基于HPLC-Q TOF-MS/MS技术分析纯化后菟丝子拟雌激素活性成分

Analysis of Purified Active Ingredients of Estrogen Mimic Effect in Cuscuta chinensis by HPLC-Q TOFMS /MS

目的探讨菟丝子中拟雌激素活性成分的分析方法,为建立高效液相色谱-四级杆飞行时间串联质谱法提供依据。方法采用D-101型大孔吸附树脂纯化菟丝子拟雌激素活性成分,并结合HPLC-Q TOF-MS/MS技术对纯化后的菟丝子拟雌激素活性成分进行分析。结果建立了高效液相色谱-四级杆飞行时间串联质谱方法,对经由D-101型大孔吸附树纯化后的菟丝子进行成分分析,共推测出13个活性成分。分别考察了洗脱液的浓度、pH值、体积,最终选择体积分数70%乙醇洗脱为最佳洗脱液。已推测出的13个成分分别为槲皮素-3-O-(2″-O-α-鼠李糖-6″-O-丙二酰基)-β-D-葡糖苷,山柰酚-3-O-β-D-芹菜糖-(1→2)-[-α-L-鼠李糖-(1→6)]-β-D-葡糖苷,6-O-(反式)对香豆酰基-β-D-呋喃果糖-(2→1)-α-D-吡喃葡萄糖苷,山柰酚-7-鼠李糖苷,山柰酚-3-β-D-葡萄糖醛酸苷,芹菜素,山柰酚-3-呋喃阿拉伯糖苷,槲皮素-3-O-β-D-半乳糖-(2→1)-β-D-芹糖苷,二咖啡酰奎尼酸,金丝桃苷,槲皮苷,绿原酸,槲皮素。结论菟丝子中含有拟雌激素作用的活性成分。

OBJECTIVE To establish a high-performance liquid chromatography-quadrupole time of flight tandem mass spectrom- etry （ HPLC-Q TOF-MS/MS） method for analysis of estrogen-like active ingredients in Cuscuta chinensis. METHODS D - 101 macroporous resin was used for purification of cuscuta chinensis estrogen-like active ingredients, and HPLC-Q TOF-MS/MS technology was used to analyze the purified estrogenic active ingredients in cuscuta ehinensis. The concentration, pH and volume of the eluent were investigated, and uhimately 70% ethanol was selected. RESULTS An HPLC-Q TOF-MS/MS method was developed. A total of 13 active ingredients were inferred, which were quercetin-3-O-（2＂-O-a-rhamnose-6＂-O-malonyl）-β-D-glueoside, kaempferol-3-O-β-D- apiose- （ 1 →2 ） - [ -α-L-rhamnose- （ 1 →6 ） ] -/3-D-glucoside, 6-0- （ E ） -P-coumaroyl- β-D-fructofuranosyl- （ 2 → 1 ） -α-D-glucopyranoside, kaempferol-7-rhamnoside, kaempferol-3-β-D-glucuronide, apigenin, kaempferol-3-furan arabinoside, quercetin-3-O-β-D-galactose-（ 2→1 ）-β-D-glacoside celery, dicaffeoylquinic acid, hyperoside, quercitrin, chlorogenic acid and quercetin. CONCLUSION Cuscuta chinensis contain active ingredients of estrogen mimic effect.