HPLC法同时测定清肝散结方中8种化学成分的含量

Simultaneous Determination of 8 Chemical Constituents in Qinggan Sanjie Formula by HPLC

目的：建立同时测定清肝散结方中岩白菜素、绿原酸、黄芩苷、汉黄芩苷、野黄芩苷、黄芩素、汉黄芩素及芹菜素含量的方法。方法：采用高效液相色谱法。色谱柱为Phenomenex Gemini C18（150 mm×3 mm,3μm）,流动相为0.2%甲酸-0.2%甲酸甲醇（梯度洗脱）,流速为0.4 ml/min,柱温为20℃,检测波长为270 nm,进样量为4.0μl。结果：岩白菜素、绿原酸、黄芩苷、汉黄芩苷、野黄芩苷、黄芩素、汉黄芩素及芹菜素的质量浓度分别在5.00～199.80、5.01～200.40、20.04～801.60、3.71～148.32、3.46～138.60、4.23～169.12、4.59～183.60、2.86～114.24μg/ml范围内与各自峰面积积分值呈良好线性关系,r均为0.999 9;精密度试验的RSD〈1%,稳定性、重复性试验的RSD〈3%;平均加样回收率分别为101.23%（RSD=0.52%）、100.69%（RSD=0.75%）、100.89%（RSD=1.18%）、100.21%（RSD=0.80%）、99.26%（RSD=1.33%）、100.09%（RSD=1.78%）、100.93%（RSD=1.73%）、100.33%（RSD=1.20%）,n均为6。结论：该方法结果准确可靠、重复性好,实用性强,可用于清肝散结方中8种化学成分含量的测定。

OBJECTIVE： To establish a method for simultaneous determination of bergeninum, chlorogenic acid, baicalin, wogonoside, scutellarin, baicalein, wogonin and apigenin in Qinggan sanjie formula. METHODS： HPLC method was adopted. The determination was performed on Phenomenex Gemini C18（ 150 mm×3 mm, 3 μm） with mobile phase consisted of 0.2% formic acid-0.2% formic acid methanol （gradient elution） at the flow rate of 0.4 ml/min. The column temperature was 20 ℃, and the de- tection wavelength was set 270 nm. The sample size was 4.0 μl. RESULTS： The linear range were 5.00-199.80 μg/ml for bergeni- num（r=0.999 9）, 5.01-200.40 μg/ml for chlorogenic acid（r=0.999 9）, 20.04-801.60 μg/ml for baicalin（r=0.999 9）, 3.71-148.32 μg/ml for wogonoside（r=0.999 9）, 3.46-138.60 μg/ml for scutellarin（r=0.999 9）,4.23-169.12μg/ml for baicalein（r=0.999 9）, 4.59-183.60 μg/ml for wogonin （r=0.999 9）and 2.86-114.24 μg/ml for apigenin （r=0.999 9）, respectively. RSDs of precision test was lower than 1%, and RSDs of stability and reproducibility tests were lower than 3%. The average recoveries were 101.23% （RSD=0.52% , n=6）, 100.69% （RSD=0.75%, n=6）, 100.89% （RSD= 1.18%, n=6） , 100.21% （RSD=0.80% , n=6） , 99.26% （RSD= 1.33%, n=6）, 100.09% （RSD= 1.78%, n=6）, 100.93% （RSD= 1.73%, n=6）and 100.33% （RSD= 1.20%, n=6）. CON- CLUSIONS ： The method is accurate and reproducible, and can be used for the quality control of Qingan sanjie formula.