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通脉颗粒是由丹参、川芎、葛根三味中药的水提物制成的用于治疗心血管疾病的复方制剂,主要成分为异黄酮苷、异黄酮及酚酸类.低极性化合物如丹参酮和酚酞类也可能对活性有贡献,但很难检测到.因此,本研究建立了正向×反相离线二维液相色谱方法对通脉颗粒中的有效成分进行富集和分离.采用正向SEP-PAK硅胶固相萃取柱分离通脉颗粒的乙酸乙酯萃取物,分别以石油醚、乙酸乙酯-甲醇(6∶1,v/v)、甲醇洗脱获得三个流分,然后采用超高效液相反向色谱串联四级杆-静电场轨道阱质谱对其进行检测.葛根中黄酮类成分,丹参中酚酸类成分及川芎中酚酞类成分分布于不同流分中.从通脉颗粒中初步鉴定了131个化合物,其中葛根59个,丹参26个,川芎43个,丹参和川芎的共有成分3个.26个化合物经过对照品比对鉴定.本研究为中药复方成分的全面分析提供了有效方法.“,”TongMai Granules is a Chinese medicine patent drug to treat cardiovascular diseases.It is prepared from the water extract of Dan-shen,Chuan-xiong and Ge-gen.The main compounds of TongMai Granules are isoflavone glycosides,isoflavones,and phenolic acids.However,it is difficult to detect low-polar compounds such as tanshinones and phthalides though they may also contribute to the therapeutic effect.In this work,an off-line two-dimensional chromatography method was developed to enrich the minor constituents in TongMai Granules.The extract was subjected to an SEP-PAK silica SPE column and eluted with petroleum ether,ethyl acetate-methanol(6∶1,v/v),and methanol,successively.The three fractions were then analyzed using ultra-high performance liquid chromatography coupled with orbitrap mass spectrometry(UHPLC/orbitrap-MS).The isoflavones from Ge-gen,phenolic acids from Dan-shen and phthalides from Chuan-xiong were distributed in different fractions.A total of 131 compounds,including 59 from Ge-gen,26 from Dan-shen,43 from Chuan-xiong,and 3 common ones from Dan-shen and Chuan-xiong were tentatively identified.Among them,26 compounds were identified by comparing with reference standards.This study set a good example for comprehensive analysis of the constituents in traditional Chinese medicine formula.