TY - JOUR
T1 - Development and application of a stable isotope dilution analysis for the quantitation of advanced glycation end products of creatinine in biofluids of type 2 diabetic patients and healthy volunteers
AU - Kunert, Christof
AU - Skurk, Thomas
AU - Frank, Oliver
AU - Lang, Roman
AU - Hauner, Hans
AU - Hofmann, Thomas
PY - 2013/3/5
Y1 - 2013/3/5
N2 - N-(1-Methyl-4-oxoimidazolidin-2-ylidene) α-amino acids were recently identified in roasted meat as so far unknown advanced glycation end products (AGEs) of creatinine. For the first time, this paper reports on the preparation of 13C-labeled twin molecules of six N-(1-methyl-4-oxoimidazolidin-2- ylidene) α-amino acids and the development of a stable isotope dilution analysis (SIDA) for their simultaneous quantitation in meat, plasma, and urine samples by means of HPLC-MS/MS. Method validation demonstrated good precision (<14% RSD) and accuracy (97-118%) for all analytes and a lower limit of quantitation of 1 pg injected onto the column. The SIDA was applied to monitor plasma appearance and urinary excretion of these AGEs in type 2 diabetes mellitus patients (DM, n = 7) and healthy controls (n = 10) prior to and after ingestion of a bolus of processed beef meat. Interestingly, the basal concentration of N-(1-methyl-4-oxoimidazolidin-2-ylidene) aminopropionic acid was elevated in plasma and urine of DM patients compared to healthy individuals. Further, ingestion of processed meat led to a significantly higher concentration of this AGE in biofluids from DM patients when compared to healthy controls. These findings suggest a favored in vivo formation, as demonstrated by physiological model incubations of creatinine and carbohydrates (37 C, pH 7.4), or a more efficient dietary up-take of N-(1-methyl-4-oxoimidazolidin-2- ylidene) α-amino acids in hyperglycemic diabetes patients.
AB - N-(1-Methyl-4-oxoimidazolidin-2-ylidene) α-amino acids were recently identified in roasted meat as so far unknown advanced glycation end products (AGEs) of creatinine. For the first time, this paper reports on the preparation of 13C-labeled twin molecules of six N-(1-methyl-4-oxoimidazolidin-2- ylidene) α-amino acids and the development of a stable isotope dilution analysis (SIDA) for their simultaneous quantitation in meat, plasma, and urine samples by means of HPLC-MS/MS. Method validation demonstrated good precision (<14% RSD) and accuracy (97-118%) for all analytes and a lower limit of quantitation of 1 pg injected onto the column. The SIDA was applied to monitor plasma appearance and urinary excretion of these AGEs in type 2 diabetes mellitus patients (DM, n = 7) and healthy controls (n = 10) prior to and after ingestion of a bolus of processed beef meat. Interestingly, the basal concentration of N-(1-methyl-4-oxoimidazolidin-2-ylidene) aminopropionic acid was elevated in plasma and urine of DM patients compared to healthy individuals. Further, ingestion of processed meat led to a significantly higher concentration of this AGE in biofluids from DM patients when compared to healthy controls. These findings suggest a favored in vivo formation, as demonstrated by physiological model incubations of creatinine and carbohydrates (37 C, pH 7.4), or a more efficient dietary up-take of N-(1-methyl-4-oxoimidazolidin-2- ylidene) α-amino acids in hyperglycemic diabetes patients.
UR - http://www.scopus.com/inward/record.url?scp=84874608976&partnerID=8YFLogxK
U2 - 10.1021/ac303684v
DO - 10.1021/ac303684v
M3 - Article
C2 - 23379726
AN - SCOPUS:84874608976
SN - 0003-2700
VL - 85
SP - 2961
EP - 2969
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 5
ER -