Quantification of Cry1Ab in genetically modified maize leaves by liquid chromatography multiple reaction monitoring tandem mass spectrometry using 18O stable isotope dilution.

Abstract	Cry1Ab is one of the most common Bacillus thuringiensis (Bt) proteins in genetically modified crops, which exhibits strong resistance against insect pests. In the present study, a sensitive and precise liquid chromatography stable isotope dilution multiple reaction monitoring tandem mass spectrometry (LC-SID-MRM-MS) assay was developed and validated to quantify the amount of Cry1Ab expression in transgenic maize leaves. The measurement of protein was converted to measurement of unique peptides to Cry1Ab protein. Two peptides unique to Cry1Ab were synthesized and labeled in H(2)(18)O to generate (18)O stable isotope peptides as internal standards. The validated method obtained superior specificity and good linearity. And the inter- and intra-day precision and accuracy for all samples were satisfactory. The results demonstrated Cry1Ab protein was 31.7 ± 4.1 μg g(-1) dry weight in Bt-176 transgenic maize leaves. It proved that the novel LC-SID-MRM-MS method was sensitive and selective to quantify Cry1Ab in the crude extract without time-consuming pre-separation or purification procedures.
Authors	Yongqian Zhang, Chengjun Lai, Rui Su, Mei Zhang, Yan Xiong, Hong Qing, Yulin Deng
Journal	The Analyst (Analyst) Vol. 137 Issue 11 Pg. 2699-705 (Jun 07 2012) ISSN: 1364-5528 [Electronic] England
PMID	22543512 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	Bacillus thuringiensis Toxins Bacterial Proteins Endotoxins Hemolysin Proteins Oxygen Isotopes Peptides Recombinant Proteins insecticidal crystal protein, Bacillus Thuringiensis
Topics	Amino Acid Sequence Bacillus thuringiensis Toxins Bacterial Proteins (analysis, genetics, metabolism) Chromatography, High Pressure Liquid Endotoxins (analysis, genetics, metabolism) Hemolysin Proteins (analysis, genetics, metabolism) Oxygen Isotopes (chemistry) Peptides (analysis) Plant Leaves (metabolism) Plants, Genetically Modified (metabolism) Recombinant Proteins (analysis, genetics, metabolism) Tandem Mass Spectrometry Zea mays (metabolism)

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