HPLC法同时测定大叶冬青叶中6种有效成分的含量
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摘要
目的建立HPLC同时测定大叶冬青叶中地榆皂苷Ⅱ、坡模酸、27-p-Coumaroyloxy ursolic acid、胡萝卜苷、3β-Hydroxyurs-11-en-28,13β-olide、乌发醇6种三萜类成分含量的方法,并探讨不同产地及不同采收期大叶冬青叶中三萜类成分含量的差异。方法采用Agilent HC-C_(18)(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.4%磷酸水溶液为流动相,梯度洗脱,检测波长205 nm,柱温30℃,流速1.0 mL·min~(-1),对大青叶6种三萜类化合物进行含量测定。采用中药色谱指纹图谱相似度评价系统(2004A)软件进行指纹图谱分析。结果 6种三萜类化合物均有良好的分离度,呈现较好的线性关系,平均加样回收率为96.88%～105.43%,RSD均小于3%(n=6)。不同产地、不同采收期样品6种三萜类成分的含量差异较为明显。14批同产地不同采收期样品指纹图谱相似度大于0.9,表明样品一致性良好。10批不同产地样品指纹图谱相似度较低,存在明显差异。结论该方法准确、快速、具有良好的重复性,可用于同时测定大叶冬青叶中6种成分的含量;不同产地、不同采收期药材内在质量存在一定差异。
Objective An HPLC method was established for simultaneous determination of six triterpenoids in the leaves of Ilex latifolia. The contents of triterpenoids were compared in the samples from different origins with different harvesting time. Methods The analysis was performed on an Agilent HC-C_(18) column(4.6 mm × 250 mm,5 μm) with gradient elution of acetonitrile-0.4% phosphoric acid solution system at a flow rate of 1.0 mL·min~(-1) and a column temperature of 30 ℃. The detection wavelength was 205 nm. Fingerprint analysis was performed using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2004 A) software. Results The separation of six triterpenoids and linear relationships were good;the average recoveries were 96.88%-105.43%,and the RSDs were less than 3%(n=6). The contents of six triterpenoids in samples from different origins and different harvesting time were significantly different. The similarity of fingerprints of 14 batches of samples from same origin with different harvesting periods was more than 0.9,which indicated that the samples had good consistency.The similarity of fingerprints of 10 batches samples from different habitats was very poor with significant difference.Conclusion The method is accurate,rapid,repeatable and stable,which can be used for the determination of six triterpenoids in Ilex latifolia. There are some differences in the intrinsic quality of medicinal materials from different origins and different harvesting time.
Objective An HPLC method was established for simultaneous determination of six triterpenoids in the leaves of Ilex latifolia. The contents of triterpenoids were compared in the samples from different origins with different harvesting time. Methods The analysis was performed on an Agilent HC-C_(18) column(4.6 mm × 250 mm,5 μm) with gradient elution of acetonitrile-0.4% phosphoric acid solution system at a flow rate of 1.0 mL·min~(-1) and a column temperature of 30 ℃. The detection wavelength was 205 nm. Fingerprint analysis was performed using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2004 A) software. Results The separation of six triterpenoids and linear relationships were good;the average recoveries were 96.88%-105.43%,and the RSDs were less than 3%(n=6). The contents of six triterpenoids in samples from different origins and different harvesting time were significantly different. The similarity of fingerprints of 14 batches of samples from same origin with different harvesting periods was more than 0.9,which indicated that the samples had good consistency.The similarity of fingerprints of 10 batches samples from different habitats was very poor with significant difference.Conclusion The method is accurate,rapid,repeatable and stable,which can be used for the determination of six triterpenoids in Ilex latifolia. There are some differences in the intrinsic quality of medicinal materials from different origins and different harvesting time.
引文
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[11]李宏杨,刘飞,张凤琴,等. 6种冬青科苦丁茶总三萜类物质含量的比较研究[J].热带生物学报,2010,1(1):17-20.
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[13]王存琴,王宏婷,许浩,等.大叶冬青叶的三萜类成分研究[J].中药材,2015,38(8):1654-1656.
[14]王存琴,王磊,李宝晶,等.大叶冬青的黄酮类成分研究[J].中国中药杂志, 2014,39(2):258-261.
[15]王存琴,王宏婷,陈娟,等.苦丁茶三萜类成分及药理作用研究进展[J].中国临床药理学与治疗学,2016,21(6):703-709.
[16]王存琴,张艳华.大叶冬青的化学成分及药理活性研究进展[J].中国药房,2014,25(11):1052-1054.
[2]易帆,彭勇,许利嘉,等.大叶苦丁茶的研究进展[J].中国现代中药,2013,15(8):710-717.
[3]谭银丰,李海龙,王群,等.大叶冬青降血脂作用及多糖含量测定研究[J].海南医学院学报,2011,17(1):24-26.
[4] LIU L, YI S, LAURA T, et al. Determination of polyphenolic content and antioxidant activity of kudingcha made from Ilex kudingcha,C. J. Tseng[J]. Journal of Tea Science,2008,112(1):35-41.
[5]陈凤美,李晓储,杨守辉,等.大叶冬青叶抗肿瘤活性初步研究[J].林业工程学报,2007,21(5):30-31.
[6]范春林.大叶冬青的化学成分研究[D].广州:暨南大学,2010.
[7] SHYU M H,KAO T C,YEN G C. Oleanolic acid and ursolic acid induce apoptosis in HuH7 human hepatocellular carcinoma cells through a mitochondrial-dependent pathway and downregulation of XIAP[J]. Journal of Agricultural&Food Chemistry,2010,58(10):6110-6118.
[8]王宏婷,王存琴. 27-O-(E)-香豆酰基-乌索酸通过调控JNK/SAPK通路诱导人乳腺癌细胞MDA-MB-231细胞凋亡[J].中国中药杂志,2015,40(4):722-726.
[9] WANG C Q,WANG L,FAN C L,et al. Ilelic acids A and B,two unusual triterpenes with a seven-membered ring from Ilex latifolia[J]. Organic Letters,2012,14(16):4102-4105.
[10]张俊清,谭银丰,李海龙,等.海南大叶冬青中2种活性成分的含量研究[J].中国药学杂志,2010,45(9):658-660.
[11]李宏杨,刘飞,张凤琴,等. 6种冬青科苦丁茶总三萜类物质含量的比较研究[J].热带生物学报,2010,1(1):17-20.
[12] WANG C Q,LI M M,ZHANG W,et al. Four new triterpenes and triterpene glycosides from the leaves of Ilex latifolia,and their inhibitory activity on triglyceride accumulation[J]. Fitoterapia,2015,106:141-146.
[13]王存琴,王宏婷,许浩,等.大叶冬青叶的三萜类成分研究[J].中药材,2015,38(8):1654-1656.
[14]王存琴,王磊,李宝晶,等.大叶冬青的黄酮类成分研究[J].中国中药杂志, 2014,39(2):258-261.
[15]王存琴,王宏婷,陈娟,等.苦丁茶三萜类成分及药理作用研究进展[J].中国临床药理学与治疗学,2016,21(6):703-709.
[16]王存琴,张艳华.大叶冬青的化学成分及药理活性研究进展[J].中国药房,2014,25(11):1052-1054.