7 4 Vol. 7 No. 4 2016 4 Journal of Food Safety and Quality Apr., 2016 - - 16 杜娟 *, 马娇, 赵文霞, 宋杰 (, 100094) 摘要 : 目的 - -(GC-MS) 16 方法,,, GC-MS (SIM), 结果 500.0 1000.0 2500.0 μg/kg, 16 67.4%~110.5%, 0.3%~12.7% ; 16 60.5%~108.3%, 0.6%~12.7% 50.0~1000.0 μg/kg, 0.999, 500.0 μg/kg 结论, PAEs 关键词 : -; ; ; Determination of 16 kinds of phthalate esters in edible oils by gas chromatography-mass spectrometry with isotopic internal standards DU Juan *, MA Jiao, ZHAO Wen-Xia, SONG Jie (China National Food Quality & Safety Supervision and Inspection Center, Beijing 100094, China) ABSTRACT: Objective To establish a method for determination of 16 kinds of phthalate esters (PAEs) in edible oils by gas chromatography-mass spectrometry (GC-MS) with isotopic dilution method. Methods The samples were dissolved with n-hexane and extracted with acetonitrile, and then purified by solid-phase column. Using GC-MS with selected ion monitoring (SIM), quantitative analysis was performed based upon isotopic internal method. Results The spiked recoveries at levels of 500.0, 1000.0, 2500.0 μg/kg used soybean oil and butter samples as blank matrix were respectively in the range of 67.4%~110.5% and 60.5%~108.3%. The relative standard deviations (RSDs) were respectively in the range of 0.3%~12.7% and 0.6%~12.7%. The experimental results showed that 16 kinds of PAEs had good linear relations in the range of 50.0~1000.0 μg/kg, the correlation coefficients were better than 0.999, and the limits of detection (LODs) of 16 PAEs were 500.0 μg/kg. Conclusion The sensitivity, precision and LOD can meet the requirements for the determination of the practical samples. This method can be used for the simultaneous detection of PAEs in edible oil samples. KEY WORDS: gas chromatography-mass spectrometry; phthalate esters; edible oil; isotopic dilution method * 通讯作者 :,, E-mail: dujuanwenling@163.com *Corresponding author: DU Juan, Intermediate Engineer, China National Food Quality & Safety Supervision and Inspection Center, Beijing 100094, China. E-mail: dujuanwenling@163.com
1490 7 1 引言 (phthalate esters, PAEs),,,,,, PAEs [1],, [2], 20% ~ 50% [3], PAEs, PAEs,,, PAEs,, PAEs, PAEs, [4],, PAEs [5], PAEs,,,, PAEs,, PAEs,,, [6] PAEs (gas chromatography, GC) [7] - (gas chromatography-mass spectrometry, GC-MS) [8] - (gas chromatography with tandem mass spectrometry, GC-MS/MS) [9] (high performance liquid chromatography, HPLC) [10] (liquid chromatography-mass spectrometry, LC-MS) [11] - (high performance liquid chromatography with tandem mass spectrometry, HPLC-MS/MS) [12], GB/T 21911-2008 16 PAEs 1.5 mg/kg, [13], 0.5 mg/kg, ; (primary secondary amine, PSA),, PAEs [14] PAEs,,, - 16, 2 材料与方法 2.1 材料, 9, 10 2.2 试剂与仪器 16 (1000 μg /ml): (DMP)(DEP) (DIBP)(DBP) (2-) (DMEP) (4- -2 ) (BMPP) (2-) (DEEP) (DPP)(DHXP) (BBP) (2- ) (DBEP) (DCHP) (2- ) (DEHP)(DPHP) (DNOP)(DNP), 99%,, Accustandard 4 16 (100 μg/ml): D 4 - (D 4 -DMP) D 4 - (D 4 -DEP) D 4 -(D 4 -DIBP) D 4 - (D 4 -DBP) D 4 - (2-) (D 4 -DMEP) D 4 - (4- -2- ) (D 4 -BMPP) D 4 -(2-) (D 4 -DEEP) D 4 - (D 4 -DPP) D 4 - (D 4 -DHXP) D 4 - (D 4 -BBP) D 4 - (2-) (D 4 -DBEP) D 4 - (D 4 -DCHP) D 4 -(2- ) (D 4 -DEHP) D 4 - (D 4 -DPhP) D 4 - (D 4 -DNOP) D 4 - (D 4 -DNP), 95.0 %, Sigma 4 (, Honeywell ); 7890A-5975C / GC/MS( EI, Agilent ); BS224s ( Sartorius ); 80-2 ( 壇 ); Vortex-Genie2 ( Scientific Industries ) PSA (6 ml): 2.3 实验方法 2.3.1 标准溶液的配制 16 (10 μg/ml):
4, : - - 16 1491 1 ml 16 (1000 μg/ml) 100 ml, 10 μg/ml 16 D 4 (10 μg /ml): D 4-16 ( 0.1 mg) 10 μg /ml, 18, 16 D 4 (1 μg /ml): 10 μg/ml D 4-16 1.0 ml 10 ml,, 1.0 μg/ml, 4, 2.3.2 样品的前处理 (1) 0.5 g, 10 ml, 25 μl D 4 - (10 μg/ml), 1 ml, 1 min, 5 ml, 2 min, 20 min, 3000 r/min 3 min, 5 ml, 2, 40, 2 ml, (2) PSA (solid phase extraction, SPE) 5 ml 5 ml PSA SPE, ;, 5 ml 5 ml - (4:96, V:V), 40 0.5 ml,, 2 ml,, GC-MS (3) ( ):,, 2.3.3 仪器分析条件 (1) DB-5MS (30 m 0.25 mm, 0.25 μm, J&W ) : 60, 1 min, 20 /min 220, 1 min, 5 /min 280, 10 min, 20 /min 300, 4 min (2) (EI); 70 ev; () 260 ; 1 μl; ( >99.999%); 280 ; (SIM) 1 3 GC-MS, 16 PAEs SIM 1~2 16 0.1 mg/l PAEs ( 0.1 mg/l) 1 Table 1 表 1 16 种 PAEs 的保留时间和质谱碎片离子 Retention time and mass fragment ions of 16 kinds of PAEs (min) (m/z) (m/z) 1 (DMP) 7.604 163, 77, 194, 133 163 2 (DEP) 8.460 149, 177, 105, 193 149 3 (DIBP) 10.127 149, 223, 104, 167 149 4 (DBP) 10.835 149, 223, 205, 104 149 5 (2-) (DMEP) 11.162 59, 149, 104, 76 149 6 (4- -2- ) (BMPP) 11.868 149, 167, 85, 251 149 7 (2-) (DEEP) 12.181 45, 72, 149, 104, 193 149 8 (DPP) 12.534 149, 237, 219, 104 149 9 (DHXP) 14.601 149, 251, 104, 233 149 10 (BBP) 14.751 149, 91, 206, 132 149 11 (2-) (DBEP) 16.163 149, 101, 85, 193 149 12 (DCHP) 16.797 149, 167, 249, 104 149 13 (2- ) (DEHP) 17.034 149, 167, 279, 113 149 14 (DPHP) 17.174 225, 77, 104, 153 225 15 (DNOP) 19.392 149, 279, 104, 261 149 16 (DNP) 21.878 149, 293, 167, 275 149
1492 7 Table 2 表 2 16 种 PAEs 同位素内标保留时间和质谱碎片离子 Retention time and mass fragment ions of internal standards of 16 kinds of PAEs (min) (m/z) (m/z) 1 (D 4 -DMP) 7.604 167, 77, 198, 137 167 2 (D 4 -DEP) 8.460 153, 181, 109, 197 153 3 (D 4 -DIBP) 10.127 153, 227, 108, 171 153 4 (D 4 -DBP) 10.835 153, 227, 209, 108 153 5 (2-) (D 4 -DMEP) 11.162 59, 153, 108, 76 153 6 (4- -2- ) (D 4 -BMPP) 11.868 153, 171, 85, 255 153 7 (2-) (D 4 -DEEP) 12.181 45, 72, 153, 108, 197 153 8 (D 4 -DPP) 12.534 153, 241, 223, 108 153 9 (D 4 -DHXP) 14.601 153, 255, 108, 237 153 10 (D 4 -BBP) 14.751 153, 91, 210, 136 153 11 (2-) (D 4 -DBEP) 16.163 153, 105, 85, 197 153 12 (D 4 -DCHP) 16.797 153, 171, 253, 108 153 13 (2- ) (D 4 -DEHP) 17.034 153, 171, 283, 117 153 14 (D 4 -DPHP) 19.840 229, 77, 108, 157 229 15 (D 4 -DNOP) 19.392 153, 283, 108, 265 153 16 (D 4 -DNP) 21.878 153, 297, 171, 279 153 1 16 1000.0 μg/l PAEs ( 125.0 μg/l) Fig. 1 Total ion chromatogram of 16 kinds of PAEs with the concentration of 1000.0 μg/l(isotopic internal standards, 125.0 μg/l) : 1.DMP(D 4 -DMP); 2.DEP(D 4 -DEP); 3.DIBP(D 4 -DIBP); 4.DBP(D 4 -DBP); 5.DMEP(D 4 -DMEP); 6.BMPP(D 4 -BMPP); 7.DEEP(D 4 -DEEP); 8.DPP(D 4 -DPP); 9.DHXP(D 4 -DHXP); 10.BBP(D 4 -BBP); 11.DBEP(D 4 -DBEP); 12.DCHP(D 4 -DCHP); 13.DEHP(D 4 -DEHP); 14.DPhP; 15.DNOP(D 4 -DNOP); 16. D 4 -DPhP; 17.DNP(D 4 -DNP).
4, : - - 16 1493 3 结果与讨论 3.1 提取溶剂的选择 PAEs,, PAEs,, ; ( 500.0 μg/kg),, 70%~110%,,,,,,,, 2 ml, 3.2 固相萃取 (SPE) 净化,,,, (gel permeation chromatography, GPC),,,, SPE,, [15], PSA,, 3.3 色谱条件的选择 PAEs, DNOP DNP,, DB-5MS 16 PAEs,, GC-MS,,, 3.3 ( 1) 3.4 方法学参数验证 3.4.1 线性范围及方法检测限, 16 (10 μg/ml)5.0 10.0 25.0 50.0 100.0 μl 16 D 4 (1 μg/ml)125.0 μl, 1 ml, 16 50.0 100.0 250.0 500.0 1000.0 μg/l, 125.0 μg/l, GC-MS, ( 3) PAEs 50.0~1000.0 μg/l, r 0.999 (S/N) 3, 500.0 μg/kg,, 16 PAEs (S/N) 3, 500.0 μg/kg Table 3 表 3 16 种 PAEs 的线性方程 相关系数及方法检测限 Linear equations, correlation coefficient (r) and limit of detection (LOQ) of 16 kinds of PAEs r LOD(μg/kg) 1 DMP Y=1.177X+1.622 10 3 0.9999 500.0 2 DEP Y=1.083X+1.092 10 2 0.9999 500.0 3 DIBP Y=1.081X+7.953 10 2 0.9999 500.0 4 DBP Y=1.062X+8.992 10 2 0.9999 500.0 5 DMEP Y=0.227X+1.185 10 1 0.9999 500.0 6 BMPP Y=2.704X 3.689 10 3 0.9999 500.0 7 DEEP Y=1.603X+1.434 10 0 0.9999 500.0 8 DPP Y=0.986X+9.709 10 4 0.9999 500.0 9 DHXP Y=1.007X 1.757 10 2 0.9999 500.0 10 BBP Y=1.059X+1.108 10 2 0.9999 500.0 11 DBEP Y=1.047X 1.073 10 2 0.9999 500.0 12 DCHP Y=0.944X 4.237 1 0 3 0.9999 500.0 13 DEHP Y=0.996X+1.619 10 2 0.9999 500.0 14 DPHP Y=2.301X 2.176 10 1 0.9999 500.0 15 DNOP Y=1.026X 1.862 10 2 0.9999 500.0 16 DNP Y=1.111X 1.476 10 2 0.9999 500.0
1494 7 3.4.2 方法的精密度和回收率, PAEs 3 (500.0 1000.0 2500.0 μg/kg), 6, 16 D 4 (1 μg/ml)125.0 μl, 125.0 μg/kg, (RSD),,, 3 67.4%~110.5%, RSD 12.7%, 3 60.5%~108.3%, RSD 12.7%, ( 4) 3.4.3 样品分析 2015 1,, 90, PAEs,, PAEs, DBP DEHP, 14 PAEs, 5 DBP 1.74 mg/kg, DEHP 1.26mg/kg,, PE PP PAEs, ;, Table 4 表 4 大豆油和黄油 3 水平加标回收实验结果 (n=6) Recovery and relative standard deviations (RSDs) at 3 spiked level in soybean oil and butter (n=6) /(μg/kg) /% RSD/% /% RSD/% 500.0 110.1 3.7 108.3 1.4 DMP 1250.0 106.8 2.3 106.2 2.1 2500.0 97.9 3.8 105.3 2.5 500.0 106.6 3.5 104.1 1.0 DEP 1250.0 104.1 2.6 104.3 1.7 2500.0 96.9 4.7 97.6 2.2 500.0 103.3 3.2 99.7 1.7 DAP 1250.0 100.9 3.5 97.6 3.6 2500.0 96.3 3.7 91.3 3.4 500.0 96.5 3.0 101.0 6.7 DIBP 1250.0 101.6 0.3 104.4 2.6 2500.0 94.5 4.8 95.4 3.6 500.0 79.4 4.7 92.2 6.7 DBP 1250.0 93.3 1.4 97.8 4.9 2500.0 92.7 3.8 95.0 4.6 500.0 94.1 6.8 96.6 3.3 DMEP 1250.0 96.9 1.0 94.8 3.0 2500.0 89.4 7.1 106.7 2.4 500.0 105.6 5.3 79.1 4.2 BMPP 1250.0 92.1 2.5 79.7 5.0 2500.0 94.3 6.9 73.8 8.8 500.0 103.9 2.1 91.3 2.2 DEEP 1250.0 101.9 1.5 88.9 0.8 2500.0 94.2 4.2 86.9 2.1
4, : - - 16 1495 /(μg/kg) 续表 4 /% RSD/% /% RSD/% 500.0 106.3 3.6 100.7 0.6 DPP 1250.0 103.9 2.7 103.0 1.7 2500.0 96.4 4.7 93.6 1.6 500.0 105.0 3.7 99.6 0.9 DHXP 1250.0 102.6 2.5 102.1 1.5 2500.0 96.4 5.2 93.0 1.4 500.0 103.3 3.4 94.8 2.7 BBP 1250.0 103.2 3.1 97.0 2.2 2500.0 96.0 5.6 90.4 3.5 500.0 110.1 4.0 100.8 2.8 DBEP 1250.0 104.8 3.3 101.9 3.8 2500.0 96.6 6.0 91.8 2.0 500.0 104.7 3.9 99.3 1.0 DCHP 1250.0 102.7 3.0 104.3 2.3 2500.0 95.8 5.2 92.8 2.1 500.0 83.3 12.7 99.5 9.1 DEHP 1250.0 99.9 7.1 93.4 11.7 2500.0 97.2 12.7 86.8 3.5 500.0 80.3 6.8 69.4 9.9 DPHP 1250.0 79.4 1.1 76.6 12.7 2500.0 67.4 5.7 61.7 4.7 500.0 105.2 3.6 98.8 2.4 DNOP 1250.0 101.5 2.5 99.1 3.8 2500.0 96.3 5.0 93.4 3.4 500.0 110.5 5.0 60.5 7.2 DNP 1250.0 105.6 2.4 86.5 9.8 2500.0 98.8 5.9 63.0 11.8 Table 5 表 5 90 份油脂样品 16 种 PAEs 的检出情况 Results obtained for determination of 16 kinds of PAEs in 90 oil samples DBP/mg/kg ND ND ND~1.57 ND~1.74 ND~1.09 ND ND~1.37 ND ND /% 0 0 0.1 0.1 0.1 0 0.1 0 0 DEHP/mg/kg ND~1.22 ND~1.26 ND ND ND ND ND~1.06 ND~1.11 ND /% 0.1 0.1 0 0 0 0 0.1 0.1 0 : ND
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