Development of an HPLC method to analyze and prepare elsinochrome C and hypocrellin A in the submerged fermentation broth of Shiria sp. SUPER‐H168

A rapid and sensitive analytical method based on reverse‐phase high‐performance liquid chromatography was first developed to simultaneously determine elsinochrome C (EC) and hypocrellin A (HA) in the submerged fermentation. The mobile phase consisted of acetonitrile–water 60:40 (v/v) with a flow‐rate of 1 mL/min. The calibration curves were as follows: y  = 37,625 x  + 249,775 for EC, y  = 30,813 x  + 556,409 for HA and linear at the investigated concentration. The correlation coefficients ( R 2 ) were 0.9989 and 0.9998 respectively for EC and HA. The limits of detection and quantification were 175 and 585 µg/L for EC and 205 and 610 µg/L for HA. The precisions of concentration and retention times were less than 2.5 and 0.3%. The recovery of the method was greater than 95.0%. The methodology was applied to analyze simultaneously EC and HA concentrations in a submerged fermentation, and was adequate for analysis of biosynthesis of perylenequinones. The method was also amplified to separate and purify EC and HA using a semi‐preparative C 18 column. In addition, elsinochrome C was first identified in the submerged fermentation broth of Shiraia sp. SUPER‐H168. Copyright © 2011 John Wiley & Sons, Ltd.