基于UPLC-Q-Extractive Orbitrap MS鉴定姜黄提取物在大鼠体内的代谢产物

Identification of Metabolites of Turmeric Extract in Rats by UPLC-Q-Extractive Orbitrap MS

本研究基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Extractive Orbitrap MS),鉴定了姜黄提取物中姜黄素、去甲氧基姜黄素、双去甲氧基姜黄素3种姜黄素类化合物在大鼠体内的代谢产物。对健康雄性大鼠单剂量灌胃给予1703 mg/kg姜黄提取物(相当于480 mg/kg总姜黄素),收集粪便、尿液、胆汁、血浆生物样品,采用UPLC-Q-Extrative Orbitrap MS在负离子模式下鉴定代谢产物。在大鼠体内共鉴定出59个代谢物(B1~W12),其中B7、B8、B11~B13、C3、D3、D5、D8、D10、W1、W2、W4、W5、W7、W10为新发现的代谢产物,W3/W6、W11/W12、W12分别为血浆、粪便、尿液中首次发现的代谢产物。3种姜黄素类化合物在大鼠体内的代谢途径主要为葡萄糖醛酸化、硫酸酯化、还原氢化、氧化、还原、羟基化、去羟基化和多种反应类型的复合反应等。

Turmeric extract is aβ-diketone polyphenol extract isolated from the Curcuma longa L,a plant of the genus Curcuma of the Zingberaceae family.It mainly includes curcumin(CUR),demethoxycurcumin(DMC),and bisdemethoxycurcumin(BDMC).It is a commonly used food,natural pigment and cosmetic additive,which has various pharmacological activities,such as hypolipidemia,antioxidant,anti-inflammation,antiviral and anti-Alzheimer's disease.However,poor stability and low bioavailability limit the clinical application of curcuminoids.Curcumin hydrogenated metabolites also have pharmacological activity,and they are more stable and bioavailable than curcumin.The metabolites of curcuminoids(CUR,DMC and BDMC)in turmeric extract in rats were identified.Turmeric extract was suspended in 0.5%carboxymethlcellulose sodium(CMC-Na).Healthy male Sprague Dawley(SD)rats in drug group were gavaged with a single dose of 1703 mg/kg turmeric extract(approximately 480 mg/kg curcuminoids),and the rats in control group were given equal volume of 0.5%CMC-Na solution.And then,plasma at different time points was collected after oral gavage.Urine,faeces and bile were collected after oral gavage in rats within 12 h.Plasma were prepared by protein precipitation method,the urine,faeces and bile were extracted by methanol.The biological samples of faeces,urine,bile and plasma were analyzed by ultra high per-formance liquid chromatography-quadrupole extractive orbitrap mass spectrometry(UPLC-Q-Extractive MS)in negative ion mode.The accurate molecular weight,primary and secondary mass spectrometry information of metabolites in faeces,urine,bile and plasma were identified.As a result,a total of 59 metabolites(B1-W12)were identified.B3,B7,B8,B11,B12,B13,C3,D3,D5,D8,D10,W1,W2,W4,W5,W7 and W10 were new metabolites.W3/W6,W11/W12 and W12 were firstly found in plasma,faces and urine,respectively.The results showed that the main metabolic pathways of curcumin were glucuronidation,sulfation,reductive hydrogenation,oxidation,reduction,hydroxylation,dehydroxylation and their composite reactions.This method can provide a theoretical basis for the later screening of active metabolites of curcumin compounds.