China Animal Husbandry and Veterinary Medicine ›› 2022, Vol. 49 ›› Issue (7): 2746-2756.doi: 10.16431/j.cnki.1671-7236.2022.07.033

• Basic Veterinary Medicine • Previous Articles     Next Articles

Methodology Study on Determination of Diflubenzuron in Swine Plasma by Liquid Chromatography-tandem Mass Spectrometry

XU Yuzhen1,2, LIU Xiwang2, QIN Zhe2, LI Shihong2, BAI Lixia2, FAN Liping2, WEI Yanming1, LI Jianyong1,2, YANG Yajun2   

  1. 1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
    2. Key Laboratory of New Animal Drug Project of Gansu Province, Key Laboratory of Veterinary Pharmaceutical Development of Ministry of Agriculture and Rural Affairs, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences, Lanzhou 730050, China
  • Received:2022-02-11 Online:2022-07-05 Published:2022-06-29

Abstract: 【Objective】 To investigate the pharmacokinetics character of the diflubenzuron (DFB) premix in swine after drenching administration,a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to determine diflubenzuron in swine plasma.【Method】 DFB-13C6 was employed as internal standard.Plasma samples were processed by protein precipitation of methanol.After centrifugation,the supernatant was vacuum evaporation to dry,and the residue was resolved by the solvent of methanol-water (1∶1).The column of ZORBAX Eclipse Plus C18 was used to separate the analyte.Mobile phase was methanol-buffer of 0.1% formic acid and 5 mmol/L ammonium formate with flow rate of 0.4 mL/min under gradient elution.The column temperature was 35 ℃ and injection volume was 10 μL.Determination of DFB was performed using LC-MS/MS with an electrospray ionization source interface operated in the positive-ion scan mode under the multiple reaction monitoring mode.Product ions 311.1→158.1 and 317.1→158.1 were employed as quantifier ion of DFB and DFB-13C6,respectively.The conditions of LC and MS were optimized.The proposed method was validated through specificity,matrix effects,quantitative lower limits,lineral range,precision,accuracy and samples stability.【Result】 The established method had high sensitivity,the limit of detection (LOD) was 0.5 ng/mL and the limit of quantitative (LOQ) was 1 ng/mL.Matrix effects had little influence on the detection of samples.The standard solution had a good linear relationship in the concentration range of 1-1 000 ng/mL (R2≥0.998).The average accuracy was during 97.78% to 115.06% and the intra-assay and inter-assay precisions were both less than 10% when the spiked levels of DFB in plasma were LOQ (1 ng/mL),low (2 ng/mL),medium (100 ng/mL) and high concentration (800 ng/mL).These results could meet the methodological requirements.The standard solution,spiked samples and processed samples had good stability when they stored at the specific conditions except freeze-thaw cycles.【Conclusion】 The established LC-MS/MS detection method was simple,specific,sensitive,accurate and reliable.It could be used to detect the concentration of diflubenzuron in pig plasma,and then applied to the pharmacokinetics of diflubenzuron premix in swine.

Key words: swine; plasma; diflubenzuron (DFB); LC-MS/MS; internal standard method

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