Determination of 4 Polycyclic Aromatic Hydrocarbons in 3 Edible Oils by HPLC with Fluorescence Detection After Sample Pretreatment by Specific Enzymolysis
摘 要
称取食用油(动物油、植物油或经用于煎炸的食用油)2.0 g与pH 8.0磷酸盐缓冲溶液5 mL和脂肪酶0.2 g混匀后,于(37±2)℃振荡酶解2 h。加入碳酸钾1.0 g皂化样品中的脂肪,以及乙醇5 mL使在后续的提取中降低溶液的乳化现象。于此溶液中加入水5 mL及正己烷提取4种多环芳烃{PAHs,即苯并[a]蒽(BaA)、苯并[b]荧蒽(BbF)、苯并[k]荧蒽(BkF)和苯并[a]芘(BaP)} 2次,每次用正己烷15 mL,振荡5 min,离心5 min,移取上层提取液,合并后于40℃旋蒸至干。用乙腈溶解残渣并定容至1.0 mL,此溶液经0.22 μm滤膜过滤,取滤液供在仪器工作条件下进行高效液相色谱(HPLC)分离后测定。选择Dikma Plus C18色谱柱为固定相,以不同体积比的(A)水和(B)乙腈的混合液为流动相进行梯度洗脱。上述4种PAHs的保留时间依次为9.3,12.0,12.6,13.9 min。用荧光检测器分别测定其发射荧光的强度。在此4种PAHs中,BaA的荧光发射波长(λem)为380 nm,BbF、BkF和BaP的λem在406 nm波长处。分别取上述3种食用油的空白样品作为基体,加入加4种PAH的混合标准溶液系列制作标准曲线,测得其线性范围在相同的区间(0.1~5.0 μg·L-1),其检出限(3S/N)为0.03 μg·kg-1。分别以3种食用油样品作为基体,按标准加入法进行回收试验,测得其回收率均在90.0%以上,测定值的相对标准偏差(n=5)均小于9.0%。
Abstract
A portion (2.0 g) of edible oil sample (animal oil, vegetable oil or oil after use in frying) was mixed with 5 mL of pH 8.0 phosphate buffer solution and 0.2 g of lipase, and enzymolyzed for 2 h at (37±2)℃ under stirring. 1.0 g of K2CO3 and 5 mL of ethanol were added and mixed thoroughly to saponify the fat and to reduce emulsification of the solution during the following extraction. Four PAHs (BaA, BbF, BkF and BaP) in the solution after adding 5 mL of water were then extracted twice with n-hexane (15 mL for each extraction) by shaking for 5 min, and after centrifugation for 5 min, and the extracts of the upper phase were collected, combined and evaporated to dryness at 40℃. The residue was dissolved and diluted to 1.0 mL with acetonitrile. The solution was filtered through 0.22 μm filtering membrane and the filtrate was used for HPLC analysis under the intrumental working condition. Dikma Plus C18 chromatographic column was used as the stationary phase, and mixtures of (A) water and (B) acetonitrile in various ratios were used as mobile phases in the gradient elution. Retentiom times of 9.3, 12.0, 12.6, 13.9 min were found for the 4 PHAs respectively. Fluorescence detection (FLD) was made in the HPLC determination, and values of intensity of the emission fluorescence of the 4 analytes were measured at the λem of 380 nm for BaA and at the λem of 406 nm for BbF, BkF and BaP. Standard curves for the 4 analytes were prepared with the mixed standard solution of the 4 PHAs added separately to the 3 oil samples as matrixes, giving same linearity range between 0.1 and 5.0 μg·L-1. Values of detection limits (3S/N) found were same of 0.03 μg·kg-1 for the 4 PAHs. Tests for recovery were made by standard addition method using the 3 oil samples as matrixes, and values of recovery found were all over 90.0%, and values of RSDs (n=5) found were all below 9.0%.
中图分类号 O657.7 DOI 10.11973/lhjy-hx202006016
所属栏目 工作简报
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收稿日期 2019/10/7
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备注章再婷,工程师,研究方向为食品污染物分析
引用该论文: ZHANG Zaiting,YANGYan,WANG Yongjian,WANG Chunfang,LI Shuang,CHEN Shubing,CAO Guozhou. Determination of 4 Polycyclic Aromatic Hydrocarbons in 3 Edible Oils by HPLC with Fluorescence Detection After Sample Pretreatment by Specific Enzymolysis[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2020, 56(6): 713~718
章再婷,杨燕,王永健,王春芳,李双,陈树兵,曹国洲. 专一性酶解-高效液相色谱-荧光检测法测定3种食用油中4种多环芳烃[J]. 理化检验-化学分册, 2020, 56(6): 713~718
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参考文献
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【12】王建华,郭翠,庞国芳,等.GPC净化-同位素稀释内标定量GC-MS对植物油中多环芳烃的测定[J].分析测试学报, 2009,28(3):267-271.
【2】TAGHVAEE Z, PIRAVIVANAK Z, REZAEI K, et al. Determination of polycyclic aromatic hydrocarbons (PAHs) in olive and refined pomace olive oils with modified low temperature and ultrasound-assisted liquid-liquid extraction method followed by the HPLC/FLD[J]. Food Analytical Methods, 2016,9(5):1220-1227.
【3】CHEN Y C, CHEN B H. Determination of polycyclic aromatic hydrocarbons in fumes from fried chicken legs[J]. Journal of Agricultural and Food Chemistry, 2003,51(14):4162-4167.
【4】田玉霞,孟橘.食用油中多环芳烃的研究进展[J].中国油脂, 2012,37(3):69-73.
【5】DA SILVA S A, SAMPAIO G R, DA SILVA TORRES E A F. Optimization and validation of a method using UHPLC-fluorescence for the analysis of polycyclic aromatic hydrocarbons in cold-pressed vegetable oils[J]. Food Chemistry, 2017,221:809-814.
【6】赵佳莹,李晓敏,卢晓华,等.高效液相色谱-荧光检测法测定橄榄油中4种多环芳烃[J].色谱, 2017,35(7):748-754.
【7】黄华,吴颖,赵雅松,等.SPE净化-气相色谱-三重四极杆质谱对油脂中多环芳烃的测定[J].食品工业科技, 2014,35(7):308-311.
【8】吴春英,白鹭,谷风,等.UPLC-MS/MS测定地沟油中黄曲霉毒素和苯并芘[J].食品工业, 2017,38(1):285-288.
【9】朱捷,马桂娟,张瑶,等.QuEChERS/气相色谱三重四极杆质谱法测定枸杞籽油中多环芳烃类物质[J].中国油脂, 2019,44(6):89-94.
【10】王磊,樊蕊.凝胶色谱-气质联用测定花生油中苯并芘残留[J].食品工业, 2019,40(5):322-324.
【11】王溪,阮丽萍,李放,等.凝胶渗透色谱净化-高效液相色谱荧光法检测生肉中15种多环芳烃[J].江苏预防医学, 2019,30(3):259-261.
【12】王建华,郭翠,庞国芳,等.GPC净化-同位素稀释内标定量GC-MS对植物油中多环芳烃的测定[J].分析测试学报, 2009,28(3):267-271.
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