5 11 Vol. 5 No. 11 2014 11 Journal of Food Safety and Quality Nov., 2014 HPLC-MS/MS 张 军 1*, 杜平 2 (1., 831100; 2., 830002) 摘要 : 目的 - 方法 40% (v:v), 0.22 μm, Agilent ZORBAX RX-SIL (100 mm 3.0 mm,1.8 μm) ; 0.2% +10 mmol/l A, B ; 0.4 ml/min (ESI + ), (MRM) 结果 0.2~0.5 μg/l 1.0 5.0 10.0 mg/kg 92.2%~98.7%, 1.9%~6.7% 结论, 关键词 : ; ; ; Optimized detection method for chlormequat and mepiquat in cotton seed oil and its by-products using high performance liquid chromatography-mass spectrometry ZHANG Jun 1*, DU Ping 2 (1. Xinjiang Agricultural Vocational Technical College, Changji 831100, China; 2. Xinjiang Chalkis Natural Plant High-Tech Institute, Urumqi 830088, China) ABSTRACT: Objective To establish a detection method of chlormequat and mepiquat from cottonseed oil and by-products by high performance liquid chromatography-mass spectrometry (HPLC-MS/MS). Methods The samples were extracted by water or containing 40% acetonitrile (v:v) buffer solution, and purified by 0.22 μm filter membrane, and separated in Agilent ZORBAX RX-SIL column (100 mm 3.0 mm, 1.8 μm). The mobile phase was divided into A solution with 0.2% formic acid +10 mmol/l ammonium acetate aqueous, B solution with methanol, to elute of a flow rate at 0.4mL/min. Electrospray ionization in the positive ion mode (ESI + ), multiple reaction monitoring mode (MRM) detection. Results It showed there was a good linearity in the range of 0.2~0.5 μg/l for chlormequat and mepiquat, the recovery of standard addition was 92.2%~98.7%, the relative standard deviation was 1.9~6.7% in 1, 5, and 10.0mg/kg levels. Conclusion The method could better meet the test requirement in sensitive, accurate, and the recovery rate. KEY WORDS: cottonseed oil; cottonseed meal; chlormequat; mepiquat 基金项目 : (XJNZYKJ2012008) Fund: Supported by Xinjiang Agricultural Vocational Technical College Natural Science Key Project (XJNZYKJ2012008) * 通讯作者 :,, E-mail: wusuzj@163.com *Corresponding author: ZHANG Jun, Associate Professor, Xinjiang Agricultural Vocational Technical College, No. 29, East Cultural Road, Changji 831100, China. E-mail: wusuzj@163.com
11, : HPLC-MS/MS 3685 1 引言 (chlormequat),,, : 2-,,,,,,,, [1] (mepiquat), 鎓,, : 1, 1-,,,,,,,,,,,,, [2-5] [6], [7],,, [8], [9], (), (CAC) 50, 136, 130 [10], GB 1537-2003,,, GB/T5009.37-2003, ( ), [11],,, - [12-13] - [14-16],,, (), - (HPLC-MS/MS),,, 2 材料与方法 2.1 仪器与试剂 Agilent RRLC1200,Triple Quad MS/MS6410B - (); BS210S ( ); HR1617(); GT10-1( ); KQ5200DB( ) (, TEDIA ); ( >99%, Dr.Ehrenstorfer ); (, ); ; () 2.2 溶液的配制 2.2.1 标准溶液的配制 : 10.0 mg 1 100 ml,, 100.0 mg/l :, 2.5 0.25 0.05 mg/l : 4,, 1.0 2.0 5.0 10.0 20.0 50.0 μg/l 2.2.2 缓冲溶液配制 3.85 g,, 2 g
3686 5, 1 L 2.3 样品前处理方法 : 3.0 g 10 ml, -(2:3,v:v) 4 ml, 3 ml,, 3 ml 3,, 5 ml, 0.22 μm : 20, 3 2, 30 min, 3.0 g 50 ml, 40% (v:v) 30 ml, 1 h 1 min, 10 min, 9000 r/min 6 min, 0.22 μm :, 3.0 g 50 ml, 30 ml, 1 min, 10 min, 9000 r/min 6 min, 0.5 ml, 40%(v:v) 25 ml, 30 min, 0.22 μm : 1 4, 2.4 实验条件 2.4.1 色谱条件 : Agilent ZORBAX RX-SIL (100 mm 3.0 mm,1.8 μm); : A 0.2% +10 mmol/l, B ; : 0.4 ml/min; : 40 ; : 5 μl 2.4.2 质谱条件 (ESI + ) ; 4000 V; 350 ; 10 L/min; 276 kpa 1 3 结果与讨论 3.1 样品前处理的优化,,,,, 3 : - (2:3, v:v)- (2:3, v:v),, ( 2), -, -,,,,, - (- ),,,,,,,, - - -( 2:3, v:v), :, -;,, Table 1 表 1 矮壮素 缩节胺的 MS/MS 采集参数 Chlormequat and Mepiquat chloride MS/MS analysis parameters Fragmentor/V Precursorion(m/z) Production(m/z) Collision energy/v Chlormequat () 120 122 58* 30 Mepiquat chloride () 63 20 120 114 98* 30 58 30 *quantitative production
11, : HPLC-MS/MS 3687 Table 2 表 2 不同提取溶剂的提取效果比较 ( 峰面积 ) Comparison of the extraction effect from different extraction solvent (peak area) 14316 4752 5230 318 1586 37235 18189 6340 12437 952 1654 37593-18591 6749 27699 3075 1695 37706-18013 6712 20594 2024 1636 37719 1 10.0 μmol/l 4 (1.; 2.) a.10.0 μmol/l ; b.; c. ; d.; e. Fig. 1 Total ion chromatogram of 10.0 μmol/l mixed standard solution and 4 samples in chlormequat and mepiquat (1.chlormequat 2. mepiquat) a.10.0 ppb mixed standard solution of chlormequat and mepiquat; b.cotton seed hulls; c.cottonseed meal; d.cottonseed oil; e.mushroom Table 3 表 3 矮壮素 缩节胺在 4 种基质中的线性方程 相关系数 检出限和定量限 Linear equations, correlation coefficient, LOD and LOQ of chlormequat and mepiquat chloride in 4 matrixes. r LOD/(μg/L) LOQ/(μg/L) Y=1038.6X+224.7 0.9991 0.2 0.5 Y=1135.4X-92.2 0.9993 0.2 0.5 Y=836.2X+83.4 0.9990 0.4 1.2 Y=967.5X+122.5 0.9995 0.2 0.6 Y=1102.9X-44.7 0.9991 0.2 0.5 Y=1010.6X+111.7 0.9992 0.2 0.5 Y=1219.7X-62.5 0.9994 0.2 0.5 Y=1196.2X+89.7 0.9989 0.2 0.5
3688 5, -, ; -,, 3.2 色谱条件的优化 C 18, 20%,,, (40%),, A HCOOH NH 4 AC, ph, HCOOH 0.1% 0.2%,,,,, 0.2% HCOOH, 4 1 3.3 线性范围和检出限 2.2.1 0.22 μm, (Y), (X),,, (LOD) (LOQ) 4 3 3.4 精密度和加标回收率 4, 5, 1.3, ( 4) 3.5 实际样品检测 5 2, 5, 5,,,, 0.2 μg/kg, 0.4 μg/kg, 6#,,,, Table 4 表 4 矮壮素 缩节胺在 4 种样品中的加标回收率和精密度 (n=5) Recoveries and precision of chlormequat and mepiquat in 4 samples (n=5) /(mg/kg) /% RSD/% /(mg/kg) /% RSD/% 1.0 97.2 3.4 1.0 95.8 2.7 5.0 98.7 2.2 5.0 97.3 3.5 10.0 96.3 3.1 10.0 97.8 1.9 0.1 95.2 5.2 0.1 97.5 3.2 0.5 96.9 3.6 0.5 95.3 3.8 1.0 97.3 3.2 1.0 97.2 2.6 0.1 92.2 6.7 0.1 94.4 4.4 0.5 94.8 5.3 0.5 94.7 3.8 1.0 93.4 4.2 1.0 96.2 3.5 0.1 98.6 2.1 0.1 97.2 2.8 0.5 101.2 2.9 0.5 95.8 3.4 1.0 95.7 3.4 1.0 97.9 2.3
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