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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

机译:2型糖尿病患者和健康志愿者生物流体中肌酐高级糖基化终末定量的稳定同位素稀释分析方法的开发和应用

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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 ~(13)C-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℃, 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.
机译:N-(1-甲基-4-氧代咪唑啉丁-2-亚基)α-氨基酸最近在烤肉中被鉴定为迄今未知的肌酸酐高级糖化终产物(AGEs)。本文首次报道了〜(13)C标记的6个N-(1-甲基-4-氧代咪唑啉丁-2-亚基)α-氨基酸双分子的制备以及稳定同位素稀释剂的开发HPLC-MS / MS同时分析肉,血浆和尿液样品中的痕量氨基酸(SIDA)。方法验证表明,所有分析物均具有良好的精密度(<14%RSD)和准确度(97-118%),并且下限为进样到色谱柱的1 pg定量限。 SIDA用于监测大剂量加工牛肉的摄入前后的2型糖尿病患者(DM,n = 7)和健康对照组(n = 10)的血浆和这些AGEs的尿液排泄情况。有趣的是,与健康个体相比,DM患者的血浆和尿液中N-(1-甲基-4-氧代咪唑啉丁-2-亚基)氨基丙酸的基础浓度升高。此外,与健康对照组相比,加工肉类的摄入导致来自DM患者的生物流体中这种AGE的浓度显着升高。这些发现表明,在体内形成肌酸和碳水化合物(37℃,pH 7.4),或者更有效地从饮食中摄取N-(1-甲基-4-氧代咪唑啉丁-2-亚基)可以证明体内的形成。 )高血糖糖尿病患者的α-氨基酸。

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