基于特征图谱和多成分含量测定的茯苓质量评价研究
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摘要
该文建立了茯苓三萜类成分的HPLC特征图谱,并同时测定16α-羟基松苓新酸、茯苓新酸B、去氢土莫酸、茯苓新酸A、猪苓酸C、茯苓新酸AM、3-O-乙酰-16α-羟基松苓新酸、去氢茯苓酸、茯苓酸、松苓新酸10个三萜类成分的含量。采用Welch Ultimate XB C_(18)色谱柱(4.6 mm×250 mm,5μm);流动相为乙腈(含3%四氢呋喃)(A)和0.1%甲酸水溶液(B),梯度洗脱,流速1.0 mL·min~(-1);柱温30℃;检测波长201,243 nm;进样量20μL。采用SPSS 22.0及GraphPad Prism 7.0对茯苓药材与饮片、不同产地茯苓样品数据进行分析。结果显示,所建立的特征图谱专属性强,10个待测成分能够达到良好分离,在考察的范围内线性关系良好(r≥0.999 6),平均回收率98.53%～103.8%,RSD 1.7%～2.7%。该方法重复性好,专属性强,可用于茯苓的鉴别及质量控制。研究结果表明,茯苓中10个三萜类成分之间具有正相关;产地收集的药材中各成分的含量均高于市售饮片;湖北、云南产茯苓10个成分的总含量略高于安徽,但安徽产茯苓中各成分的含量波动范围较小。
HPLC specific chromatograms of Poria were established, and the concentrations of 10 triterpenoids(16α-hydroxydehydrotrametenolic acid, poricoic acid B, dehydrotumulosic acid, poricoic acid A, polyporenic acid C, poricoic acid AM, 3-O-acetyl-16α-hydroxydehydrotrametenolic acid, dehydropachymic acid, pachymic acid, and dehydrotrametenolic acid) were simultaneously determined. Chromatographic analysis was conducted on a Welch Ultimate XB C_(18) column(4.6 mm × 250 mm,5 μm). Acetonitrile solution(contain 3% tetrahydrofuran)(A) and 0.1% formic acid aqueous solution(B) were used as the mobile phase with gradient elution at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃ and the injection volume was 20 μL. The experimental data were analyzed by the SPSS 22.0 and GraphPad Prism 7.0. The established triterpenoids fingerprints were specific, and the 10 components were well separated and showed good linearity(r≥0.999 6) within the concentration ranges tested. The mean recoveries were between 98.53%-103.8%(RSD 1.7%-2.7%). The method was specific and repeatable, and could be used for identification and quality evaluation of Poria. The results showed that the contents of 10 triterpenoids were positively correlated with each other. The contents of 10 triterpenoids of samples collected from producing areas were higher than that collected from markets. The total contents of 10 triterpenoids of samples collected from Hubei and Yunnan province were slightly higher than that from Anhui province, but the contents of samples from Anhui province were varied in smaller ranges.
HPLC specific chromatograms of Poria were established, and the concentrations of 10 triterpenoids(16α-hydroxydehydrotrametenolic acid, poricoic acid B, dehydrotumulosic acid, poricoic acid A, polyporenic acid C, poricoic acid AM, 3-O-acetyl-16α-hydroxydehydrotrametenolic acid, dehydropachymic acid, pachymic acid, and dehydrotrametenolic acid) were simultaneously determined. Chromatographic analysis was conducted on a Welch Ultimate XB C_(18) column(4.6 mm × 250 mm,5 μm). Acetonitrile solution(contain 3% tetrahydrofuran)(A) and 0.1% formic acid aqueous solution(B) were used as the mobile phase with gradient elution at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃ and the injection volume was 20 μL. The experimental data were analyzed by the SPSS 22.0 and GraphPad Prism 7.0. The established triterpenoids fingerprints were specific, and the 10 components were well separated and showed good linearity(r≥0.999 6) within the concentration ranges tested. The mean recoveries were between 98.53%-103.8%(RSD 1.7%-2.7%). The method was specific and repeatable, and could be used for identification and quality evaluation of Poria. The results showed that the contents of 10 triterpenoids were positively correlated with each other. The contents of 10 triterpenoids of samples collected from producing areas were higher than that collected from markets. The total contents of 10 triterpenoids of samples collected from Hubei and Yunnan province were slightly higher than that from Anhui province, but the contents of samples from Anhui province were varied in smaller ranges.
引文
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[12] 车爽,李清,霍艳双,等. 波长转换RP-HPLC法同时测定茯苓不同部位中5种三萜酸含量[J]. 药学学报,2010,45(4):494.
[13] 沈玉萍,杨欢,陈斌,等. 反相高效液相色谱法测定中药茯苓中4种三萜酸的含量[J]. 中国药学杂志,2011,46(5):388.
[14] 彭灿,余生兰,张静,等. HPLC同时测定茯苓中4种三萜酸的含量[J]. 中药材,2017,40(7):1643.
[15] 张靓琦,贾英,罗洁,等. 超高效液相色谱法同时测定茯苓中去氢土莫酸等6种活性成分的含量[J]. 中国药学杂志,2012,47(13):1080.
[16] 王志进. 谈提高大别山区茯苓饮片加工质量[J]. 中国中药杂志,1992(5):280.
[17] 池秀莲,杨光,马帅,等. 我国茯苓国际贸易研究与问题探析[J]. 中国中药杂志,2018,43(1):191.
[18] 程磊,侯俊玲,王文全,等. 我国茯苓生产技术现状调查分析[J]. 中国现代中药,2015,17(3):195.
[2] 王克勤,傅杰,苏玮,等. 道地药材茯苓疏[J]. 中药研究与信息,2002(6):16.
[3] 杨树东,包海鹰. 茯苓中三萜类和多糖类成分的研究进展[J]. 菌物研究,2005(3):55.
[4] 沈玉萍,李军,贾晓斌. 中药茯苓化学成分的研究进展[J]. 南京中医药大学学报,2012,28(3):297.
[5] 黎红,许津. 茯苓素的反相HPLC分析法[J]. 中国抗生素杂志,1993(6):474.
[6] 冯亚龙,赵英永,丁凡,等. 茯苓皮的化学成分及药理研究进展(Ⅰ)[J]. 中国中药杂志,2013,38(7):1098.
[7] 仲兆金,刘浚. 茯苓有效成分三萜的研究进展[J]. 中成药,2001(1):60.
[8] 韩小娟,罗建光,陆园园,等. HPLC-DAD-ESI-MSn法鉴定茯苓中的三萜成分[J]. 药学与临床研究,2009,17(4):290.
[9] 康安,郭锦瑞,谢彤,等. UPLC-LTQ-Orbitrap质谱联用技术分析茯苓中的化学成分[J]. 南京中医药大学学报,2014,30(6):561.
[10] 李珂,张立群,聂晶. 不同药用部位茯苓UPLC-UV-MS指纹图谱比较研究[J]. 中药材,2013,36(3):382.
[11] 昝俊峰,徐斌,刘军锋,等. 茯苓三萜成分RP-HPLC-ELSD指纹图谱研究[J]. 中草药,2012,43(5):895.
[12] 车爽,李清,霍艳双,等. 波长转换RP-HPLC法同时测定茯苓不同部位中5种三萜酸含量[J]. 药学学报,2010,45(4):494.
[13] 沈玉萍,杨欢,陈斌,等. 反相高效液相色谱法测定中药茯苓中4种三萜酸的含量[J]. 中国药学杂志,2011,46(5):388.
[14] 彭灿,余生兰,张静,等. HPLC同时测定茯苓中4种三萜酸的含量[J]. 中药材,2017,40(7):1643.
[15] 张靓琦,贾英,罗洁,等. 超高效液相色谱法同时测定茯苓中去氢土莫酸等6种活性成分的含量[J]. 中国药学杂志,2012,47(13):1080.
[16] 王志进. 谈提高大别山区茯苓饮片加工质量[J]. 中国中药杂志,1992(5):280.
[17] 池秀莲,杨光,马帅,等. 我国茯苓国际贸易研究与问题探析[J]. 中国中药杂志,2018,43(1):191.
[18] 程磊,侯俊玲,王文全,等. 我国茯苓生产技术现状调查分析[J]. 中国现代中药,2015,17(3):195.