Mao Kun-jun, Huang Ping, Li Fu-qiang, Li Xiang, Chen Jian-wei
College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, PR China.
College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, PR China
J Chromatogr Sci. 2015 Oct;53(9):1449-54. doi: 10.1093/chromsci/bmv033. Epub 2015 Apr 1.
Hulisan tablets (HLST), a famous classic traditional Chinese prescriptions consisting of four medicinal herbs, have been applied for treating fractures, rheumatoid arthritis, mechanical phlebitis and traumatic bleeding extensively. In this study, a simple and reliable method using high-performance liquid chromatography with diode array detector (HPLC-DAD) was established for the simultaneous determination of 10 active compounds in HLST. The chromatographic analysis was performed on a Symmetry(®)-C18 column (4.6 × 250 mm, i.d., 5 µm; Waters, Wexford, Leinster, Ireland) at 30°C with a gradient elution of 0.1 mol/L ammonium acetate (containing 0.5 mL glacial acetic acid per 1,000 mL) and acetonitrile at a flow rate of 1.0 mL/min. The detection UV wavelengths were set at 232, 254 and 280 nm. The method was validated by linearity, precision, stability and recovery. Calibration curves for the 10 compounds showed good linear regressions (R(2) > 0.9992). The limits of detection and quantification fell in the ranges of 0.03-0.36 and 0.11-1.01 µg/mL, respectively. The results of the recovery test were 97.19-102.04% with a RSD value of 0.65-2.47%. The developed method was subsequently applied to evaluate five batches of HLST and testified to be suitable for the quality control.
虎力散片（HLST）是一种由四味药材组成的著名经典中药方剂，已广泛应用于治疗骨折、类风湿性关节炎、机械性静脉炎和外伤性出血。本研究建立了一种简单可靠的高效液相色谱-二极管阵列检测器（HPLC-DAD）法，用于同时测定虎力散片中的10种活性成分。色谱分析在Symmetry(®)-C18柱（4.6×250 mm，内径，5 µm；沃特世公司，韦克斯福德，伦斯特，爱尔兰）上进行，柱温30℃，以0.1 mol/L醋酸铵（每1000 mL含0.5 mL冰醋酸）和乙腈进行梯度洗脱，流速为1.0 mL/min。检测紫外波长设定为232、254和280 nm。该方法通过线性、精密度、稳定性和回收率进行验证。10种化合物的校准曲线显示出良好的线性回归（R(2)>0.9992）。检测限和定量限分别在0.03 - 0.36和0.11 - 1.01 µg/mL范围内。回收率试验结果为97.19 - 102.04%，相对标准偏差值为0.65 - 2.47%。所建立的方法随后应用于评价五批虎力散片，并证明适用于质量控制。
