DOI: 10.1002/bmc.5724 ISSN:

A simplified D3‐creatine dilution method for skeletal muscle mass determination with dynamic correction of creatinine and D3‐creatinine using ultra‐performance liquid chromatography–tandem mass spectrometry

Xinyi Huang, Tianai Zhang, Xuan Wang, Shuqiao Wang, Manqing Nie, Xinyue Luo, Ming Yang, Xiaoli Zou
  • Clinical Biochemistry
  • Drug Discovery
  • Pharmacology
  • Molecular Biology
  • General Medicine
  • Biochemistry
  • Analytical Chemistry


This study developed a simple method for muscle mass determination based on D3‐creatine dilution by removing the matrix effects of ultra‐performance liquid chromatography–tandem mass spectrometry analysis through mutual correction of creatinine and D3‐creatinine. Rats were administered an oral tracer dose of D3‐creatine at age 6 weeks. Creatinine and D3‐creatinine in urine were detected using ultra‐performance liquid chromatography–tandem mass spectrometry after diluting 20 times to obtain D3‐creatinine enrichment factor (mole percent excess). The mole percent excess obtained from peak area could be used to calculate muscle mass using the improved formula. The limit of detection was 0.500 ng/mL for D3‐creatinine. Creatinine and D3‐creatinine could be mutually corrected because of the same matrix effect, and D3‐creatine spillage was negligible within 0.22%. Isotopic steady time was consistent with that obtained using conventional methods. Bland–Altman plots demonstrated the satisfying consistency between the proposed method and magnetic resonance imaging. This is a simple and rapid measuring method of muscle mass based on D3‐creatine dilution that requires no accurate quantification of creatinine and D3‐creatinine concentrations and no urine sample collection to obtain D3‐creatine spillage.

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