摘要
【目的】试验旨在建立一种同时测定肉鸡组织中原阿片碱、别隐品碱、白屈菜红碱和血根碱含量的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法,以便更好地监测博落回散和博普总碱散活性成分在肉鸡组织中的分布与残留情况。【方法】对肉鸡组织进行前处理方法优化,以0.1%甲酸水和乙腈为流动相,在Agilent Zorbax SB-C18上梯度洗脱,流速为0.3 mL/min,柱温为35℃,进样量为1μL,在正离子模式下通过多反应监测模式采集数据,采用内标校正和外标法测定生物碱含量,并对方法线性、检测限与定量限、基质效应、回收率、批内与批间精密度、准确度及稳定性进行考察。【结果】白屈菜红碱、原阿片碱和别隐品碱的线性范围为0.5~200μg/L,血根碱为2~200μg/L,相关系数均≥0.999,线性关系良好;白屈菜红碱、原阿片碱和别隐品碱的检测限与定量限分别为0.2和0.5μg/L,血根碱检测限与定量限分别为1和2μg/L。4种生物碱在高、中、低浓度下方法回收率达到86.61%~111.17%,批内与批间精密度和准确度均≤14.93%,基质效应为81.73%~120.78%,稳定性的相对标准偏差均≤13.55%。【结论】本试验建立了同时检测肉鸡心脏、肝脏、肾脏、胸肌和腿肌中4种异喹啉类生物碱的超高效液相色谱-串联质谱法,方法符合各项方法学考察要求,具有专属性强、灵敏度高、方便快捷等特点,可实现对肉鸡组织中痕量生物碱的精确、快速定量。
【Objective】 The purpose of the experiment was to establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of protopine, allocryptopine, chelerythrine and sanguinarine in broiler tissues, so as to better monitor the distribution and residue of active ingredients of Boluohui powder and Bopuzongjian powder in broiler tissues.【Method】 The pretreatment method of broiler tissues was optimized, with 0.1% formic acid water and acetonitrile as the mobile phase, gradient elution on Agilent Zorbax SB-C_(18),the flow rate was 0.3 mL/min, the column temperature was 35 ℃,and the injection volume was 1 μL,multiple reaction monitoring mode for data acquisition in positive ion mode, the content was calculated by the external standard method, corrected by the internal standard, and the linearity, limit of detection and limit of quantification, recovery, intra-and inter-run precision, accuracy and stability of the method were investigated.【Result】 The results showed that the linear range of chelerythrine, protopine and allocryptopine were 0.5-200 μg/L,sanguinarine was 2-200 μg/L,the linear relationship was good, and the correlation coefficients were greater than 0.999.The detection and quantification limit of chelerythrine, protopine and allocryptopine were 0.2 and 0.5 μg/L respectively, and sanguinarine were 1 and 2 μg/L respectively.The recovery rate of the four alkaloids were 86.61%-111.17% at high, middle and low concentrations, the precision and accuracy were ≤14.93% within and between batches, and the matrix effect was 81.73%-120.78%,RSD of stability were all less than 13.55%.【Conclusion】 The established ultra-performance liquid chromatography-tandem mass spectrometry method for simultaneous detection of four isoquinoline alkaloids in the heart, liver, kidney, chicken breast and chicken thigh of broilers was consistent with various methodological investigations, and it was specificity, sensitivity, convenience and rapidness.It was suitable for accurate and rapid quantification of trace alkaloids in broiler tissues.
作者
刘梦婷
胡南希
邹湘霖
杨子辉
曾建国
LIU Mengting;HU Nanxi;ZOU Xianglin;YANG Zihui;ZENG Jianguo(Hunan Key Laboratory of Traditional Chinese Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China;College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China;Hunan Puruima Drug Research Center Co.,Ltd.,Changsha 410324,China)
出处
《中国畜牧兽医》
CAS
北大核心
2022年第12期1743-1753,共11页
China Animal Husbandry & Veterinary Medicine
基金
湖南省研究生科研创新项目(QL20210162)。
