six different plasma batches. Linearity was established for the range of concentrations 0.10-10.0 ng/ml with a coefficient of determination (r(2)) of 0.9998. Accuracy for loratadine ranged from 105.00 to 109.50% at low, mid and high levels. The intra-day precision was better than 10.86%. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 0.10 ng/ml with a precision of 9.84%. The proposed method enables the unambiguous identification and quantitation of loratadine for pharmacokinetic, bioavailability or bioequivalence studies.
Subject(s)
Loratadine/blood , Spectrometry, Mass, Electrospray Ionization/methods , Chromatography, Liquid/methods , Humans , Loratadine/chemistryABSTRACT
A sensitive and specific high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS-MS) method has been developed at our center for the determination of glimepiride in human plasma. After the addition of the internal standard, plasma samples were extracted by liquid-liquid extraction technique using diethyl ether. The compounds were separated on a prepacked C18 column using a mixture of acetonitrile, methanol and ammonium acetate buffer as mobile phase. A Finnigan LCQDUO ion trap mass spectrometer connected to an Alliance Waters HPLC was used to develop and validate the method. The analytical method was validated according to the FDA bioanalytical method validation guidance. The results were within the accepted criteria as stated in the aforementioned guidance. The method was proved to be sensitive and specific by testing six different plasma batches. Linearity was established for the range of concentrations 5.0-500.0 ng/ml with a coefficient of determination (r2) of 0.9998. Accuracy for glimepiride ranged from 100.58 to 104.48% at low, mid and high levels. The intra-day precision was better than 12.24%. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 5.0 ng/ml with a precision of 7.96%. The proposed method enables the unambiguous identification and quantitation of glimepiride for pharmacokinetic, bioavailability or bioequivalence studies.
