HPLC双波长等基线融合结合校正因子法测定虎杖中3种成分的含量

Content Determination of 3 Components of Polygonum cuspidatum by HPLC Dual Wavelength Baseline Fusion Combined with Correction Factor

目的：为虎杖中白藜芦醇苷、大黄素、大黄素甲醚的含量测定提供更为简便、合理、可靠的方法。方法：采用高效液相色谱法。色谱柱为AgilentTC—C18（250mm×4．6mm，5gm），柱温为25℃，流动相为0．1％甲酸水溶液．乙腈（梯度洗脱），流速为1ml／min，检测波长为254nm（大黄素、大黄素甲醚）与319nm（白藜芦醇苷）。使用Matlab 7．1编程进行谱图融合；在此基础上，以虎杖中大黄素为内参物，建立其与白藜芦醇苷、大黄素甲醚的相对校正因子，计算白藜芦醇苷和大黄素甲醚的含量，并对不同产地虎杖中3种成分含量校正因子法的计算值与外标法实测值进行比较。结果：白藜芦醇苷、大黄素与大黄素甲醚的进样量分剐在0．29～1．76、0．75～4．51、0．05～0．31μg范围内与各自峰面积积分值呈良好的线性关系（r分别为0．9990、0．9998、0．9996）；精密度、重复性、稳定性试验的RSD〈3％；平均加样回收率分别为98．50％、101．68％、99．22％，RSD分别为1．81％、2．03％、2．44％（”均为6）。采用校正因子法与外标法测得不同产地虎杖中3种成分的含量差异无统计学意义。结论：该试验为虎杖的质量控制提供了一种更为简便、合理、可靠的方法。

OBJECTIVE： To establish a simple, reasonable and reliable method for the content determination of polydatin, emodin and physcion in Polygonum cuspidatum. METHODS：HPLC method was adopted The determination was performed on Agilent TC-C18（250 mm×4.6 mm, 5μm） column with mobile phase consisted of 0.1% formic acid-acetonitrile （gradient elution） at flow rate of 1 ml/min. The column temperature was 25℃ and the detection wavelengths were set at 254 nm （emodin, physcion） and 319 nm （polydatin）. Maflab 7.1 programming was used for chromatogram fusion; using emodin as internal standard, the relative correction factors of polydatin and physcion were established to calculate the contents of polydatin and physcion. The contents of 3 components from different producing areas calculated by correction factor method were compared with the measured values of external standard method. RESULTS： The linear ranges of polydatin, emodin and physcion were 0.29-1.76 μg（r=0.999 0）, 0.75-4.51μg （r=0.999 8） and 0.05-0.31μg（r=0.999 6）. RSDs of precision, reproducibility and stability tests were all lower than 3%. Average recovery rates were 98.50% （RSD= 1.81%, n=6）, 101.68% （RSD=2.03%, n=6） and 99.22% （RSD=2.44%, n=6）. The contents of 3 components in P. cuspidatum by above 2 methods had no significant difference. CONCLUSIONS： The method is simple, reasonable and reliable for quality control of P. cuspidatum.