超高效液相色谱-串联质谱法测定牛奶中11种对苯二胺类抗氧化剂及其氧化产物
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作者单位:

1.南京医科大学 公共卫生学院,江苏 南京 211166;2.国家食品安全风险评估中心,北京 100021

作者简介:

刘光臣 男 在读研究生 研究方向为食品安全 E-mail: 13754908551@163.com

通讯作者:

李敬光 男 研究员 研究方向为食品安全 E-mail: lijg@cfsa.net.cn

中图分类号:

R155

基金项目:

国家重点研发计划(2023YFF1104802)


Determination of 11 p-phenylenediamine antioxidants and their oxidation products in milk by ultra performance liquid chromatography-tandem mass spectrometry
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Affiliation:

1.School of Public Health, Nanjing Medical University, Jiangsu Nanjing 211166, China;2.China National Center for Food Safety Risk Assessment, Beijing 100021, China

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    摘要:

    目的 建立固相萃取-超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定牛奶中11种对苯二胺类抗氧化剂(PPDs)及其氧化产物(PPD-Qs)的方法。方法 称取2.00 g牛奶样品,冷冻干燥后加入10 mL乙腈,振荡提取,低温离心。上清液使用Oasis HLB固相萃取柱除杂净化,合并上样液与净化液经氮气吹干复溶后,进行进样分析。采用含0.1%甲酸的水溶液和乙腈作为流动相进行梯度洗脱,WATERS ACQUITY BEH C18(2.1 mm×100 mm,1.7 μm)作为色谱分析柱,在正离子电喷雾(ESI+)多反应监测模式下检测,内标法定量。结果 在0.1~50 μg/L范围内,11种PPD及其氧化产物的相关系数(R2)均>0.990,方法检出限为0.004~0.152 μg/kg,定量限为0.013~0.507 μg/kg,3种不同加标浓度的回收率为84.4%~121.9%,日间和日内精密度良好(RSD<14%)。应用该方法检测不同国家和地区的28份牛奶样品,其中6PPD的检出率最高,阳性样品浓度为0.05~2.27 μg/kg;DTPD的检出率为39%,阳性样品浓度为0.07~0.79 μg/kg;仅有两例样品中6PPD-Q的浓度高于定量限。结论 该方法具有良好的灵敏度、准确度和精密度,适用于牛奶样品中对苯二胺类抗氧化剂及其氧化产物的检测。

    Abstract:

    Objective To establish the first method for the simultaneous determination of 11 p-phenylenediamine antioxidants (PPDs) and their oxidation products in milk by solid-phase extraction (SPE) coupled with ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS).Methods A 2.00 g milk sample was weighed, freeze-dried, and extracted with 10 mL of acetonitrile. The mixture was shaken, then centrifuged at low temperature. The supernatant was purified using an Oasis HLB solid-phase extraction column. The combined sample and purified liquid were evaporated under nitrogen and resuspended for injection and analysis. Chromatographic separation was performed using a gradient elution with a mobile phase of 0.1% formic acid aqueous solution and acetonitrile. The analysis was carried out on a WATERS ACQUITY BEH C18 column (2.1 mm×100 mm, 1.7 μm). Detection was conducted in positive ion electrospray ionization (ESI+) multiple reaction monitoring (MRM) mode, and quantification was achieved using the internal standard method.Results The correlation coefficients (R2) for all 11 PPDs and their oxidation products were greater than 0.990 within the concentration range of 0.1-50 μg/L. The method detection limits ranged from 0.004 to 0.152 μg/kg, and the quantification limits ranged from 0.013 to 0.507 μg/kg. Recovery rates for three different spiked concentrations ranged from 84.4% to 121.9%, with good intra-day and inter-day precision (RSD<14%). When applied to 28 milk samples from different countries and regions, the detection rate for 6PPD was the highest, with positive concentrations ranging from 0.05 to 2.27 μg/kg. The detection rate for DTPD was 39%, with positive concentrations ranging from 0.07 to 0.79 μg/kg, and only two samples had 6PPD-Q concentrations above the limit of quantification.Conclusion This method demonstrates excellent sensitivity, accuracy, and precision, making it suitable for detecting p-phenylenediamine-based antioxidants and their oxidation products in milk samples.

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刘光臣,王清格,张磊,李敬光.超高效液相色谱-串联质谱法测定牛奶中11种对苯二胺类抗氧化剂及其氧化产物[J].中国食品卫生杂志,2025,37(8):715-721.

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  • 收稿日期:2025-07-07
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  • 在线发布日期: 2026-01-14
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