毒蘑菇中鹅膏肽类毒素的超高效液相色谱-四级杆飞行时间串联质谱同时测定法  

Simultaneous determination of amanita peptide toxins in poisonous mushrooms by ultra performanceliquid chromatography-mass spectrometer detector quadrupole time of flight

作　　者：黄小芹 陈汝梅 阮燕明 孙明月 林佶[3] HUANG Xiaoqin;CHEN Rumei;RUAN Yanming;SUN Mingyue;LIN Ji(School of Public Health,Kunming Medical University,Kunming Yunnan,650500,China;School of Public Health,Dali University,Dali Yunnan,671000,China;Physical and Chemical Inspection Center,Yunnan Center for Disease Control and Prevention,Kunming Yunnan,650032,China)

机构地区：[1]昆明医科大学公共卫生学院,云南昆明650500 [2]大理大学公共卫生学院,云南大理671000 [3]云南省疾病预防控制中心理化检验中心,云南昆明650032

出　　处：《职业与健康》2023年第17期2337-2344,共8页Occupation and Health

摘　　要：目的建立超高效液相色谱-四级杆飞行时间串联质谱法(ultra performance liquid chromatography-mass spectrometer detector quadrupole time of flight,UPLC-Q-TOF)定性定量检测毒蘑菇中鹅膏肽类毒素的分析方法。方法样品加入甲醇后均质,超声离心提取,待测液采用ACQUITY UPLC?HSS T_(3)1.8μm色谱柱(2.1×50 mm)进行分离,以0.1%氨水溶液-甲醇为流动相进行梯度洗脱0 min,2%B;0~0.5 min,2%B;0.5~6.0 min,95%B;6.0~7.0 min,95%B;7.0~8.0 min,2%B,流速0.3 mL/min,柱温40℃,进样量2.0μL。采用超高效液相色谱四级杆飞行时间质谱仪对毒蘑菇进行分离、鉴定鹅膏菌中的毒素成分。结果毒蘑菇样品中1、2、3、5、6、7、8号样品均未检出鹅膏肽类毒素;4号样品共检出6种组分,经鉴定为已知的鹅膏肽类毒素,分别是α-鹅膏毒肽、β-鹅膏毒肽、γ-鹅膏毒肽、二羟鬼笔毒肽、羧基二羟鬼笔毒肽和二羟鹅膏毒肽酰胺。进一步建立鹅膏肽类毒素快速定量方法,5种鹅膏肽类毒素在25~1000μg/L范围内呈现良好的线性关系,相关系数为0.9912~0.9989,平均加标回收率为89.5%~110.5%,相对标准偏差为3.80%~7.80%。其中α-鹅膏毒肽、β-鹅膏毒肽、γ-鹅膏毒肽、二羟鬼笔毒肽、羧基二羟鬼笔毒肽的含量分别是(19.701±0.175)、(0.098±0.014)、(1.548±0.001)、(0.331±0.002)及(0.108±0.060)mg/kg,二羟鹅膏毒肽酰胺与羧基二羟鬼笔毒肽相对含量为0.754 mg/kg。结论该方法用于测定毒蘑菇中多种鹅膏肽类毒素,方法简单、快速、可靠,为进食毒蘑菇突发中毒患者的快速临床诊断和及时治疗提供了有效的科学依据。Objective Establish an analytical method for the qualitative and quantitative detection of Amanita peptide toxins in poisonous mushrooms using ultra performance liquid chromatography mass spectrometer detector quadrupole time of flight(UPLCQ-TOF).Method The samples were homogenized with methanol and extracted by ultrasonic centrifugation.The liquid to be measured was separated by ACQUITY UPLC HSS T_(3)1.8μm column(2.1×50 mm),and gradient elution with 0.1%ammonia solution-methanol(0 min,2%B;0-0.5 min,2%B;0.5-6.0 min,95%B;6.0-7.0 min,95%B;7.0-8.0 min,2%B),with flow rate of 0.3 mL/min,column temperature of 40℃,and sample intake of 2.0μL.UPLC-Q-TOF was used to isolate and identify the toxin components of poisonous mushrooms.Result No amanita toxins were detected in samples 1,2,3,5,6 and 7 of the toxic mushroom samples.In sample 4,α-amanitin、β-amanitin、γ-amanitin、phallacidin、phalloidin and amanullin were detected.Further establish the rapid quantification method of Amanita toxins,5 kinds of Amanita toxins showed a good linear relationship within 25-1000μg/L,the correlation coefficient was 0.9912-0.9989,the average standard recovery rate was 89.5%-110.5%,the relative standard deviation(RSD)was 3.80%-7.80%.The content ofα-amanitin、β-amanitin、γ-amanitin、phalloidin and phallacidin was(19.701±0.175),(0.098±0.014),(1.548±0.001),(0.331±0.002)and(0.108±0.060)mg/kg respectively,and the relative content of amanullin and phallacidin was 0.754 mg/kg.Conclusion This method is simple,fast,and reliable for the determination of various amanita peptide toxins in poisonous mushrooms.It provides an effective scientific basis for the rapid clinical diagnosis and timely treatment of patients with sudden poisoning caused by eating poisonous mushrooms.

关 键 词：超高效液相色谱-四级杆飞行时间串联质谱法 鹅膏肽类毒素 野生菌 食物中毒 

分 类 号：R115[医药卫生—公共卫生与预防医学]