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Jangid A.G.,Swami Ramanand Teerth Marathwada University | Jangid A.G.,Accutest Research Laboratories I Pvt. Ltd. | Tale R.H.,Swami Ramanand Teerth Marathwada University | Vaidya V.V.,Swami Ramanand Teerth Marathwada University
Biomedical Chromatography | Year: 2012

A single, simple and selective method for simultaneous estimation of amiloride and hydrochlorothiazide in human plasma was validated using triamterine and hydrochlorothiazide 13C,d2 as internal standard. The compounds were separated on a reverse-phase column with an isocratic mobile phase consisting of 2mm ammonium acetate pH 3.0 and acetonitrile (30:70, v/v) and detected by tandem mass spectrometry with positive/negative ion mode. The analytes and internal standards were extracted from plasma using simple solid phase extraction. The ion transitions recorded in multiple reaction monitoring mode were m/z 230.1→116.0 for amiloride, m/z 254.1→237.1 for internal standard, triamterine in positive mode and m/z 296.1→204.9 for hydrochlorothiazide, m/z 299.2→205.8 for internal standard, hydrochlorothiazide 13C,d2 in negative ion mode. Linearity in plasma was observed over the concentration range 0.1-10ng/mL for amiloride and 5.0-500.0ng/mL for hydrochlorothiazide. The mean recovery was 41.1 and 81.5% for amiloride and hydrochlorothiazide respectively. The coefficient of variation of the assay was less than 11.2 and 5.2% for amiloride and hydrochlorothiazide, respectively, and the accuracy was 89.0-98.1 and 96.6-102.9% for amiloride and hydrochlorothiazide, respectively. The validated method can be applied to the pharmacokinetic study of amiloride and hydrochlorothiazide. Copyright © 2011 John Wiley & Sons, Ltd. Source


Pudage A.,University of Mumbai | Pudage A.,Accutest Research Laboratories I Pvt. Ltd. | Kamat S.,University of Mumbai
Biomedical Chromatography | Year: 2011

In this paper, we present a validated UPLC-MS/MS assay for determination of ramipril and ramiprilat from human plasma samples. The assay is capable of isolating phase II metabolites (acylglucornides) of ramipril from in vivo study samples which is otherwise not possible using conventional HPLC conditions. Both analytes were extracted from human plasma using solid-phase extraction technique. Chromatographic separation of analytes and their respective internal standards was carried out using an Acquity UPLC BEH C18 (2.1×100mm), 1.7μm column followed by mass spectrometric detection using an Waters Quattro Premier XE. The method was validated over the range 0.35-70.0ng/mL for ramipril and 1.0-40.0ng/mL for ramiprilat. © 2010 John Wiley & Sons, Ltd. Source


Shinde V.P.,Kadi Sarva Vishvavidyalaya | Shinde V.P.,Accutest Research Laboratories I Pvt. Ltd. | Pudage A.,Accutest Research Laboratories I Pvt. Ltd. | Jangid A.,Accutest Research Laboratories I Pvt. Ltd. | And 3 more authors.
Journal of Pharmaceutical Analysis | Year: 2013

A novel, fast, sensitive and robust method based on ultra-performance liquid chromatography coupled to atmospheric pressure electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) has been developed to separate two Tibolone stereoisomers i.e., 3α-Hydroxy Tibolone and 3β-Hydroxy Tibolone and to quantify 3α-Hydroxy Tibolone using p-toulenesulfonyl isocyanate (PTSI) as a derivatizing reagent in human plasma. 3α-Hydroxy Tibolone-13CD3 was used as an internal standard (IS). The analyte and IS were extracted from human plasma by liquid-liquid extraction using ethyl acetate. Extracted samples were analyzed by UPLC-ESI-MS/MS. Chromatography was performed using binary gradient on UPLC analytical column. A linear calibration curve over the range of 0.100-35.000 ng/mL was obtained and lower limit of quantification (LLOQ) was 0.100 ng/mL demonstrating acceptable accuracy and precision. This method was successfully applied to a pharmacokinetic study in order to compare a test Tibolone 2.5 mg formulation vs. a reference 2.5 mg Tibolone tablet formulation in 50 post-menopausal/surgical menopause female human volunteers under fasting conditions. It is concluded that test formulation of Tibolone is bioequivalent to reference formulation of Tibolone. © 2013 Xi'an Jiaotong University. Source


Jangid A.G.,Accutest Research Laboratories I Pvt. Ltd. | Jangid A.G.,Swami Ramanand Teerth Marathwada University | Pudage A.M.,Swami Ramanand Teerth Marathwada University | Joshi S.S.,Swami Ramanand Teerth Marathwada University | And 2 more authors.
Biomedical Chromatography | Year: 2010

A rapid, simple and specific method for estimation of anastrazole in human plasma was validated using letrozole as internal standard. The analyte and internal standard were extracted from plasma using simple solid-phase extraction. The compound were separated on a reverse-phase column with an isocratic mobile phase consisting of 0.1% formic acid in water and acetonitrile (12 : 88, v/v) and detected by tandem mass spectrometry in positive ion mode. The ion transitions recorded in multiple reaction monitoring mode were m/z 294.1 → 225.1 for anastrazole and m/z 286.1 → 217.1 for internal standard. Linearity in plasma was observed over the concentration range 0.3-30 ng/mL for anastrazole. The mean recovery for anastrazole was 83.7% with a lower limit of quantification of 0.3 ng/mL. The coefficient of variation of the assay was less than 6.8% and the accuracy was 96.1-102.2%. The validated method was applied to a bioequivalence study of 1 mg anastrazole tablet in healthy human volunteers. Copyright © 2009 John Wiley & Sons, Ltd. Source

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