山楂叶提取物的指纹图谱研究

Study of HPLC fingerprint of Hawthorn leaves extract

目的研究不同批次山楂叶提取物的HPLC指纹图谱，并测定绿原酸、牡荆素葡萄糖苷、牡荆素鼠李糖苷、芦丁、牡荆素、金丝桃苷和槲皮素的含量。方法采用AgilentZorbaxSB—C18色谱柱（250mm×4．6mm，5μm），0．5％甲酸-乙腈-甲醇-四氢呋喃溶液梯度洗脱，流速1．0mL·min^-1，柱温30℃，采用可变检测波长：0—15min时，检测波长260nm、参比波长360nm；15～65min时，检测波长370nm、参比波长430nm。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。结果绿原酸、牡荆素葡萄糖苷、牡荆素鼠李糖苷、芦丁、牡荆素、金丝桃苷和槲皮素的线性范围分别为11～360、19—600、32～1020、1．3—40、7．5～240、9．4～300、0．63—20μg·mL^-1，r均为0．9999；平均加样回收率、RSD分别为99．67％（0．88％）、100．17％（0．34％）、99．80％（0．18％）、100．00％（2．48％）、99．70％（0．45％）、100．17％（0．94％）和98．75％（1．16％）；18批山楂提取物的HPLC指纹图谱共标定出10个共有峰，并测定了其中7个成分的含量。结论该分析方法准确可靠，重复性好，能更好地控制山楂叶提取物的质量。

OBJECTIVE To study the HPLC fingerprint of Hawthorn leaves extract from different batches and to detect the contents of chlorogenic acid, glucosyl - vitexin, rhamnosyl - vitexin, rutin, vitexin, hyperoside and quercetin. METHODS With an Agilent Zorbax SB - C18 column（250 mm × 4.6 mm,5 μm）, gradientelution was performed with mobile phase of 0.5% formic acid - acetoni- trile -methanol -tetrahydrofuran and the wavelength was set at 0 - 15 min, and the detection wavelength was 260 nm, and the refer- ence wavelength was 360 nm, at 15 -65 min, the detection wavelength was 370 nm, the reference wavelength was 430 nm with the flow rate of 1.0 mL. min-land the column temperature was 30 ℃. The data calculation was performed with similarity evaluation system for chromatographic fingerprint of TCM （ Version 2004A）. RESULTS The linear range were 11 - 360, 19 - 600, 32 - 1020, 1.3 - 40, 7.5 - 240,9.4 - 300,0.63 - 20 μg· mL- l, respectively （ r = 0. 9999 ）. The average recovery rates were 99.67% （ RSD = 0.88% ） , 100. 17% （RSD = 0.34% ） ,99. 80% （RSD = 0.18% ）, 100.00% （RSD = 2.48% ） ,99.70% （RSD = 0.45% ）, 100.17% （RSD = 0.94% ） ,98.75% （ RSD = 1.16% ） , respectively. 10 common peaks were found from 18 batches of Hawthorn leaves extract HPLC fingerprint, of which seven peaks were confirmed. CONCLUSION This method is accurate, reproducible and provides a scientific basis for quality control of Hawthorn leaves extracts.