UPLC-MS/MS测定AM真菌侵染丹参植物脱落酸含量  

Determination of Abscisic Acid in Arbuscular Mycorrhizal Fungi infected Salvia Miltiorrhiza Plants by UPLC/MS/MS

作　　者：周修腾[1,2] 康利平[2] 李鹏英[2] 杨光[2] 王雪[2,3] 陈美兰[2] 

机构地区：[1]广州中医药大学临床药理研究所,广州510405 [2]中国中医科学院中药资源中心,道地药材国家重点实验室培育基地,北京100700 [3]北京中医药大学东方医院药学部,北京100078

出　　处：《中国农学通报》2016年第12期92-97,共6页Chinese Agricultural Science Bulletin

基　　金：国家中医药局行业科研专项经费“30项中药材生产使用技术规范化及其适用性研究”(201407005)

摘　　要：旨在建立一种分析丹参植物体内脱落酸含量的方法,揭示AM真菌对丹参脱落酸含量的影响。本研究采用超高效液相色谱-三重四级杆质谱（UPLC-MS/MS）,植物激素采用异丙醇：水：HCI（2：1：0.002）提取液提取,二氯甲烷萃取,进而应用超高效液相色谱-质谱联用,以多反应监测（MRM）进行定量分析。脱落酸在线性范围内的相关系数（r）为0.9986;检出限为0.0466 ng/m L;植物根和叶片高、中、低3个浓度加标回收率在80.48%～96.41%之间;日内和日间精密度RSD均低于10.0%;稳定性和重复性RSD均低于5.0%,接种AM真菌丹参植物根和叶片中脱落酸含量均显著低于对照组。该方法具有良好的灵敏度、精密度和回收率,适用于丹参植物内源激素脱落酸的含量检测。The study aims to establish a method analyzing the content of endogenous abscisic acid in salvia miltiorrhiza, and reveal the effect of arbuscular mycorrhizal（AM） on the content of abscisic acid. The abscisicacid analysis was conducted using ultra- high performance liquid chromatography- triple quadrupole massspectrometry（UPLC-MS/MS）. The phytohormone was extracted with 2-propanol： H2O： concentrated HCl（2：1：0.002, v/v/v）, extracted by dichloromethane, and determined using reverse- phase liquid chromatography-tandem mass spectrometry, and the quantitative analysis was carried out with multiple reaction monitoring. Inthe linear ranges, the correlation coefficient（r） of abscisic acid was 0.9986. The detection limits was 0.0466 ng/m L. The recovery rates of high, medium and low concentration were 80.48%-96.41% in root and leaf. Therelative standard deviations（RSD） of inter-day precision and within-day precision were under 10.0%. AndRSD values of stability and reproducibility were lower than 5.0%. After inoculated with AM fungi, the contentof abscisic acid in root and leaf were significantly lower than the control group. This method had goodsensitivity, precision, recovery, and could be used to determine the content of abscisic acid in salvia miltiorrhiza.

关 键 词：脱落酸 超高效液相色谱-三重四级杆质谱 丹参 丛枝菌根真菌 

分 类 号：S131.2[农业科学—农业基础科学]