Determination of Nicarbazin in Feeds Based on Mixed-Anionic Exchange Solid Phase Extraction by High Performance Liquid Chromatography-Tandem Mass Spectrometry

doi:10.11924/j.issn.1000-6850.casb2022-0187

Abstract

Abstract:

The objective of this study is to develop a method for the determination of nicarbazin in feeds based on mixed-anionic solid phase extraction (SPE) by high performance liquid chromatography - tandem mass spectrometric. The analyzed samples (compound feed, concentrated feed, and premixed feed) were extracted with methanol: acetonitrile (1:1), purified by mixed-anionic solid phase extraction column (MAX), dissolved in acetonitrile, separated on a C₁₈ chromatographic column (100 mm × 2.1mm, 2.6 μm) and performed with acetonitrile and ultrapure water as mobile phases, with the flow rate of 0.3 mL/min and the column temperature of 40℃ by gradient elution. The analysis was conducted by using negative-ion electrospray tandem mass spectrometry via multiple reactions monitoring (MRM) scan mode, and the samples were quantified by external standard method. The concentrations of nicarbazin in different feed matrix standard solutions had a good linear relationship in the range of 0.04-50 ng/mL, with correlation coefficients (r²) above 0.99, the limit of detection (LOD) was 0.02 mg/kg, and the limit of quantitation (LOQ) was 0.05 mg/kg. The recovery rate of nicarbazin with the content of 0.05-500 mg/kg in feed (0.05-10000 mg/kg in premixed feed) was 83.3%-105%, and the relative standard deviation (RSD) was 2.4%-7.6%. The method was sensitive, accurate, repeatable and suitable for the analysis of low and high levels of nicarbazin in compound feed, concentrated feed and premixed feed.

Key words: nicarbazin, LC-MS/MS, feed, anticoccidial drugs

WANG Weili, DING Chenhong, LIU Zhenni, QIN Shuzhen, YIN Qiumiao, LIN Xuexian, HUANG Xiaomei, WAN Kai, CUI Zefeng, LI Yafei. Determination of Nicarbazin in Feeds Based on Mixed-Anionic Exchange Solid Phase Extraction by High Performance Liquid Chromatography-Tandem Mass Spectrometry[J]. Chinese Agricultural Science Bulletin, 2023, 39(8): 133-141.

Figures/Tables 12

References 26

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目标物	母离子/(m/z)	产物离子/(m/z)	相对丰度比/%	Q1电压/V	碰撞能量/eV	Q3电压/V
尼卡巴嗪	300.90	137.05^a	100	15	13	13
尼卡巴嗪	300.90	107.05	5.94	15	37	10

目标物	母离子/(m/z)	产物离子/(m/z)	相对丰度比/%	Q1电压/V	碰撞能量/eV	Q3电压/V
尼卡巴嗪	300.90	137.05^a	100	15	13	13
尼卡巴嗪	300.90	107.05	5.94	15	37	10

饲料	回归方程	线性范围/(ng/mL)	相关系数	检测限LOD/(mg/kg)	定量限LOQ/(mg/kg)
配合饲料	y=310628x+36311.0	0.04~50	0.999	0.02	0.05
浓缩饲料	Y=316943x+93525.6	0.04~50	0.999
预混合饲料	y=289025x+78417.4	0.04~50	0.998

饲料	回归方程	线性范围/(ng/mL)	相关系数	检测限LOD/(mg/kg)	定量限LOQ/(mg/kg)
配合饲料	y=310628x+36311.0	0.04~50	0.999	0.02	0.05
浓缩饲料	Y=316943x+93525.6	0.04~50	0.999
预混合饲料	y=289025x+78417.4	0.04~50	0.998

饲料	添加浓度/ (mg/kg)	回收率/ %					平均回收率/ %	变异系数/ %
配合饲料	0.05	92.7	97.2	93.3	97.6	85.5	93.3	5.2
	1	83.7	87.2	87.5	91.9	94.6	89.0	4.8
	10	87.0	96.1	87.4	95.6	95.3	92.3	5.1
	100	92.2	88.8	89.5	96.3	95.8	92.5	3.7
	500	83.3	89.1	97.6	86.1	97.1	90.6	7.1
浓缩饲料	0.05	93.7	85.1	87.8	90.1	86.7	88.7	3.8
	1	94.0	93.7	86.7	94.5	86.6	91.1	4.5
	10	95.1	88.5	93.0	95.5	86.5	91.7	4.4
	100	90.6	94.3	87.7	92.7	85.2	90.1	4.1
	500	95.4	92.6	89.9	87.0	95.2	92.0	3.9
预混合饲料	0.05	98.0	92.9	98.2	101	90.8	96.2	4.4
	1	102	95.0	105	92.1	100	98.8	5.3
	10	101	95.0	94.9	104	103	99.6	4.4
	200	97.1	102	96.7	101	99.0	99.2	2.4
	10000	91.8	103	90.3	105	90.8	96.3	7.6