珠子草药材质量标准及HPLC指纹图谱研究
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摘要
目的建立珠子草药材的质量标准及HPLC指纹图谱,探讨不同产地珠子草药材成分的差异性。方法分别从性状、鉴别、检查和浸出物、含量测定方面对珠子草药材的质量标准进行研究。采用ZORBAX Eclipse plus C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%磷酸水溶液作为流动相进行梯度洗脱,柱温25℃,检测波长270 nm,流速1.0 mL/min,进样量10μL,对10个不同产地珠子草药材进行测定;采用相似度分析和主成分分析对指纹图谱进行评价。结果拟定了珠子草药材质量标准草案,建立了10个不同产地珠子草药材的HPLC指纹图谱,确定了8个共有峰,指认了短叶苏木酚和柯里拉京2个特征峰。不同产地珠子草的化学成分组成及含量存在一定差异,其中柯里拉京可作为区别产地的特征性物质。结论本研究建立的定性定量检测方法操作简便、准确可靠、专属性强、重复性好,可作为珠子草药材的质量控制方法;建立的珠子草HPLC指纹图谱可为该药材的鉴别及质量控制提供参考。
Objective To establish the quality standards and HPLC fingerprints of Phyllanthus niruri Linn.; To study the differences in the components of Phyllanthus niruri Linn. from different producing areas. Methods The quality standards of Phyllanthus niruri Linn. was studied from characters, identification, inspection with extract, and content determination. The HPLC separation was performed on ZORBAX Eclipse plus C18 column(250 mm × 4.6 mm,5 μ m). The mobile phase was acetonitrile-0.1% phosphoric acid water in gradient elution; the column temperature was25 ℃; the detection wavelength was 270 nm; the flow rate was 1.0 m L/min; the sample volume was 10 μ L.Phyllanthus niruri Linn. from ten different producing areas were determined; similarity analysis and principal component analysis were used to analyze the fingerprints. Results The quality standard of Phyllanthus niruri Linn.was drafted. The HPLC fingerprints of Phyllanthus niruri Linn. from ten different producing areas were established with eight common peaks, and two characteristic peaks included brevifolincarboxylic acid and corilagin were confirmed. There were some differences in the chemical components and content of Phyllanthus niruri Linn. from different producing areas, of which corilagin could be used as the characteristic material to distinguish the difference of producing areas. Conclusion The qualitative and quantitative determination method is simple, accurate and reliable, with good stability and reproducibility, which can be used for the quality control of Phyllanthus niruri Linn.The established HPLC fingerprints can be used for the identification and quality control of Phyllanthus niruri Linn.
Objective To establish the quality standards and HPLC fingerprints of Phyllanthus niruri Linn.; To study the differences in the components of Phyllanthus niruri Linn. from different producing areas. Methods The quality standards of Phyllanthus niruri Linn. was studied from characters, identification, inspection with extract, and content determination. The HPLC separation was performed on ZORBAX Eclipse plus C18 column(250 mm × 4.6 mm,5 μ m). The mobile phase was acetonitrile-0.1% phosphoric acid water in gradient elution; the column temperature was25 ℃; the detection wavelength was 270 nm; the flow rate was 1.0 m L/min; the sample volume was 10 μ L.Phyllanthus niruri Linn. from ten different producing areas were determined; similarity analysis and principal component analysis were used to analyze the fingerprints. Results The quality standard of Phyllanthus niruri Linn.was drafted. The HPLC fingerprints of Phyllanthus niruri Linn. from ten different producing areas were established with eight common peaks, and two characteristic peaks included brevifolincarboxylic acid and corilagin were confirmed. There were some differences in the chemical components and content of Phyllanthus niruri Linn. from different producing areas, of which corilagin could be used as the characteristic material to distinguish the difference of producing areas. Conclusion The qualitative and quantitative determination method is simple, accurate and reliable, with good stability and reproducibility, which can be used for the quality control of Phyllanthus niruri Linn.The established HPLC fingerprints can be used for the identification and quality control of Phyllanthus niruri Linn.
引文
[1]程新平,牛晓峰,朱周才.珠子草生药学研究[J].西安医科大学学报(医学版),2001,22(2):162-163.
[2]朱宇同,李秉滔,方宏勋.珠子草新亚种的发现和鉴定[J].广州中医药大学学报,1997,14(2):119-121.
[3]KODAKANDLA V S,BIKRAM S,RAGHUNATH S T,et al.Antihepatotoxic princinple of Phyllanthus niruri herbs[J].Journal of Ethnopharmacoligy,1985,14(l):41-44.
[4]UENO H,HORIE S,NISHI Y,et al.Chemical and pharmaceutical studies on medicinal plants in Paraguay.Garaniin,an angiotensin-converting anzyme inhibitor from“paraparai mi,”Phyllanthus niruri[J].Journal of Natural Products,1988,51(2):357-359.
[5]SOMANABANDHU A.1H-and 13C-nmr assignment of phyllanthin and hypophyllanthin:lingnans that enhance cytotoxic responses with cultured multidrug-resistant cells[J].Journal of Natural Products,1993,56(2):261-267.
[6]SNATOS A R.Analgesic effects of callus culture extracts from selected species of Phyllanthus in mice[J].Journal of Pharmacy and Pharmacology,1994,46(9):755-759.
[7]CIMANGA R K,TONA L,LUYINDULA N,et al.In vitro antiplasmodial activity of callus culture extracts and fractions from fresh apical stems of Phyllanthus niruri L.(Euphorbiaceae):part 2[J].Journal of Ethnopharmacoligy,2004,95(2/3):399-404.
[8]李彩东,吴斌,杨静.珠子草的生药鉴定[J].中药材,2000,23(9):522-523.
[9]KARD D.Evaluation of trace metal concentration in some herbs and herbal teas by principal component analysis[J].Journal of Food Chemistry,2009,114(1):347-354.
[2]朱宇同,李秉滔,方宏勋.珠子草新亚种的发现和鉴定[J].广州中医药大学学报,1997,14(2):119-121.
[3]KODAKANDLA V S,BIKRAM S,RAGHUNATH S T,et al.Antihepatotoxic princinple of Phyllanthus niruri herbs[J].Journal of Ethnopharmacoligy,1985,14(l):41-44.
[4]UENO H,HORIE S,NISHI Y,et al.Chemical and pharmaceutical studies on medicinal plants in Paraguay.Garaniin,an angiotensin-converting anzyme inhibitor from“paraparai mi,”Phyllanthus niruri[J].Journal of Natural Products,1988,51(2):357-359.
[5]SOMANABANDHU A.1H-and 13C-nmr assignment of phyllanthin and hypophyllanthin:lingnans that enhance cytotoxic responses with cultured multidrug-resistant cells[J].Journal of Natural Products,1993,56(2):261-267.
[6]SNATOS A R.Analgesic effects of callus culture extracts from selected species of Phyllanthus in mice[J].Journal of Pharmacy and Pharmacology,1994,46(9):755-759.
[7]CIMANGA R K,TONA L,LUYINDULA N,et al.In vitro antiplasmodial activity of callus culture extracts and fractions from fresh apical stems of Phyllanthus niruri L.(Euphorbiaceae):part 2[J].Journal of Ethnopharmacoligy,2004,95(2/3):399-404.
[8]李彩东,吴斌,杨静.珠子草的生药鉴定[J].中药材,2000,23(9):522-523.
[9]KARD D.Evaluation of trace metal concentration in some herbs and herbal teas by principal component analysis[J].Journal of Food Chemistry,2009,114(1):347-354.