To optimize ganoderic acid analysis of the ultraviolet absorption method and colorimetric method, ultraviolet-visible spectrum was used to examine difference among ganoderic acids (A, C, C2 and E), lucidenic acid A and thymol, and among ursolic acid, oleanolic acid, and ganoderic acid A after the reaction with the color-developing agents vanillin and perchloric acid. After the extraction, the ganoderic acid contents were analyzed and calculated using the two methods taking the mycelium and fruiting bodies of Ganoderma lucidum as test samples based on different standard curves. The results showed that ganoderic acids A, C, C2, E, and lucidenic acid A had similar spectrum with the formation of a peak with the maximum absorbance at around 254 nm. However, thymol formed two peaks, and the trough was clearly observed at 246 nm. As to the vanillin-perchloric acid colorimetric assay, ursolic acid had similar spectrum with oleanolic acid with peaks at 544 nm and 552 nm, whereas almost no absorbance was recorded for ganoderic acid A, indicating that it could not be used as a standard material in the colorimetric assay. With ganoderic acid A and thymol as standards, the ultraviolet absorption method indicated that content of ganoderic acid was 0.7 mg/100 mg (dry biomass) and 1.3 mg/100 mg respectively in the mycelium, and 2.1 mg/100 mg and 4.0 mg/100 mg in the fruit body. Based on ursolic acid and oleanolic acid standard curves, the colorimetric assay showed that ganoderic acid content was 0.45 mg/100 mg and 0.49 mg/100 mg respectively in the mycelium, and 1.55 mg/100 mg and 1.71 mg/100 mg in the fruit body. By comparing these results, it can be concluded that there were significant differences in ganoderic acid content between the ultraviolet absorbance and the colorimetric methods and among different standards. The present study is valuable for optimization of ganoderic acid quantitative analysis.
Key words
Ganoderma lucidum; ganoderic acid; ultraviolet-visible spectrum; ultraviolet absorbance method; vanillin-perchloric acid colorimetric method
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