Development of HPLC-DAD method for the quantification of pentostatin in the fruiting body of Cordyceps militaris

Abstract

Cordyceps militarisis an important medicinal mushroom in traditional medicine. It contains many bioactive compounds that have applications in medicine, such as cordycepin, adenosine... Recently, through genome analysis, C. militaris has been shown to have the ability to synthesise pentostatin. Pentostatin is an important active ingredient in the production of chemotherapy drugs used in cancer treatment. However, there have been few studies on the development of methods for quantifying pentostatin in C. militaris. In this study, a simple and sensitive high-performance liquid chromatography with diode-array detection (HPLC-DAD) method for the quantification of pentostatin in C. militaris was developed and validated. The chromatographic separations were obtained by an Agilent C₁₈ column (250x4.6 mm i.d., 5 µm particle size) using gradient elution with acetonitrile and water. The flow rate of 0.6 ml/min and detection wavelength of 280 nm were selected. The calibration curve displayed a good linear correlation (R²>0.99). The validation results displayed great accuracy, repeatability, linearity, and selectivity according to The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guideline.