灵芝中10种灵芝酸类成分含量一测多评法的建立

Establishment of quantitative analysis of multi-components by single marker for content determination of 10 ganoderic acids in Ganoderma lucidum

目的建立同时测定灵芝中10种灵芝酸类成分的一测多评法。方法采用高效液相色谱(HPLC)法,以灵芝酸A为内参物,分别计算灵芝酸B、灵芝酸C_(2)、灵芝酸D、灵芝酸F、灵芝酸H、灵芝烯酸A、灵芝烯酸B、灵芝烯酸C、灵芝烯酸D 9种成分的相对校正因子,用相对校正因子计算上述灵芝酸类成分的含量,并与外标法测定结果进行比较。结果灵芝酸A、灵芝酸B、灵芝酸C_(2)、灵芝酸D、灵芝酸F、灵芝酸H、灵芝烯酸A、灵芝烯酸B、灵芝烯酸C、灵芝烯酸D的线性范围分别为0.032~3.996、0.040~4.971、0.037~4.568、0.028~3.558、0.033~4.177、0.044~5.440、0.032~3.944、0.040~4.994、0.045~5.593、0.035~4.342 mg/mL(R2均不小于0.9992);精密度、稳定性(24 h)、重复性试验的RSD均小于2%;平均加样回收率分别为99.43%、100.25%、98.50%、99.88%、100.59%、99.64%、98.50%、99.40%、99.64%、99.76%(RSD<2%,n=6)。灵芝酸B、灵芝酸C_(2)、灵芝酸D、灵芝酸F、灵芝酸H、灵芝烯酸A、灵芝烯酸B、灵芝烯酸C、灵芝烯酸D的相对校正因子平均值分别是1.7885、1.2882、1.1264、1.6985、0.8854、5.4681、4.2109、5.7808、4.2903;不同产地灵芝样品一测多评法所得含量与外标法的相对误差均在±12%之内。结论以灵芝酸A为内参物同时测定灵芝中10种灵芝酸类成分含量的一测多评法是可行的,该方法准确性和重复性良好,可用于灵芝的质量控制。

OBJECTIVE To establish a quantitative analysis of multi-components by single marker(QAMS)method for simultaneous determination of 10 ganoderic acids in Ganoderma lucidum.METHODS Using ganoderic acid A as internal reference,high-performance liquid chromatography(HPLC)method was adopted to calculate relative correction factors of the other 9 components,such as ganoderic acid B,ganoderic acid C_(2),ganoderic acid D,ganoderic acid F,ganoderic acid H,ganoderenic acid A,ganoderenic acid B,ganoderenic acid C,ganoderenic acid D;the contents of above ganoderic acids were calculated with relative correction factors,and compared with the results of external standard method.RESULTS The linear relationship of ganoderic acid A,ganoderic acid B,ganoderic acid C_(2),ganoderic acid D,ganoderic acid F,ganoderic acid H,ganoderenic acid A,ganoderenic acid B,ganoderenic acid C and ganoderenic acid D were 0.032-3.996,0.040-4.971,0.037-4.568,0.028-3.558,0.033-4.177,0.044-5.440,0.032-3.944,0.040-4.994,0.045-5.593 and 0.035-4.342 mg/mL(all R2≥0.9992),respectively.RSDs of precision,stability(24 h)and reproducibility tests were all lower than 2%.Their average recovery rates were 99.43%,100.25%,98.50%,99.88%,100.59%,99.64%,98.50%,99.40%,99.64%and 99.76%,respectively(RSD<2%,n=6).Relative correction factors of ganoderic acid B,ganoderic acid C_(2),ganoderic acid D,ganoderic acid F,ganoderic acid H,ganoderenic acid A,ganoderenic acid B,ganoderenic acid C and ganoderenic acid D were 1.7885,1.2882,1.1264,1.6985,0.8854,5.4681,4.2109,5.7808,4.2903,respectively.Relative errors between the content obtained by QAMS method and external standard method for G.lucidum from different origins were within±12%.CONCLUSIONS It is feasible that the contents of 10 ganoderic acids are determined simultaneously by QAMS method,using ganoderic acid A as internal reference.This method shows good precision and reproducibility and can be used for the quality control of G.lucidum.