高效液相色谱-三重四极杆质谱法同时测定烟叶中10种多酚类化合物

Simultaneous determination of ten polyphenols in tobacco leaves with high performance liquid chromatography-triple quadrupole mass spectrometry

为研究烟叶中的植物多酚类化合物,利用高效液相色谱-三重四极杆质谱(HPLC-TQMS)技术,建立了同时分析烟叶中山奈酚(Kaempferol)、花青素(Cyanidin)、氯化飞燕草色素(Delphinidin chloride)、鼠李金(Rhamnazin)、绿原酸(Chlorogenic acid)、氯化飞燕草素3-O-β-吡喃葡糖苷(Delphinidin3-O-β-glucopyranoside chloride)、原花青素B2(Procyanidine B2)、氯化花青素鼠李葡糖苷(Keracyaninchloride)、芸香苷(Rutin)和莨菪亭(Scopoletin)等10种植物多酚类化合物质量分数的方法。采用实时离子多反应检测模式,获得10种植物多酚类化合物的特征碎片离子峰,并对烟草花叶病毒(TMV)感染的烟叶样品进行了测定。结果表明:①10种植物多酚类化合物在线性范围内的相关系数(r)为0.999 0~0.999 6,方法的检出限为1~200μg/L,日内精密度(RSD)为0.64%~8.72%,日间精密度(RSD)为1.05%~7.93%;回收率为79.66%~112.74%;②TMV感染极显著(p<0.01)增加了烟叶中山奈酚、花青素、氯化飞燕草色素、氯化飞燕草素3-O-β-吡喃葡糖苷和芸香苷的质量分数,显著(p<0.05)降低了烟叶中鼠李金的质量分数,增加了绿原酸的质量分数;③TMV病毒感染对原花青素B2、氯化花青素鼠李葡糖苷和莨菪亭的影响不大,差异不显著。该方法具有良好的灵敏度、精密度和回收率,可用于同时分析植物多酚类化合物。

In order to study polyphenols in tobacco,a method for simultaneous determination of ten endogenous polyphenols in tobacco was developed with high performance liquid chromatography-triple quadrupole mass spectrometry(HPLC-TQMS).The ten polyphenols included kaempferol,cyanidin,delphinidin chloride,rhamnazin,chlorogenic acid,delphinidin 3-O-β-glucopyranoside chloride,procyanidine B2,keracyanin chloride,rutin,and scopoletin.The characteristic fragment ion peaks of the ten polyphenols were obtained by dynamic multiple reaction monitoring(DMRM).Tobacco mosaic virus(TMV)-infected tobacco samples were also included in the study.The results showed that:1)The linear correlation coefficients(r)of the ten polyphenols were from 0.999 0 to 0.999 6.The limits of detection(LODs)for all the analytes were 1-200μg/L,and the intra-day and inter-day relative standard deviations(RSDs)were 0.64%-8.72%and 1.05%-7.93%,respectively.The recoveries were in the range of 79.66%-112.74%.2)The contents of kaempferol,cyanidin,delphinidin chloride,delphinidin 3-O-β-glucopyranoside chloride and rutin in TMV-infected tobacco samples were significantly higher(p<0.01)than those in the control,and the rhamnazin content was significantly lower(p<0.05)and the chlorogenic acid content was higher in TMV-infected tobacco than in the control.3)The contents of procyanidine B2,keracyanin chloride,and scopoletin were not significantly affected by TMV infection.This method could be used to simultaneously determine a variety of endogenous polyphenols with good sensitivity,precision and recovery.