China Animal Husbandry and Veterinary Medicine ›› 2022, Vol. 49 ›› Issue (12): 4843-4853.doi: 10.16431/j.cnki.1671-7236.2022.12.032

• Basic Veterinary Medicine • Previous Articles     Next Articles

Simultaneous Determination of 4 Isoquinoline Alkaloids in Broiler Tissues by Ultra-performance Liquid Chromatography-tandem Mass Spectrometry

LIU Mengting1,2, HU Nanxi1,3, ZOU Xianglin1,2, YANG Zihui1,2, ZENG Jianguo1,2   

  1. 1. Hunan Key Laboratory of Traditional Chinese Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China;
    2. College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China;
    3. Hunan Puruima Drug Research Center Co., Ltd., Changsha 410324, China
  • Received:2022-08-01 Online:2022-12-05 Published:2022-12-01

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Abstract: 【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-C18, 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.

Key words: isoquinoline alkaloids; UPLC-MS/MS; trace analysis

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