野生和栽培滇龙胆主要活性组分含量特征及质量评价

目的: 建立滇龙胆高效液相色谱(HPLC)图谱,测定有效成分含量,结合化学计量学对野生和栽培药材进行质量评价。方法: 采用Shim-pack VP-ODS液相色谱柱(150 mm×4.6 mm,5 μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0~5 min,5%B;5~10 min,5%B→10%B;10~26 min,10%B→26%B;26~30 min,26%B→35%B),流速1.00 mL·min^-1;柱温30 ℃;检测波长241 nm,进样量5 μL。采用系统聚类,相似度分析和偏最小二乘判别对化学数据进行统计分析。结果: 27批药材马钱苷酸含量为5.375 mg·g^-1(S12)~20.234 mg·g^-1(S3),獐牙菜苦苷含量为2.246 mg·g^-1(S7)~9.653 mg·g^-1(S18),龙胆苦苷含量为24.252 mg·g^-1(S19)~128.350 mg·g^-1(S25),当药苷含量为0.510 mg·g^-1(S12)~3.498 mg·g^-1(S8);当药苷在4种活性成分中含量最低;所有样品依照4种化学成分的含量特征可分为a、b、c 3类;a类包括大部分野生样品和栽 培2年的滇龙胆,贵州的样品(S19)被单独归为b类,c类主要为3年生样品;相似度分析显示大部分样品相似度大于0.930,野生和栽培样品化学成分种类相似,但含量有明显变化;以14个共有峰峰面积为变量,进行偏最小二乘判别分析,建立PLS-DA模型,结果显示野生和栽培样品能被正确区分;栽培3年滇龙胆环烯醚萜与裂环烯醚萜类成分总含量最高,质量稳定,品质最佳。结论: 高效液相色谱图谱与多指标化学成分定量相结合可为野生和栽培滇龙胆药材质量控制和评价提供科学依据。

Objective: To develop a method of HPLC chromatogram combined with chemometrics to evaluate quality of wild grown and cultivated Gentiana rigescens Franch.ex Hemsl. Methods: The Shim-pack VP-ODS C₁₈(150 mm×4.6 mm,particle size:5 μm)was adopted at a temperature of 30 ℃.The mobile phase was water with 0.1% formic acid(A)and acetonitrile(B).A linear solvent gradient was used(0-5 min,5%B;5-10 min,5%B→10%B;10-26 min,10%B→26%B;26-30 min,26%B→35%B).The flow rate was 1.00 mL·min^-1.The detection wavelength was 241 nm and the injection volume was 5 μL.Chemical data were analyzed by hierarchical cluster,similarity analysis and partial least squares discriminant analysis. Results: Loganic acid content of 27 batches samples ranged from 5.375 mg·g^-1(S12)to 20.234 mg·g^-1(S3). Swertiamarin content of samples ranged from 2.246 mg·g^-1(S7)to 9.653 mg·g^-1(S18).Gentiopicroside content ranged from 24.252 mg·g^-1(S19)to 128.350 mg·g^-1(S25). Sweroside content ranged from 0.510 mg·g^-1(S12)to 3.498 mg·g^-1(S8).The content of sweroside was the lowest in four bioactive compounds.All the samples could be classed into three groups(a,b,c) by content of four iridoids.Group a included most of the wild grown samples and cultivated samples grown for 2 years.Samples from Guizhou(S19)were classed into group b. Group c included cultivated samples grown for 3 years.Similarity analysis results showed similarity of most samples were over 0.930.Compound varieties of wild grown and cultivated G.rigescens Franch.ex Hemsl.were similar.However,contents of those compounds varied significantly.Partial least squares discriminant analysis combined common peaks' areas could distinguish wild grown samples from cultivated ones.G.rigescens Franch.ex Hemsl.grown for three years had the highest total content of bioactive compounds,with stable and the best medicinal materiol quality. Conclusion: HPLC chromatogram combined with determination of main bioactive compounds could provide reference for quality evaluation and control of wild grown and cultivated G.rigescens Franch.ex Hemsl.