Multiplex Quantification of Protein Toxins in Human Biofluids and Food Matrices Using Immunoextraction and High-Resolution Targeted Mass Spectrometry

详细信息    查看全文

• 作者：Mathieu Dupr茅 ; Benoit Gilquin ; Fran莽ois Fenaille ; C茅cile Feraudet-Tarisse ; Julie Dano ; Myriam Ferro ; St茅phanie Simon ; Christophe Junot ; Virginie Brun ; Fran莽ois Becher
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：August 18, 2015
• 年：2015
• 卷：87
• 期：16
• 页码：8473-8480
• 全文大小：353K
• ISSN：1520-6882

文摘

The development of rapid methods for unambiguous identification and precise quantification of protein toxins in various matrices is essential for public health surveillance. Nowadays, analytical strategies classically rely on sensitive immunological assays, but mass spectrometry constitutes an attractive complementary approach thanks to direct measurement and protein characterization ability. We developed here an innovative multiplex immuno-LC-MS/MS method for the simultaneous and specific quantification of the three potential biological warfare agents, ricin, staphylococcal enterotoxin B, and epsilon toxin, in complex human biofluids and food matrices. At least 7 peptides were targeted for each toxin (43 peptides in total) with a quadrupole-Orbitrap high-resolution instrument for exquisite detection specificity. Quantification was performed using stable isotope-labeled toxin standards spiked early in the sample. Lower limits of quantification were determined at or close to 1 ng路mL^鈥?. The whole process was successfully applied to the quantitative analysis of toxins in complex samples such as milk, human urine, and plasma. Finally, we report new data on toxin stability with no evidence of toxin degradation in milk in a 48 h time frame, allowing relevant quantitative toxin analysis for samples collected in this time range.