高效液相色谱法同时测定红景天药材中6种有效成分的含量及质量相关性分析

Simultaneous Determination of 6 Effective Ingredients in Rhodiola by High Performance Liquid Chromatography and Quality Correlation Analysis

将1.0 g红景天药材粉末用10 mL甲醇超声提取30 min,过滤后即得供试品溶液.以ZORBAX SB-C_(18)色谱柱为固定相,以不同体积比的甲醇和0.3%(体积分数)磷酸溶液的混合液为流动相进行梯度洗脱,采用高效液相色谱法同时测定供试品溶液中6种有效成分的含量,0~50 min时,在275 nm检测波长下检测红景天苷、熊果苷、酪醇、没食子酸,50~75 min时,在360 nm检测波长下检测山萘酚、槲皮素.结果表明,6种有效成分的质量在一定范围内与其对应的峰面积呈线性关系,检出限(3S/N)为0.02~0.04 mg·g^(-1).按标准加入法进行回收试验,回收率为96.8%~99.7%.在精密度试验中,测定值的相对标准偏差(n=6)均小于2.0%.方法用于分析5种红景天药材各3批,其中红景天苷的含量均符合药典规定.方差分析和q检验结果表明,5种红景天药材中红景天苷、熊果苷、酪醇、山萘酚、槲皮素含量有显著性差异,没食子酸含量无显著性差异,其中大花红景天中红景天苷、熊果苷、山萘酚、槲皮素的含量最高,高山红景天的酪醇含量最高,长鞭红景天中红景天苷含量最低.

The Rhodiola powder(1.0 g)was extracted ultrasonically for 30 min in 10 mL of methanol,and the test solution was obtained after filtration.Six effective ingredients in the test solution were determined by high performance liquid chromatography with the detection wavelengths of 275 nm for salidroside,arbutin,tyrosol and gallic acid in 0-50 min,and 360 nm for kaempferol and quercetin in 50-75 min,using ZORBAX SB-C_(18) column as the stationary phase,and a mixture of methanol and 0.3%(φ)phosphate solution in different volume ratios as the mobile phase for gradient elution.As shown by the results,linear relationships between values of peak area and mass of six effective ingredients were found in definite ranges,with detection limits(3 S/N)ranged from 0.02 mg·g^(-1) to 0.04 mg·g^(-1).Test for recovery was made by standard addition method,giving results in the range of 96.8%-99.7%.RSDs(n=6)of the determined values were less than 2.0%in precision test.This method was applied to analysis of 5 species of Rhodiola(3 batches in one species),and the contents of salidroside were in line with the specification of pharmacopoeia.The results of analysis of variance and q-test showed that there were significant differences in the contents of salidroside,arbutin,tyrosol,kaempferol and quercetin,but there was no significant difference in the content of gallic acid.Among them,the contents of salidroside,arbutin,kaempferol and quercetin in Rhodiola crenulata were the highest,the content of tyrosol in Rhodiola cretinii was the highest,and the content of salidroside in Rhodiola fastigiata was the lowest.