加速溶剂萃取法结合HPLC-ESI-TOF MS测定清肝散结汤中29种化学成分（英文）

Determination of 29 Chemical Constituents in the QingGanSanJie Decoction by Accelerated Solvent Extraction and HPLC-ESI-TOF MS

清肝散结汤(QGSJ)是由14味不同中药组成的复方制剂,基于长期的实验观察和临床经验以不同组合成方用于肝癌的治疗。本研究建立了一种基于加速溶剂萃取(ASE)结合高效液相色谱-电喷雾飞行时间质谱(HPLC-ESI-TOF MS)的方法同时测定清肝散结汤中29种化学成分。使用70%乙醇作为提取溶剂,提取温度100℃,静态萃取时间5min,提取2次,以上条件采用正交设计和主成分分析进行优化。HPLC分离采用Kromasil C18色谱柱,梯度洗脱。TOF MS在负离子模式下检测,质量扫描范围m/z100~1 100。结果表明:清肝散结汤中29种成分具有良好的线性关系(r>0.994)和日内、日间精密度(RSD<5%);提取回收率在96.5%~104.5%之间。该方法快速、可靠,适用于复方中药的定量评价。

QingGanSanJie decoction （QGSJ）, which consists of 14 different herbs, is a multiherbal formula for liver cancer that is administered in different combinations based on experimental observations and clinical experiences from long-term applications. A method based on accelerated solvent extraction （ASE） followed by high-performance liquid chromatography （HPLC） coupled with electrospray time-of-flight mass spectrometry （ESI-TOF MS） were established for simultaneous determination of 29 chemical components in QGSJ. The operational parameters of ASE, including the extraction solvent （70% ethanol）, extraction temperature （100 ℃）, static extraction time （5 min）, and extraction cycles （two cycles） were optimized by orthogonal design and principal component analysis. HPLC separation was performed on a Kromasil C18 HPLC column with a linear gradient mobile phase system. TOF MS was set in negative ion mode with a detector range of m/z100-1100. The results show that 29 compounds in the QGSJ have good linearity （r〉0.994）, and intra-day and inter-day precision （relative standard deviation） are less than 5%. The extraction recoveries of the 29 compounds range from 96.5% to 104.5%. The method is rapid and reliable, which is suitable for the quantitative evaluation of multiherbal traditional Chinese medicine.