Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for the Simultaneous Quantification of N^τ-Methylhistidine, N-Acetyl-N^τ-Methylhistidine, and Creatinine in Urine.

IF 3.8 3区医学 Q2 NUTRITION & DIETETICS

Journal of Nutrition Pub Date : 2025-09-23 DOI:10.1016/j.tjnut.2025.09.024

Daichi Ijiri, Yoshio Nishimura, Ayumi Katafuchi, Saki Shimamoto, Miyu Kamimura, Ichiro Oshima, Namiko Nakamura, Shozo Tomonaga, Shinya Ishinhara, Akira Ohtsuka

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引用次数: 0

Abstract

Background: N^τ-Methylhistidine released endogenously from muscle into urine is an index of muscle protein breakdown, and part of N^τ-methylhistidine it is excreted as the acetylated derivative, N-acetyl-N^τ-methylhistidine).

Objective: To measure the urinary concentrations of N^τ-methylhistidine and N-acetyl-N^τ-methylhistidine, a quantitative method was established using liquid chromatography-tandem mass spectrometry (LC-MS/MS) operated in Q1 single-ion monitoring (SIM) mode and multiple reaction monitoring (MRM) mode with stable-isotope dilution analysis.

Methods: We synthesized N-acetyl-N^τ-methylhistidine and its isotopic analog for stable-isotope dilution analysis. Then, we validated a method for quantifying urinary concentrations of intact N^τ-methylhistidine, N-acetyl-N^τ-methylhistidine, and creatinine to precisely determine their concentrations in rats, humans, cattle, and dogs. Creatinine correction was applied to normalize for urine volume.

Results: For both SIM and MRM analysis, the acceptable linear ranges of detection were 5 pmol/mL to 10 nmol/mL with r² = 1.000 in SIM mode and MRM mode. The LC-MS/MS methods operated in both SIM and MRM transitions detected changes in the urinary excretion levels of N^τ-methylhistidine and N-acetyl-N^τ-methylhistidine in response to starvation and re-feeding of rats. The methods also detected urinary concentrations of N^τ-methylhistidine and N-acetyl-N^τ-methylhistidine in cattle, humans, and dogs.

Conclusions: These results suggest that the LC-MS/MS method operated in SIM mode and MRM mode can be used to measure urinary concentrations of N^τ-methylhistidine and N-acetyl-N^τ-methylhistidine.

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液相色谱-串联质谱法同时测定尿中n - τ-甲基组氨酸、n -乙酰- n - τ-甲基组氨酸和肌酐的建立和验证。

背景：n τ-甲基组氨酸从肌肉内源性释放到尿液中是肌肉蛋白分解的指标，部分n τ-甲基组氨酸作为乙酰化衍生物（n -乙酰化- n τ-甲基组氨酸）排出体外。目的：建立液相色谱-串联质谱（LC-MS/MS）联用Q1单离子监测（SIM）模式和多反应监测（MRM）模式，结合稳定同位素稀释分析，测定尿中n - τ-甲基组氨酸和n -乙酰- n -甲基组氨酸浓度的定量方法。方法：合成n -乙酰基- n τ-甲基组氨酸及其同位素类似物进行稳定同位素稀释分析。然后，我们验证了一种定量尿中完整n -甲基组氨酸、n -乙酰- n -甲基组氨酸和肌酐浓度的方法，以精确测定它们在大鼠、人、牛和狗中的浓度。应用肌酐校正使尿量正常化。结果：对于SIM和MRM分析，在SIM模式和MRM模式下，可接受的线性检测范围为5 pmol/mL至10 nmol/mL， r2 = 1.000。LC-MS/MS方法在SIM和MRM转换中检测了大鼠在饥饿和再喂养时尿中n -甲基组氨酸和n -乙酰- n -甲基组氨酸水平的变化。该方法还检测了牛、人类和狗尿液中n - τ-甲基组氨酸和n -乙酰- n -甲基组氨酸的浓度。结论：LC-MS/MS方法在SIM模式和MRM模式下可用于测定尿中n - τ-甲基组氨酸和n -乙酰- n -甲基组氨酸的浓度。

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来源期刊

Journal of Nutrition 医学-营养学

CiteScore

7.60

自引率

4.80%

发文量

260

审稿时长

39 days

期刊介绍： The Journal of Nutrition (JN/J Nutr) publishes peer-reviewed original research papers covering all aspects of experimental nutrition in humans and other animal species; special articles such as reviews and biographies of prominent nutrition scientists; and issues, opinions, and commentaries on controversial issues in nutrition. Supplements are frequently published to provide extended discussion of topics of special interest.