TY - JOUR

T1 - Liquid chromatography quadrupole-Orbitrap mass spectrometry for the simultaneous analysis of advanced glycation end products and protein-derived cross-links in food and biological matrices

AU - Poojary, Mahesha M.

AU - Zhang, Wei

AU - Greco, Ines

AU - De Gobba, Cristian

AU - Olsen, Karsten

AU - Lund, Marianne N.

PY - 2020

Y1 - 2020

N2 - Advanced glycation end products (AGEs) and protein cross-links have been extensively investigated in both food and biomedical fields over the past years. Although there are a few chromatographic and immunological methods for the analysis of selected AGEs, there is no method available for comprehensive simultaneous analysis of major AGEs found in processed foods and biological samples. In the present study, we have reported a validated UHPLC-MS/MS method for simultaneous identification and quantification of 15 different AGEs, furosine (an indicator of Amadori products), 2 protein-derived cross-links (lanthionine and lysinoalanine) and 2 amino acids (Lys and Arg). The analytes were separated on a reversed phase C-18 column and quantified accurately based on the isotope dilution method, where 9 stable isotope-labelled internal standards were used to quantify 20 different analytes using an Orbitrap mass analyzer. The method showed acceptable linearity, accuracy and precision. The LOD and LOQ values in plasma were in the range of 0.30–19.02 and 0.87–57.06 ng/mL, respectively. The recovery values at the three spiked levels were in the range of 71–110%, with some exceptions. The intraday and interday precision were in the range of 1.5–13.2%, however, quantification of N-ɛ-(carboxymethyl)lysine accompanied slightly higher interday precision (30.7%). The applicability of the method was successfully assessed by analyzing AGEs and protein cross-links in six different complex matrices including Ultra-High Temperature (UHT) processed milk, roasted chicken breast meat, roasted chicken skin, roasted pork liver, bovine plasma and perfusion liquid.

AB - Advanced glycation end products (AGEs) and protein cross-links have been extensively investigated in both food and biomedical fields over the past years. Although there are a few chromatographic and immunological methods for the analysis of selected AGEs, there is no method available for comprehensive simultaneous analysis of major AGEs found in processed foods and biological samples. In the present study, we have reported a validated UHPLC-MS/MS method for simultaneous identification and quantification of 15 different AGEs, furosine (an indicator of Amadori products), 2 protein-derived cross-links (lanthionine and lysinoalanine) and 2 amino acids (Lys and Arg). The analytes were separated on a reversed phase C-18 column and quantified accurately based on the isotope dilution method, where 9 stable isotope-labelled internal standards were used to quantify 20 different analytes using an Orbitrap mass analyzer. The method showed acceptable linearity, accuracy and precision. The LOD and LOQ values in plasma were in the range of 0.30–19.02 and 0.87–57.06 ng/mL, respectively. The recovery values at the three spiked levels were in the range of 71–110%, with some exceptions. The intraday and interday precision were in the range of 1.5–13.2%, however, quantification of N-ɛ-(carboxymethyl)lysine accompanied slightly higher interday precision (30.7%). The applicability of the method was successfully assessed by analyzing AGEs and protein cross-links in six different complex matrices including Ultra-High Temperature (UHT) processed milk, roasted chicken breast meat, roasted chicken skin, roasted pork liver, bovine plasma and perfusion liquid.

KW - AGEs

KW - Maillard reaction products

KW - Method validation

KW - Non-enzymatic glycosylation

KW - Parallel reaction monitoring

KW - Protein glycation

U2 - 10.1016/j.chroma.2019.460767

DO - 10.1016/j.chroma.2019.460767

M3 - Journal article

C2 - 31839352

AN - SCOPUS:85076513823

VL - 1615

JO - Journal of Chromatography

JF - Journal of Chromatography

SN - 0301-4770

M1 - 460767

ER -