Simple RP-HPLC method for determination of triple drug combination of valsartan, amlodipine and hydrochlorothiazide in human plasma

Abstract

Hypertension is emerging as one of the most significant health complications in recent years (1). There has been a marked increase in the use of combinations of antihypertensive drugs as well with complementary mechanisms of action, with the aim of reducing blood pressure levels more rapidly and improving treatment compliance. The 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 C 18 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 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 demostrated 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.

Application of an LC–MS/MS method for the analysis of amlodipine, valsartan and hydrochlorothiazide in polypill for a bioequivalence study

“…To the best of our knowledge, there are only four methods which describe their simultaneous analysis in pharmaceutical preparations [42] , [43] and human plasma [42] , [43] , [44] , [45] . The salient features of these methods are summarized in Table 1 .…”

Statistical Optimization of Extraction Process for the Quantification of Valsartan in Rabbit Plasma by a HPLC Method

“…HPLC with UV detectors are easy to handle. The HPLC methods with photodiode array (PDA) and UV detectors have also been reported for the extraction of valsartan from the human plasma [1,18,19] .…”

“…The flow rate and stationary phase used were 1.0 ml/min and Gemini C 18 (250×4.6 mm i.d., particle size 5 µm) column (Phenomenex ® , USA) protected with C 18 (4×2 mm i.d.) guard column, respectively [19] . However, these existing HPLC-UV and HPLC-PDA methods involved multiple steps of extraction and use of expensive gases (such as nitrogen) for evaporation of organic solvents.…”

Bioanalytical Method Development and Validation for Quantitative Estimation of Valsartan by LC-MS/MS in Human Plasma

“…Literature survey revealed that few analytical methods based on HPLC [7][8][9][10][11][12] are reported for the determination of valsartan in biological samples. Conventional HPLC methods have the limitations in chromatographic run time, specificity and sensitivity compared with LC-MS/MS methods.…”