Barefeet Analytics Pvt. Ltd, & MassTech LAS Applications Development Laboratory, 100 NCL Innovation Park, Dr. Homi Bhabha Rd., Pune 411 008, India.
National Reference Laboratory, ICAR-National Research Centre for Grapes, P.O. Manjri Farm, Pune 412 307, India.
J AOAC Int. 2022 Jun 29;105(4):1043-1050. doi: 10.1093/jaoacint/qsac012.
Aflatoxin M1 (AFM1) is a carcinogenic hydroxylated metabolite commonly found in milk. It is relatively stable toward decontamination procedures posing a major health risk, and it requires an international regulatory mandate of detection at trace levels.
To develop a high-throughput, reliable, and compliant method for the identification of AFM1 in milk samples using atmospheric pressure-matrix assisted laser desorption/ionization (AP-MALDI) selected reaction monitoring (SRM) quantitation.
The milk sample was diluted in water and cleaned with immunoaffinity chromatography (IAC), followed by analysis using AP-MALDI hyphenated with a triple quadrupole mass spectrometer for SRM.
A fast and reliable AP-MALDI SRM quantitative method was developed for the determination of AFM1 with analysis time of 1 min per sample. The diagnostic product ions of AFM1 at 273.1 u and 229.2 u were monitored during the SRM. The calibration curves yielded excellent linearity (R2 = 0.99) with good recoveries for quality control samples (97-106%). The ion ratios of the qualifier to quantifier displayed excellent RSD (1-7.8%) for n = 3.
The developed method provided rapid quantification for AFM1. The fast AP-MALDI SRM method can allow analysis of AFM1 in a large number of milk samples. Given the time required for analysis, cost-effectiveness, and superior analytical performance, this method can be adopted in commercial food testing laboratories.
Aflatoxins (AF) are a major health risk. Speedy analysis of large sample sizes from food is a risk mitigation strategy but remains an unmet need. Quantitative, chromatography-free, and internal standard-free AP-MALDI SRM based analysis of AF is a high-throughput and cost-efficient alternative. Satisfactory performance was achieved for quantitative AP-MALDI SRM analysis of AFM1 in milk subsequent to a simple sample clean-up step.
黄曲霉毒素 M1（AFM1）是一种常见于牛奶中的致癌羟化代谢物。它对去污程序具有相对稳定性，对痕量水平的检测需要国际监管授权。
开发一种高通量、可靠且符合法规的方法，用于使用大气压基质辅助激光解吸/电离（AP-MALDI）选择反应监测（SRM）定量法鉴定牛奶样品中的 AFM1。
将牛奶样品在水中稀释，并用免疫亲和色谱（IAC）进行净化，然后使用与三重四极杆质谱联用的 AP-MALDI 进行 SRM 分析。
开发了一种快速可靠的 AP-MALDI SRM 定量方法，用于测定 AFM1，每个样品的分析时间为 1 分钟。在 SRM 中监测 AFM1 的诊断产物离子 273.1 u 和 229.2 u。校准曲线具有极好的线性（R2=0.99），质控样品的回收率良好（97-106%）。n=3 时，质量标记物与定量标记物的离子比显示出极好的 RSD（1-7.8%）。
所开发的方法为 AFM1 提供了快速定量。快速的 AP-MALDI SRM 方法可以允许对大量牛奶样品进行分析。鉴于分析所需的时间、成本效益和卓越的分析性能，该方法可在商业食品检测实验室中采用。
黄曲霉毒素（AF）是一个主要的健康风险。快速分析大量食品样品是一种风险缓解策略，但仍然是一个未满足的需求。基于 AP-MALDI SRM 的无色谱、无内标定量分析是一种高通量且具有成本效益的替代方法。在经过简单的样品净化步骤后，成功实现了牛奶中 AFM1 的定量 AP-MALDI SRM 分析的令人满意的性能。
