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Simultaneous determination of amiloride and hydrochlorothiazide in human plasma by liquid chromatography/tandem mass spectrometry with positive/negative ion‐switching electrospray ionisation
Author(s) -
Song Min,
Hang Taijun,
Zhao Hua,
Wang Li,
Ge Ping,
Ma Pengcheng
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3235
Subject(s) - chemistry , chromatography , selected reaction monitoring , mass spectrometry , electrospray ionization , hydrochlorothiazide , amiloride , tandem mass spectrometry , triple quadrupole mass spectrometer , elution , formic acid , rizatriptan , liquid chromatography–mass spectrometry , electrospray , analytical chemistry (journal) , medicine , biochemistry , receptor , organic chemistry , sumatriptan , blood pressure , radiology , agonist , sodium
A new method for simultaneous determination of amiloride and hydrochlorothiazide by liquid chromatography/electrospray tandem mass spectrometry (LC/MS/MS) operated in positive and negative ionization switching mode was developed and validated. Protein precipitation with acetonitrile was selected for sample preparation. The analytes were separated on a Phenomenex Curosil‐PFP (250 × 4.6 mm, 5 µm) column by a gradient elution with a mobile phase consisting of 0.15% formic acid solution containing 0.23% ammonium acetate and methanol pumped at a flow rate of 1.0 mL·min −1 . Rizatriptan was used as the internal standard (IS) for quantification. The determination was carried out on a Waters Quattro‐micro triple‐quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode using the following transitions monitored simultaneously: positive m/z 230 → 171 for amiloride, m/z 270 → 158 for rizatriptan, and negative m/z 296 → 205 for hydrochlorothiazide. The lower limits of quantification (LLOQs) were 0.1 and 1.0 ng·mL −1 for amiloride and hydrochlorothiazide, respectively, which were lower than other published methods by using ultraviolet (UV), fluorimetric or mass spectrometric detection. The intra‐ and inter‐day precision and accuracy were studied at three different concentration levels and were always better than 15% (n = 5). This simple and robust LC/MS/MS method was successfully applied to the pharmacokinetic study of compound amiloride and hydrochlorothiazide tablets in healthy male Chinese volunteers. Copyright © 2007 John Wiley & Sons, Ltd.