基于HPLC-ESI-MS/MS技术分析痰热清胶囊和痰热清注射液化学成分的差异

Comparison of Chemical Compositions Between Tanreqing Capsule and Tanreqing Injection by HPLC-ESI-MS/MS

目的:阐明痰热清的化学成分以及痰热清胶囊与痰热清注射液化学成分的差异。方法:采用高效液相色谱法串联质谱(HPLC-ESI-MS/MS)技术,乙腈-0.1%甲酸水为流动相进行梯度洗脱,电喷雾离子源负离子模式下采集数据,并通过准分子离子及二级碎片离子对化学成分进行鉴定。结果:痰热清胶囊中鉴定有机酸类成分7个、黄酮类成分10个、环烯醚萜类成分4个、木脂素类成分3个、苯乙醇苷类成分9个、胆汁酸类成分4个以及未知成分4个等共41个化合物。其中,经由对照品确证化学结构25个,推测化合物结构16个(含4个未知成分)。痰热清注射液中共鉴定化合物23个,相比胶囊注射液中连翘种苷异构体(3个),獐芽菜苷,连翘酯苷B,异连翘酯苷A,连翘酯苷A,芦丁,连翘种苷A异构体(2个),金丝桃苷,aldosecologanin,松脂素-β-D-葡萄糖苷/表松脂素-4'-O-葡萄糖苷/马台树脂醇-4'-O-葡萄糖苷异构体(2个),连翘苷,黄芩素,牛磺鹅去氧胆酸和黄芩提取物中所含的1个未知化合物等18个成分均缺失,可能是注射液无菌生产过程采用活性炭吸附、高温灭菌、配液等工艺以及过程控制中采用超滤技术,从而造成两种制剂化学成分的差异。结论:通过该研究建立的HPLC-ESIMS/MS分析方法,能有效表征痰热清胶囊和痰热清注射液的化学成分差异,可以为两种制剂的质量控制提供依据。

Objective： To identify the components in Tanreqing and clarify the differences between Tanreqing capsule and Tanreqing injection in chemical compositions. Method： An HPLC-ESI-MS/MS method was established and separation was performed with acetonitrile and 0. 1% formic acid as mobile phase for gradient elution. Mass spectrometric data were obtained in electrospray ionization （ESI） negative ion mode, and the chemical components were identified with quasi-molecular ions and secondary fragment ions. Result： Forty-one compounds including organic acids （7 compunds） , flavonoids （ 10 compounds）, iridoids （4 compunds） , lignans （3 compounds）, phenylethanoid glycosides （9 compounds）, bile acids （4 compounds） and 4 unknown components were detected in Tanreqing capsule by HPLC-ESI-MS/MS. Moreover, the chemical structures of 25 compounds were identified, and the chemical structures of 16 compounds （ including 4 unknown components） were inferred by their fragmentation patterns. However, only twenty three compounds were found in Tanreqing injection with the losses of 18 chemical compositions including three isolates of suspensaside, sweroside, forsythoside B, isoforsythiaside A, forsythiaside A, rntin, two isolates of suspensaside A, hyperin, aldosecologanin, two isolates of pinoresinol-/3-D-glucoside/epipinoresinol-4 ＇-O-glucoside/matairesinol-4 ＇-O-glucoside, phillyrin, baicalein, taurochenodeoxycholic acid （TCDCA） and one unknown compound derived from Scutellariae Radix extract. These differences in compositions between the two preparations may be caused by active carbon adsorption, high temperature sterilization, and blending system in aseptic production process and ultrafiltration technique in process control of Tanreqing injection product. Conclusion： The different chemical constituents in Tanreqing capsule and injection were characterized by the established HPLC-ESI-MS/MS method, which could provide the basis for the quality control of two preparations.