Analytical HPLC Method Validation of Amiloride and Its Pharmacokinetic Study in Humans

Abstract

This study was aimed to validate a reliable analytical method for the pharmacokinetic study of amiloride in human plasma by a high performance liquid chromatography (HPLC) system with UV detection. Triamterene was used as an internal standard. After extraction with ethylacetate, the supernatant was evaporated. Then, the residue was reconstituted and an aliquot was injected onto the HPLC system. Separation was performed on a Capcell Pak C 18 UG120 column (4.6 mm × 150 mm, 5 m particles) with a mobile phase of 12% acetonitrile containing 0.4% glacial acetic acid and UV detection at a wavelength of 360 nm. The intra-and inter-day precision expressed as the relative standard deviation was less than 15%. The flow rate of mobile phase was 1 mL/min and the retention time of amiloride and internal standard, triamterene, was found to be 3.06 and 8.80 min, respectively. The lower limit of quantification (LOQ) was 0.2 ng/mL of amiloride using 1 mL of plasma. The calibration curve was linear in the concentration range of 0.2-50 ng/mL (r 2 = 1 0000). The mean accuracy was 85.1-101.7%. The coefficient of variation (precision) in the intra-and interday validation was 2.1-12.5 and 2.4-13.7%, respectively. The pharmacokinetics of amiloride was evaluated after a single oral administration of 10 mg to healthy volunteers. The AUC 0-72hr , C max , T max , and T 1/2 were 267 7 ± 60 0 ng · hr/mL, 22 9 ± 5 4 ng/mL, 3 0 ± 0 8 hr, and 13 6 ± 2 4 hr, respectively. These results Correspondence: 2456 C. S. Myung et al.demonstrated that this method was highly feasible and reproducible for pharmacokinetic studies of amiloride in eight volunteers after oral administration (10 mg as amiloride HCl).

Stability Indicating Reverse Phase Ultra Performance Liquid Chromatography Method Development and Validation for Amiloride and Hydrochlorothiazide Tablet in the Presence of Degradation Products and Application to Green Analytical Principles

“…The substance under consideration is a solid of varying shades of yellow, ranging from light to dark, 229.63 g/mol as its molecular weight and its empirical formula is C 6 H 8 ClN 7 O. [1][2][3] Amiloride chemical structure is depicted in Figure 1.…”

Simultaneous quantification of amiloride and hydrochlorothiazide in human plasma by liquid chromatography–tandem mass spectrometry

“…Several methods have been reported to determine AMI as a single analyte in biological samples; however, most of them are very old (more than two decades) and were developed mainly using high performance liquid chromatography (HPLC). While very few recent methods are available for determination of AMI in human plasma [5] , [6] , detection or screening of AMI along with other diuretics in human urine especially for doping control has been discussed in several reports [7] , [8] , [9] . On the other hand, a number of methods have been described for the estimation of HCTZ either alone [10] , [11] , [12] or with other antihypertensive drugs in biological fluids [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] .…”