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Bioanalytical method development and validation of simultaneous analysis of telmisartan and hydrochlorthiazide in human plasma using LC-MS/MS

Mathew George*, Lincy Joseph, Arpit Kumar Jain, Anju V.


A simple, sensitive, rapid and economic high performance thin layer chromatographic method and a mass spectroscopic assay method has been developed for the quantification of telmisartan and hydrochlorthiazide combination in human plasma. The internal standards and analytes were extracted from human plasma by solid-phase extraction with HLB Oasis1cc (30mg) catridges. The scanning and optimization for the samples are done using methanol: water (50:50). The samples were chromatographed using reverse phase chromatography with C-18 column of different manufacturers like Ascentis C18 (150×4. 6, 5µ) using the buffer system Acetonitrile: Buffer (80:20%v/v) which consist of 2±0. 1Mm ammonium format at a flow rate of 0. 7ml/min at a column oven temperature 35±10c. The internal standard used was hydrochlorthiazide13c1, d2 and telmisartand3. The extraction techniques include conditioning, loading, washing and elution, drying followed by reconstitution of the dried samples. The volume injected was 10µl with the retention time of 3-4 min for telmisartan, 1-2 min for hydrochlorthiazide and for the internal standards the retention time was 3-4 min for telmisartand3 and 1-2 min for hydrochlorthiazide c13d2. The rinsing solution was Acetonitrile: HPLC grade water in the ratio (50:50). The above developed method was validated using various parameters like selectivity and sensitivity, accuracy and precision, matrix effects, % recovery and various stability studies. The method was proved to be sensitive, accurate, precise and reproducible. The preparation showed high recovery for the quantitative determination of telmisartan and hydrochlorthiazide in human plasma.


Telmisartan; Hydrochlorthiazide; Solid phase extraction; Liquid chromatography –Mass spectroscopy; Bioanalytical validation.

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Ruth, R. W, J. C. William, D. I. John, et al., “Nonpeptide angiotensin II receptor antagonists: the next generation in antihypertensive therapy” J. Med. Chem., 39. 2 (1996): 625–656.

Yusuf, S, K. K. Teo, J. Pogue, et al., “Telmisartan, ramipril, or both in patients at high risk for vascular events” N. Engl. J. Med., 358. 15 (2008): 1547–1559.

Rane V. P, Sangshetti J et al., Simultaneous high performance liquid chromatographic determination of telmisartan and hydrochlorthiazide in pharmaceutical preparation” Journal of chromatographic Science. 46. 10 (2008):887-891.

Gilroy J. J, Dolan J. Wet al., “Column sensitivity in reversed phase liquid Chromatography-I, Type B –alkyl silica columns” Journal of chromatography A 1026. 1-2: (2003):757-778.

C. K, Hull, Penman A. D, Smith C. K et al., “Quantification of rosuvastatin in human plasma by automated solid phase extraction using tandem mass spectroscopic detection” Journal of chromatography. B 772. 2 (2002):219-28.

Waters HPLC Column and reverse phase mobile selection guidelines (2012):1-34.

Gerhards P, Sadjdi. S. “Rapid HPLC column and mobile phase screening for fast LCMS/MS development” (2004):1-3.

Scott E, VanBramer. “An introduction to mass spectrometry”. Widener University, Department of chemistry (1998):4-33.

Holffmann E. D “Tandem Mass Spectrometry: A primer”. Journal of mass spectrometry. 31 (1996): 129-137.

Ravi, B, Vasu, Jaswanth Kumar et al., “Simultaneous determination of telmisartan and amlodipine in human plasma by LC–MS/MS and its application in a human pharmacokinetic study”. Journal of pharmaceutical Analysis2. 5 (2012):319-326.

Salama, Ismail. “Simultaneous HPLC–UV analysis of telmisartan and hydrochlorothiazide in human plasma”. Bulletin of faculty of pharmacy;49. 1 (2011):19-24.

Rote, R, Ambadas, Poonam. R. Sonavane. “Development and validation of bioanalytical method for determination of telmisartan and hydrochlorthiazideusing HPTLC in human plasma". American J. Analytical Chemistry 3 (2012):774-778.

Ganesh, Shankar, Pragney Deme. “Simultaneous determination of amlodipine, Valsartan and HCTZ by LC-ESI-MS/MS and it’s application in pharmacokinetic in rats” J Pharm Analysis, 4. 6 (2014): 399-406.

Raghunadha Reddy Seelam, Raghu Sarath Chandiran. I. “Development and validation of high performance LC-TMS method for simultaneous quantification of telmisartan in human plasma”. International Journal of Pharmaceutical Science and Drug Research. 2. 3 (2010): 188-192.

Patel, M, Jaydeep, Anjali. P. Dinghani. “Development and validation of bioanalytical HPLC method for estimation of telmisartan in rat plasma. Application of pharmacokinetic studies”. 11. 2 (2012): 121-127.

Lan, KeXuehuajiang, Yunxia Li, Ling Wang et al., “Quantitative determination of rosuvastatin in human plasma by ion pair liquid-liquid extraction using liquid chromatography with electrospray ionization tandem mass spectrometry” Journal of pharmaceutical and biomedical analysis 44. 2 (2007): 540-546.

Shah V. P, Midha K. K, Findlay J. W. A et al., “Analytical method validation: Bioavailability, bioequivalence and pharmacokinetic studies”. Pharm Res 9 (1992): 588- 592.

S, Bansal, Destefano. “A Key elements of bioanalytical method validation for small molecules” AAPS Journal 9. 1 (2007) E109-114.

R. R, Kalakuntla, Santhosh. K “Bioanalytical Method Validation: Aquality assurance auditor view point” J. Pharm Sci and research. 3 (2009):1-10.

B. ChenLiang. Y et al., “Development and validation of liquid chromatography-mass spectrometry method for the determination of telmisartan in human plasma”. Analytica Chimica Acta 540 (2005):367-373.

Yana. T, Hao Li et al., Liquid chromatographic tandem mass spectroscopic method for the simultaneous quantitation of telmisartan and hydrochlorthiazide in human plasma. J. Pharm Bio MED Analysis 48; 2008; pg no:1225-1229.

H, Zhang, Yunyun Y et al., “Rapid determination of telmisartan in human plasma by HPLC using a monolithic column with flouresecence detection and its applications to a bioequivalence study”. Journal of chromatographyB 877. 29 (2009):3729-3733.

Vijayabharathi, D, Kishore Kumar Hothaet al., “LC–MS/MS method for simultaneous estimation of candesartan and hydrochlorothiazide in human plasma and its use in clinical pharmacokinetics”. 4. 10 (2009):1195-1204.

Kumar, Suresh, James. D. Terish, N. Ramesh “Estimation of Telmisartan in Human Plasma by Reversed Phase Liquid Chromatography Coupled with Tandem Mass Spectrometry-A Bioequivalence Study Application” Der Pharmacia Lettre 3.4 (2011): 289-298.



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