title: A single, selective and simple validated method for simultaneous estimation of amiloride and hydrochlorothiazide in human plasma by liquid chromatography-tandem mass spectrometry.
abstract: A single, simple and selective method for simultaneous estimation of amiloride and hydrochlorothiazide in human plasma was validated using triamterine and hydrochlorothiazide (13)C,d2 as internal standard. The compounds were separated on a reverse-phase column with an isocratic mobile phase consisting of 2 mm 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 (13)C,d2 in negative ion mode. Linearity in plasma was observed over the concentration range 0.1-10 ng/mL for amiloride and 5.0-500.0 ng/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.
authors.affiliation: School of Chemical Sciences, Swami Ramanand Teerth Marathwada University, Nanded, India. jangid_arvind@rediffmail.com
authors.forename: Arvind G
authors.initials: AG
authors.lastname: Jangid
authors.forename: Rajesh H
authors.initials: RH
authors.lastname: Tale
authors.forename: Vikas V
authors.initials: VV
authors.lastname: Vaidya
mesh_terms: D000584:Amiloride / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D006852:Hydrochlorothiazide / Q000097:blood*; D016018:Least-Squares Analysis; D057230:Limit of Detection; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006852:Hydrochlorothiazide; D000584:Amiloride