Acta Pharm. 62 (2012) 45-58
Original research paper
Simple RP-HPLC method for determination of
triple drug combination of valsartan, amlodipine and hydrochlorothiazide in
human plasma
RITESH N. SHARMA and SHYAM SUNDER PANCHOLI
riteshn.sharma@gmail.com
1 S. K. Patel College of
Pharmaceutical Education and Research, Ganpat University, Mahesana-Gozaria
Highway, Kherva Dist. Mahesana-382711, Gujarat, India
2 Babria
Institute of Pharmacy, Varnama, Vadodara-391240, Gujarat, India
Accepted January 2, 2012
A simple RP-HPLC method for the quantification of valsartan (VAL), amlodipine (AML) and hydrochlorothiazide (HCT) in human plasma was developed and validated. VAL, AML and HCT were resolved using a Gemini C18 column and mobile phase gradient starting from 20 % acetonitrile and 80 % 10 mmol L-1 ammonium formate (V/V, pH 3.5 ± 0.2, by formic acid) to 70 % acetonitrile and 30 % 10 mmol L-1 ammonium formate, over 20 minutes with a flow rate of 1 mL min-1. The samples were purified by protein precipitation and extraction. Telmisartan was used as internal standard. The method was validated according to USFDA and EMEA guidelines with good reproducibility and linear responses R = 0.9985 (VAL), 0.9964 (AML), and 0.9971 (HCT). RSDs of intra- and inter-day precision ranged between 2.2-8.1 and 4.6-11.7 %, respectively, for all three drugs. Mean extraction recoveries of three QCs for the triple drug combination were 76.5 (VAL), 72.0 (AML) and 73.0 (HCT) % for human plasma. Although the LC-MS/MS method is more sensitive than HPLC, HPLC is still suitable for preliminary pharmacokinetic study. The experiments performed demonstrated that simultaneous determination of all components of the triple drug combination in human plasma can be done by this method. Proposed method can be also used for guidance to the LC-MS/MS method.
Keywords: valsartan,
amlodipine, hydrochlorothiazide, triple drug combination, plasma, RP-HPLC, preliminary pharmacokinetic