Simultaneous analysis of losartan, its active metabolite, and hydrochlorothiazide in human plasma by a UPLC-MS/MS method

Abstract

Abstract:A selective and sensitive ultra performance liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of losartan (LOS), EXP-3174, which is an active metabolite LOS carboxylic acid, and hydrochlorothiazide (HCTZ) in human plasma. Solid-phase extraction was carried out on Oasis HLB cartridges with 100 µ L of plasma to give an extraction recovery in the range of 88.5%-102.5% for the three analytes.Chromatography on a BEH C18 column afforded baseline separation of all the analytes within 2.4 min using 1.0% formic acid in water and acetonitrile (15:85, v/v) as the mobile phase. Quantitation was performed with multiple reaction monitoring in the negative ionization mode. The response of the method was linear over a dynamic range of 0.5-500, 1.0-750, and 0.25-150 ng/mL for LOS, EXP-3174, and HCTZ, respectively. Extent of signal suppression/enhancement was examined through postcolumn infusion. The effect of matrix components was evaluated by postextraction spiking and calculation of the slope of calibration lines. The method was successfully applied to a bioequivalence study of 50 mg losartan and 12.5 mg hydrochlorothiazide tablet formulation in 65 healthy human subjects. Reproducibility of the method was shown by reanalysis of 213 incurred samples.

Fast and sensitive LC–MS/MS method for the simultaneous determination of lisinopril and hydrochlorothiazide in human plasma

“…Several reports have used positive ionization mode for LC–MS/MS analysis of LIS [12] , [13] , while HCTZ which has polar groups gives a good mass spectrometric response in the negative mode [26] , [28] . As LIS is a polyfunctional, ampholyte molecule containing two basic and two acidic moieties (p K a 10.75, 7.13, 3.13 and 1.63), full scan mass spectra was acquired for both the analytes in the positive and negative ionization modes for electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) by direct infusion of 100 ng/mL solution of the analytes.…”

“…For chromatographic separation of these drugs various combinations of acetonitrile/ methanol and acidic buffers (ammonium formate/ammonium acetate) in the pH range of 3.0–5.5 were attempted in order to obtain symmetrical peak shapes, suitable retention and adequate signal-to-noise ratio leading to lower limits of quantitation. It was observed that the mobile phase composition and pH played a major role in chromatographic separation of these drugs as LIS has acidic as well as basic groups while HCTZ has p K a of 7.9 and 9.2 [28] . Compared to methanol, acetonitrile helped in providing higher sensitivity and sharp peaks shapes.…”

“…Few other methods present simultaneous determination of LIS together with other histamine H 2 antagonists [14] and other antihypertensive medications [15] , [16] in pharmaceutical formulations and human serum. Similarly, numerous methods are reported for the estimation of HCTZ in human plasma alone [17] , [18] , [19] , [20] , [21] or in combination with other antihypertensive drugs like bisoprolol [22] , candesartan [23] , irbesartan [24] , [25] , amlodipine and valsartan [26] , aliskiren and amlodipine [27] , losartan and losartan acid [28] , ramipril [29] and telmisartan [30] . So far only few reports are available for the simultaneous determination of LIS and HCTZ in biological samples [31] , [32] .…”

Bioequivalence study between a fixed‐dose single‐pill formulation of nebivolol plus hydrochlorothiazide and separate formulations in healthy subjects using high‐performance liquid chromatography coupled to tandem mass spectrometry

“…The run time ranged from 2.0 min (Shah et al, ) to 4.0 min (Kolocouri et al, ). The vast majority of methods employed ESI and the LOQ ranged from 0.25 ng/mL (Shah et al, ) to 8.0 ng/mL (Patel et al, ).…”

Simultaneous analysis of allopurinol and oxypurinol using a validated liquid chromatography–tandem mass spectrometry method in human plasma

“…The method was validated for system suitability, selectivity, carryover, sensitivity, linearity, accuracy and precision, recovery, matrix effect, stability, ruggedness and dilution reliability as per the current regulatory requirements [16] . The experimental details of the parameters studied were similar to our previous work [17] and are briefly described in Supplementary material.…”