Simultaneous determination of irbesartan and hydrochlorothiazide in human plasma using HPLC coupled with tandem mass spectrometry: Application to bioequivalence studies

Faculty of Pharmacy, University of Jordan, Amman, Jordan.
Journal of pharmaceutical and biomedical analysis (Impact Factor: 2.98). 11/2009; 51(4):985-90. DOI: 10.1016/j.jpba.2009.10.023
Source: PubMed


A sensitive, specific and selective liquid chromatography/tandem mass spectrometric method has been developed and validated for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma. Plasma samples were prepared using protein precipitation with acetonitrile, the two analytes and the internal standard losartan were separated on a reverse phase C(18) column (50mmx4mm, 3microm) using water with 2.5% formic acid, methanol and acetonitrile (40:45:15, v/v/v (%)) as a mobile phase (flow rate of 0.70mL/min). Irbesartan and hydrochlorothiazide were ionized using ESI source in negative ion mode, prior to detection by multiple reaction monitoring (MRM) mode while monitoring at the following transitions: m/z 296-->269 and m/z 296-->205 for hydrochlorothiazide, 427-->175 for irbesartan. Linearity was demonstrated over the concentration range 0.06-6.00microg/mL for irbesartan and 1.00-112.00ng/mL for hydrochlorothiazide. The developed and validated method was successfully applied to a bioequivalence study of irbesartan (300mg) with hydrochlorothiazide (12.5mg) tablet in healthy volunteers (N=36).

Download full-text


Available from: Lara Tutunji, Mar 20, 2015
    • "High performance liquid chromatography with UV detection [15] and liquid chromatography-tandem mass spectrometry [16] have been used for the determination of losartan in human plasma. Hydrochlorothiazide has been determined alone or in combination with other drugs in biofluids using various methods among which are hyphenated techniques such as liquid chromatography mass spectrometry [17] [18] and liquid chromatography tandem mass spectrometry [19] [20]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The concept of personalized medicine is related to the development of new sensitive, precise and accurate analytical methods for therapeutic drug monitoring. In this article a rapid, sensitive and specific method was developed for the quantification of aliskiren, losartan, valsartan and hydrochlorothiazide in human plasma. Sample preparation was performed by protein precipitation with acetonitrile followed by filtration. All analytes and the internal standard (tiamulin) were separated by hydrophilic interaction liquid chromatography using an X-Bridge-HILIC analytical column (150.0 × 2.1 mm i.d., particle size 3.5 μm) under isocratic elution. The mobile phase was composed of a 10% 5 mM ammonium formate water solution pH 4.5, adjusted with formic acid, in acetonitrile and pumped at a flow rate of 0.25 mL min−1. The assay was linear over the concentration range of 5–500 ng mL−1 for all the analytes. Intermediate precision was less than 5.2% over the tested concentration ranges. The method is the first reported application of HILIC in the analysis antihypertensives in human plasma. With a small sample size (50 μL human plasma) and a run time less than 6.0 min for each sample the method can be used to support a wide range of clinical studies and therapeutic drug monitoring.
    No preview · Article · Sep 2015 · Journal of Chromatography B
  • Source
    • "Several methods have been reported for the analysis of the investigated AIIRA's, including high performance liquid chromatography [5] [6] [7] [8] [9], high pressure thin layer chromatography [10] [11] [12], capillary electrophoresis [13] [14] [15], voltammetric methods [16] [17] [18] [19] and spectrophotometric methods [20] [21] [22] [23] [24] [25]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: A simple, rapid, accurate and highly sensitive spectrofluorimetric method has been developed for determination of some angiotensin II receptor antagonists (AIIRA's), namely Losartan potassium (Los-K), Irbesartan (Irb), Valsartan (Val) and Candesartan cilexetil (Cand) in pure forms as well as in their pharmaceutical dosage forms. All the variables affecting the relative fluorescence intensity (RFI) were studied and optimized. Under the optimum conditions, linear relationships with good correlation coefficients (0.9982–0.9991) were obtained over the concentration range from 0.006 μg/mL to 1.7 μg/mL. Good accuracy and precision were successfully obtained for the analysis of tablets containing each drug alone or combined with hydrochlorothiazide (HCTZ) without interferences from the co-formulated HCTZ or the additives commonly present in tablets.
    Full-text · Article · Feb 2012 · Journal of Pharmaceutical Analysis
  • [Show abstract] [Hide abstract]
    ABSTRACT: Irbesartan, a diazaspiro angiotensin II blocker, is marketed in combination with Hydrochlorothiazide, which is a diuretic acting on distal convoluted tubule; for synergistic anti-hypertensive action. The present study deals with development and validation of a stability indicating HPTLC method for simultaneous estimation of Irbesartan and Hydrochlorothiazide using TLC plates precoated with Silica gel 60F254 and the mobile phase comprising Acetonitrile: Chloroform in the ratio of 5:6 v/v. Irbesartan and Hydrochlorothiazide were well resolved with Rf 0.27 ± 0.03 and 0.45 ± 0.03, respectively. Wavelength selected for the quantization was 270 nm. Inherent stability of these drugs was studied by exposing both drugs to various stress conditions as per ICH guidelines viz. Dry heat, oxidative, photolysis (UV and cool white fluorescent light) and hydrolytic conditions under different pH values. Both the drugs were not degraded under dry heat and photolytic conditions, but showed degradation under hydrolytic condition. The degraded products of Irbesartan and hydrochlorothiazide were well resolved from the individual bulk drug response. The developed method is found to be simple, specific, precise and stability indicating. The specificity of the method was confirmed by peak purity profile of the resolved peaks.
    No preview · Article · Oct 2010
Show more