Simultaneous determination of five diuretic drugs using quantitative analysis of multiple components by a single marker

Abstract

Background: Loop diuretics are commonly used in clinical practice to manage high fluid loads and to control fluid balance. In this paper, a novel quantitative analysis method for multiple components with a single marker (QAMS) was developed for the simultaneous determination of 5 diuretic drugs furosemide, torasemide, azosemide, etacrynic acid, and bumetanide, by HPLC. Qualitative analysis was performed using relative retention time and ultraviolet (UV) spectral similarity as the double indicator. The QAMS method was conducted with etacrynic acid as an internal reference substance. The quantities of the other four diuretics were calculated by using the relative correction factors for etacrynic acid. The quantities of the 5 diuretic drugs were also determined by the external standard method (ESM). Chromatographic separation was achieved on a Shimadzu HC-C₁₈ column (150 mm × 4.6 mm, 5 µm) using 50 mM potassium dihydrogen phosphate (pH adjusted to 4.0 with phosphoric acid) with acetonitrile (64:36, v/v) as the mobile phase at a flow rate of 1.0 mL/min and a column temperature of 30 ℃.

Results: Under these conditions, the 5 diuretic drugs were well separated, showing linear relationships within certain ranges. The quantitative results showed that there was no significant difference between the QAMS and ESM methods.

Conclusions: Overall, the HPLC-QAMS analytical scheme established in this study is a simple, efficient, economical, and accurate method for the quantitative evaluation of 5 diuretic drugs.

Keywords: Diuretic drug; HPLC; Method development; Multi-components detection with a single marker.

Fig. 1

Chemical structures of furosemide (a), torasemide (b), azosemide (c), bumetanide (d) and etacrynic acid (e)

Fig. 2

A typical chromatogram of standard drug mixture samples. Chromatographic conditions were acetonitrile-0.05 mol·L⁻¹ potassium dihydrogen phosphate (36: 64, v/v); the buffer pH was varied. a. pH 3.0; b. pH 3.5; c. pH 4.0; d. pH 4.5; e. pH 5.0

Fig. 3

A typical chromatogram of standard drug mixture samples. Chromatographic conditions were acetonitrile-0.05 mol·L⁻¹ potassium dihydrogen phosphate (phosphoric acid to adjust pH to 4.0); the acetonitrile content was varied. a. acetonitrile content 30; b. acetonitrile content 34; c. acetonitrile content 36; d. acetonitrile content 38; e. acetonitrile content 40

Fig. 4

UV spectra and 1st UV spectra of furosemide, torasemide, azosemide, bumetanide and etacrynic acid