Development of RP HPLC-PDA method for simultaneous quantitative analysis of Inoscavin A and Meshimakobnol A and application on some Phellinus mushroom species.

Phellinus igniarius, a medicinal mushroom containing many active ingredients with health benefits, can be applied in functional food. At present, the quantification of the main active ingredients from higher fungi (Ganoderma, Phellinus…) materials from different growing sources is a mandatory requirement to standardize the input resources of pharmaceutical and food production. Our study's aims are to perfect the RP HPLC-PDA method for quantitative analysis of Inoscavin A and Meshimakobnol A which are two main active ingredients present in Phellinus mushroom. In this analytical method, a C18-HPLC column and the mixture of methanol and formic acid solutions (pH = 2.2) are used to analyze and elute the active substances with the column activity parameters being the concentration gradient. This perfect method was tested for system suitability, repeatability, intermediate precision, recovery, and linear curve calibration to validate the method. After validation, the perfected RP HPLC-PDA method was applied to analyze eight samples of Phellinus and three samples of Ganoderma mushroom category. This method can be the basis for classifying between Phellinus and some other medicinal mushrooms.

HPLC quantitative analysis of protocatechuic acid contents in 11 Phellinus mushroom species collected in Thailand

Abstract Phellinus mushrooms have been traditionally used for various medicinal purposes. Protocatechuic acid, which was previously reported to be a component in some Phellinus mushrooms, has some pharmacological effects. This study aimed to validate a HPLC method for the quantitative analysis of the protocatechuic acid contents in the extracts from different Phellinus mushroom species collected in Thailand. HPLC was carried out using a C18 column and the gradient mobile phases of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Method validation was performed to assure the linearity, accuracy, precision, limit of detection and limit of quantitation of the analytical method. The linearity range of protocatechuic acid was 1 - 10 µg/ml. The average recovery was 104.16%. The method was shown to be precise with the RSD of repeatability and intermediate precision at less than 3%. The protocatechuic contents in 11 Phellinus mushrooms were in the range of less than 0.0099 - 0.4121 %w/w of the extract. The developed HPLC method was reliable and suitable for the quantitative analysis of protocatechuic acid content in Phellinus mushrooms.