Simultaneous determination of adenine, uridine and adenosine in cordyceps sinensis and its substitutes by LC/ESI-MS

Abstract

A simple, sensitive and reproducible high performance liquid chromatography-mass spectrometry coupled with electrospray ionization method for simultaneous separation and determination of adenine, adenosine and uridine was developed. The analytical column is a 2.0 mm × 150 mm Shimadzu VP-ODS column and volume fraction of the mobile phase is 86.5% water, 12.0% methanol and 1.5% formic acid. 2-chloroadenosine was used as internal standard. Selective ion monitoring mode and selective ion monitoring ions at ratio of mass to electric charge of 136 for adenine, 268 for adenosine and 267 for uridine were chosen for quantitative analysis of the three active components. The results show that the regression equations and linear range are Y=0.062X+0.005 and 2.0 – 140.0 µg·mL⁻¹ for adenine, Y=0.049X+0.004 and 4.0–115.0 µg · mL⁻¹ for uridine, Y=0.154X+0.014 and 1.0–125.0 µm · mL⁻¹ for adenosine. The limits of detection are 0.6 µg·mL⁻¹ for adenine, 1.0 µg·mL⁻¹ for uridine and 0.2 µg·mL⁻¹ for adenosine. The recoveries of the three constituents are from 96.6% to 103.2%.

Keywords

high performance liquid chromatography / electrospray ionization interface / mass spectrometry / cordyceps sinensis / nucleoside