Eight marker peptides in meat were identified and quantitatively determined by ultra-high pressure liquid chromatography coupled with quadrupole time-of-flight mass spectrometry,and the effects of chromatographic separation conditions and mass spectrometry parameters on the determination of marker peptides in meat were investigated. The samples were separated by a C18 column, and gradient elution was performed with 0.1% formic acid in water and acetonitrile (containing 0.1% formic acid) as mobile phase. Then, the electrospray positive ion mode (ESI+) was used to detect the marker peptides, and quantitative analysis and data collection were performed by pseudo-multiple reaction monitoring by time-of-flight mass spectrometry (TOF-MRM). Results showed that the parent ions of the targeted marker peptides had a 2+ or 3+ charge, which showed high signal intensity and good stability. And the fragment ions of each marker peptide in the mass spectrometry was mainly y ion. Moreover, the fragment ions with 3+ charge were accompanied by the formation of 2+ charges. The TOF-MRM acquisition mode and target enhancement increased the detection sensitivity of target ions. Under the optimized conditions, the separation of 8 marker peptides was good and the mass signals were strong. There was a good linear relationship between the concentration of 8 marker peptides standards and detection response, and the correlation coefficients were greater than 0.98. The limit of detection (LOD) for analysts was in the range of 0.19 - 1.50 μg/L, and the optimized TOF-MRM method was successfully applied to the analysis of marker peptides in meat samples due to the high accuracy, precision and repeatability. The application of marker peptides on 7 class beef meatball products showed that five marker peptides could be detected including 2 beef, 1 pork and 2 chicken meat, which indicated that cheaper chicken or pork was spiked in beef meatball samples. The abundance of marker peptides had no correlation with the price. However, no duck-specific marker peptides were found. The established species-specific peptide detection method provided powerful technical support for the identification of protein species origin.
WANG Zhonghe
,
LI Xiaoting
,
HU Wenmei
,
WANG Jun
. Determination of peptide markers of meat species by ultra-high pressure liquid chromatography coupled with high resolution mass spectrometry[J]. Food and Fermentation Industries, 2021
, 47(18)
: 258
-266
.
DOI: 10.13995/j.cnki.11-1802/ts.028030
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