Extraction, Screening and Mass Spectrometry Analysis of Active Triterpenoid Components in Ganoderma lucidum
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Abstract:
In this study, the optimal extraction conditions of the total triterpenes from Ganoderma lucidum were investigated, and the inhibitory effect of triterpenoids on lipoxygenase was evaluated. Single factor experiments and response surface method were used to analyze the effects of extraction time, extraction times, liquid-to-material ratio, and ethanol concentration on the yield of triterpenes from Ganoderma lucidum, and screen the optimal process parameters. High performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to analyze and identify the chemical constituents of the triterpenoid extracts from Ganoderma lucidum. Lipoxygenase was used as the biological target molecule, and ultrafiltration mass spectrometry was used to screen enzyme inhibitors and examine the inhibitory effect of the triterpenoids on lipoxygenase. The results revealed that the optimal process conditions were material-to-liquid ratio of 1:30, ethanol concentration of 75%, three extraction times, and extraction time of 1.5 h. Under these conditions, the extraction rate of total triterpenes was 1.04%; five types of lipoxygenase inhibitors were screened out from Ganoderma lucidum, which demonstrated a strong inhibitory effect on the activity of lipoxygenase at 1.0 U/mL. The order of the inhibitory effect was as follows: ganoderic acid B (52.64%) >ganoderic acid C2 (40.21%) >ganoderic acid D2 (31.25%) >ganoderic acid F (12.69%) >ganoderic acid A (10.82%). The experimental results showed that the extraction process parameters of red Ganoderma lucidum triterpenes screened by the response surface methodology are accurate and reliable, and can be used for the processing and preparation of Ganoderma lucidum triterpenes. The active triterpenoid components in Ganoderma lucidum can inhibit lipoxygenase, indicating their abilities to relieve inflammation. The research results can provide a theoretical basis for further studies on the anti-inflammatory components in Ganoderma lucidum.
