HPLC法测定枳壳中6个二氢黄酮类成分的含量

目的:建立HPLC法测定枳壳药材中6个黄酮类成分圣草次苷、新圣草苷、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷的含量。方法:采用Inertsil ODS-4(4.6 mm×150 mm, 5 μm)色谱柱, 流动相为乙腈-0.1%磷酸(18∶82), 流速1.0 mL·min^-1, 检测波长283 nm, 柱温30℃。结果:圣草次苷、新圣草苷、芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷6个二氢黄酮类化合物的质量分别在0.02～0.16、0.60～0.54、0.02～0.18、0.41～3.70、0.16～1.40、0.22～1.99 μg范围内与峰面积呈良好的线性关系, r分别为0.9995、0.9999、0.9999、0.9999、0.9999和0.9999;平均加样回收率(n=6)分别为100.8%、102.5%、97.8%、99.6%、98.0%、102.7%。结论:含量测定方法可作为中药枳壳的质量控制的有效方法。

Objective:To establish an HPLC method for simultaneous determination of eriocitrin, neoeriocitrin, narirutin,naringin,hesperidin, neohesperidin content in Aurantii Fructus.Methods:HPLC separation was carried out with an Inertsil ODS-4 column(4.6 mm×150 mm,5 μm)using acetonitrile-0.1% phosphoric acid(18:82)as mobile phase at a flow rate of 1.0 mL·min^-1. The detection wavelength was 283 nm and the column temperature was maintained at 30℃.Results: Good linear relationships were observed between the masses and the peak areas of eriocitrin,neoeriocitrin,narirutin,naringin,hesperidin,neohesperidin in the ranges of 0.02-0.16(r=0.9995), 0.60-0.54 (r=0.9999),0.02-0.18(r=0.9999),0.41-3.70(r=0.9999),0.16-1.40(r=0.9999) and 0.22-1.99(r=0.9999)μg,respectively; The average recovery rates(n=6)were 100.8%, 102.5%, 97.8%,99.6%,98.0% and 102.7%,respectively.Conclusion:The established quantification method is applicable for the quality control of Aurantii Fructus.

1 ChP(中国药典).2010.VolⅠ(一部):229
2 CAO Jun(曹君),WANG Shao-jun(王少军),GONG Qian-feng(龚千锋),et al. Analysis of volatile oil in pre- and post-processed pieces of Fructus Aurantii from different habitats by GC-MS(不同产地枳壳饮片炮制前后挥发油成分的GC-MS分析).Chin Tradit Herb Drugs(中草药),2005,36(2):172
3 Kawaii S,Tomono Y,Katase E,et al.Quantitation of flavonoid constituents in Citrus Fruits.J Agric Food Chem,1999,47(9):3565
4 Sakata K,Hirose Y,Qiao Z,et al.Inhibition of inducible isoforms of cyclooxygenase and nitric oxide synthase by favonoid hesperidin in mouse macrophage cell line.Cancer Lett,2003,199(2):139
5 Garg A,Garg S,Zaneveld LJD,et al.Chemistry and pharmacology of the Citrus bioflavonoid hesperidin.Phytother Res,2001,15(8):655
6 Lee JH.Antiviral activity of some flavonoids on Heroes simplex virus.Korean Pharmacog,1999,30(1):49
7 Miyake Y,Yamamoto K,Morimitsu Y,et al.Isolation of C-glucosylfavone from lemon peel and antioxidative activity of favonoid compounds in lemon fruit.J Agric Food Chem,1997,45(12): 4619
8 Majo DD,Giammanco M,Guardia ML,et al.Flavanones in Citrus fruit: Structure-antioxidant activity relationships.Food Res Int,2005,38(10):1161
9 Yu J,Wang L,Walzem RL,et al.Antioxidant activity of Citrus limonoids,flavonoids,and coumarins.J Agric Food Chem,2005,53(6),2009
10 Finotti E,Di Majo D.InfUence of solvents on the antioxidant property of favonoids.Die Nahrung,2003,47(3):186
11 ZHANG Hong-wu(张宏武),FENG Yu-fei(封宇飞),ZOU Zhong-mei(邹忠梅).Simultaneous determination of four flavonoids in Citrus aurantium L.by HPLC(高效液相色谱法同时测定枳壳饮片中4种黄酮类化合物的含量).Chin J New Drugs(中国新药杂志),2008,17(18):1600
12 Ding L,Luo X,Tang F,et al.Simultaneous determination of favonoid and alkaloid compounds in Citrus herbs by high-performance liquid chromatography–photodiode array detection-electrospray mass spectrometry.J Chromatogr B,2007,857(2):202
13 ZHANG Jin-lian(张金莲),HE Min(何敏),et al.Determination of content of narigin and hesperidin and neohesperidin in Fructus Aurantii pieces by HPLC(高效液相色谱法测定枳壳饮片中柚皮苷、橙皮苷和新橙皮苷的含量).Chin J Exp Tradit Med Form(中国实验方剂学杂志),2010,16(6):68
14 Wistuba D,Bogdanski A,Larsen KL,et al.δ-Cyclodextrin as novel chiral probe for enantiomeric separation by electromigration methods.Electrophoresis,2006,27(21):4359
15 CHEN Hai-fang(陈海芳),WEI Ling(魏玲),YUAN Jin-bin(袁金斌),et al.RP-HPLC determination of five lipophilic components in Fructus Aurantii(RP-HPLC法测定枳壳中5个脂溶性成分的含量).Chin J Pharm Anal(药物分析杂志),2010,30(9):1610
16 XU Huan(徐欢),CHEN Hai-fang(陈海芳),JIE Lei(介磊),et al. HPLC determination of nobiletin and tangeretin in Fructus Aurantii(HPLC法测定枳壳中川陈皮素、红橘素的含量).Chin J Pharm Anal(药物分析杂志),2009,29(9):1411
17 HU Liu(胡柳),LUAN Lian-jun(栾连军),SHAO Qing(邵青),et al.Quantitative analysis and fingerprint study for quality evaluating of Chinese herb Fructus Aurantii[含量测定与特定(指纹)图谱分析相结合方法用于枳壳药材质量分析].Chin J Pharm Anal(药物分析杂志),2004,24(4):345