Simultaneous densitometric analysis of amlodipine, hydrochlorothiazide, lisinopril, and valsartan by HPTLC in pharmaceutical formulations and human plasma

ABSTRACT

A simple, accurate, and precise high-performance thin layer chromatographic (HPTLC) method has been developed and validated for the simultaneous quantification of antihypertensive drugs, amlodipine (AML), hydrochlorothiazide (HCTZ), lisinopril (LIS), and valsartan (VAL) in their pharmaceutical formulations and human plasma. Separation of the drugs was performed on aluminum-backed layer of silica gel 60 F₂₅₄ using a mixture of methanol–dichloromethane–glacial acetic acid (9.0:1.0:0.1, v/v/v) as the mobile phase. Densitometric determination of the separated spots was done at 215 nm. The retention factors (R_f) obtained under the optimized conditions were 0.56, 0.75, 0.29, and 0.67 for AML, HCTZ, LIS, and VAL, respectively. Linearity of the method was established in the range of 200–1,500 ng/band for AML, 300–1,500 ng/band for HCTZ, 400–2,000 ng/band for LIS, and 1,000–7,000 ng/band for VAL. The limit of detection/limit of quantitation of the method found were 54.21/164.28, 77.27/234.15, 83.45/252.87, and 156.48/474.19 ng/band for AML, HCTZ, LIS, and VAL, respectively. To determine the drugs in spiked plasma samples, solid phase extraction was performed, which provided highly consistent and quantitative recovery for all four drugs. The method was satisfactorily applied for the analysis of different tablet formulations and proved to be specific and accurate for the quality control of these drugs.

GRAPHICAL ABSTRACT

KEYWORDS:

• Antihypertensive drugs
• high-performance thin layer chromatography
• human plasma
• pharmaceutical formulations
• uncertainty determination