Laboratory of Toxicology, Faculty of Health Sciences, University of Brasília, 70910-900 Brasília, DF, Brazil.
J Chromatogr A. 2013 Aug 23;1304:61-8. doi: 10.1016/j.chroma.2013.06.049. Epub 2013 Jun 27.
The aims of this study were to optimize and validate a methodology for the simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 (AFB1, AFB2, AFG1, AFG2, AFM1) and ochratoxin A (OTA) in breast milk, and to analyze these mycotoxins in samples obtained from human milk banks in the Federal District, Brazil. The optimized analytical method was based on liquid-liquid extraction with low temperature purification (3.25mL of acidified acetonitrile+0.75mL of ethyl acetate), followed by analysis by high-performance liquid chromatography with fluorescence detector (HPLC/FLD) and a photochemical post-column reactor. Limits of quantification (LOQ) ranged from 0.005 to 0.03ng/mL, recoveries from 73 to 99.5%, and relative standard deviations (RSD) from 1.8 to 17.3%. The LLE-LTP extraction method was shown to be simple and cost-effective, since no columns were needed for clean-up. Only 2 of the 224 breast milk samples analyzed were positive for the mycotoxins, both samples containing AFB2 at the LOQ level (0.005ng/mL). The identity of the mycotoxin detected was confirmed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This result indicates that infants who are fed with breast milk from the milk banks are not at risk from aflatoxin and ochratoxin exposure.
本研究旨在优化并验证一种同时分析母乳中黄曲霉毒素 B1、B2、G1、G2、M1（AFB1、AFB2、AFG1、AFG2、AFM1）和赭曲霉毒素 A（OTA）的方法，并分析来自巴西联邦区母乳库的这些真菌毒素。优化后的分析方法基于低温净化的液液萃取（3.25mL 酸化乙腈+0.75mL 乙酸乙酯），然后通过高效液相色谱荧光检测器（HPLC/FLD）和光化学反应柱后反应器进行分析。定量限（LOQ）范围为 0.005 至 0.03ng/mL，回收率为 73%至 99.5%，相对标准偏差（RSD）为 1.8%至 17.3%。LLE-LTP 萃取方法简单且具有成本效益，因为不需要柱子进行净化。在所分析的 224 份母乳样本中，仅有 2 份样本的真菌毒素呈阳性，均为 AFB2 含量达到 LOQ 水平（0.005ng/mL）。通过液相色谱-串联质谱法（LC-MS/MS）确认了所检测到的真菌毒素的身份。这一结果表明，从母乳库中喂养婴儿的婴儿不会面临黄曲霉毒素和赭曲霉毒素暴露的风险。
