UPLC法测定羊红膻药材中2种黄酮苷类成分的含量

目的：采用超高效液相色谱法测定羊红膻中木犀草素-7-O-β-D-葡萄糖醛酸苷和芹菜素-7-O-β-D-葡萄糖醛酸苷2种黄酮苷的含量。方法：采用BEH C₁₈色谱柱（2.1 mm×100 mm，1.7μm），以乙腈-0.1%甲酸水溶液为流动相，梯度洗脱，流速0.4 mL·min^-1，柱温30℃，检测波长345 nm。结果：木犀草素-7-O-β-D-葡萄糖醛酸苷质量浓度在19.38~487.6 μg·mL^-1范围内具有良好线性关系（r=0.999 6，n=5），3批羊红膻药材中其含量分别为4.62、4.23、3.88 mg·g^-1；芹菜素-7-O-β-D-葡萄糖醛酸苷质量浓度在21.16~423.2 μg·mL^-1的范围内具有良好线性关系（r=0.999 7，n=5），3批羊红膻药材中其含量分别为3.81、3.36、4.01 mg·g^-1。结论：此方法可用于药材羊红膻中上述2个黄酮苷成分的含量测定，测定结果可为该药材质量标准的完善提供依据。

Objective: To establish an UPLC method for determination of two flavonoid glycosides luteolin-7-O-β-D-glucuronopyranoside and apigenin-7-O-β-D-glucuronopyranoside in Yanghongshan the whole plant or the radix of Pimpinella thellungiana Wolff.Methods: The chromatographic separation was performed at 30℃ on a BEH C₁₈ column(2.1 mm×50 mm, 1.7 μm). The mobile phase consisted of acetonitrile and 0. 1% formic acid water using a gradient elution. The flow rate was set at 0. 4 mL·min^-1 with the detection wavelength of 345 nm.Results: The standard curve of the luteolin-7-O-β-D-glucuronopyranoside showed good linearity in the range of 19.38 to 487.6 μg·mL^-1(r=0. 999 6). The content of 3 batches of Yanghongshan was 4. 62 mg·g^-1,4. 23 mg·g^-1 and 3.88 mg·g^-1 for luteoli-7-O-β-D-glucuronopyranoside,respectively. The standard curve of the apigenin-7-O-β-D-glucuronopyranoside was linear in the range of 21. 16 to 423. 2 μg·mL^-1(r=0. 999 7)and the contents of 3 batches of Yanghongshan were 3. 81 mg·g^-1,3. 36 mg·g^-1 and 4. 01 mg·g^-1 for apigenin-7-O-β-D-glucuronopyranoside, respectively.Conclusion: The established method can be used for the determination of flavonoid glycosides in Yanghongshan. Moreover,the quantitative results may be useful for the quality improvement of this herbal medicine.

[1] 金振国,王忙生.草药羊红膻的研究进展[J].商洛学院学报, 2008, 22(5):43 JIN ZG, WANG MS. Advances in studies of herbal of Pimpinella thellungiana Wolff[J]. J Shangluo Univ,2008, 22(5):43
[2] 陈忠孝,邵富生,石会丽.浅谈草药羊红膻[J].陕西中医, 1999, 20(12):566 CHEN ZX, SHAO FS, SHI HL. Preliminary discussion on Yanghongshan[J]. Shaanxi J Tradit Chin Med,1999,20(12):566
[3] 中国药典2015年版.一部[S]. 2015:750 ChP 2015. Vol Ⅰ[S]. 2015:750
[4] 王长岱,丁凯,郭五保.羊红膻化学成分的研究[J].陕西新医药,1980, 9(8):49 WANG CD, DING K, GUO WB. Studies on the chemical constituents of Yanghongshan(Ⅰ)[J]. Shaanxi Med J, 1980, 9(8):49
[5] 许青媛,沈雅琴,王德华.羊红膻黄酮苷药理作用研究[J].中草药,1994, 25(1):51 XU QY, SHEN YQ,WANG DH. Studies on pharmacological action of flavonoid glycoside in Yanghongshan[J]. Chin Tradit Herb Drugs,1994, 25(1):51
[6] 石会丽,冯一凡,刘珍,等.羊红膻药材中总黄酮的含量测定方法研究[J].西北药学杂志, 2009, 24(1):18 SHI HL, FENG YF, LIU Z, et al. Content determination of total flavonoids in Yanghongshan[J]. Northwest Pharm J,2009,24(1):18
[7] 石会丽,杨智锋,李富贤.羊红膻的显微及薄层鉴别研究[J].西北药学杂志, 2013, 28(5):453 SHI HL,YANG ZF,LI FX. The microscopic identification and TLC of Yanghongshan[J]. Northwest Pharm J,2013, 28(5):453
[8] 马思萌,刘睿,任晓亮,等. HPLC-DAD法测定苦碟子中木犀草素7-O-β-D-葡萄糖苷、木犀草素7-O-β-D-葡萄糖醛酸苷和芹菜素7-O-β-D-葡萄糖醛酸苷[J].现代药物与临床,2014,29(4):377 MA SM,LIU R,REN XL,et al. Determination of luteolin-7-O-β-D-glucopyranoside, luteolin-7-O-β-D-glucuronopyranosid and apigenin-7-O-β-D-glucuronopyranoside in Ixeris sonchifolia by HPLC-DAD[J]. Drugs Clin,2014, 29(4):377