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3-Methlyhistidine Analysis

Heartland Assays provides 3-methylhistidine (3MH) analysis. Laboratory supervisor Dr. John A. Rathmacher has had extensive experience working with amino acids and specializes in the analysis of 3MH. Dr. Rathmacher has over 10-refereed publications and 20 abstracts relating to 3MH and its use in accessing muscle protein turnover. Heartland can analyze various samples, including those from lab animals, humans, cell cultures and tissues.


Urinary 3-methylhistidine

Urinary 3MH is important in the measurement of skeletal muscle (myofibrillar) protein breakdown. The procedure of measuring urinary 3MH entails a quantitative collection of urine, with the assumption that no metabolism of 3MH has occurred after being released from actin and myosin. If the precursor pool of 3MH (3MH bound to muscle protein) can be determined, then a fractional breakdown rate can be calculated; otherwise the measured amount of urinary 3MH can be used as an index of protein degradation.

 

 


 

Procedure

3MH will be quantified using an internal standard (ISTD) method. A dueterated-3MH molecule will be used for the procedure of the analysis and quantification as the ISTD. 3MH will be analyzed by gas chromatography/mass spectrometry in the laboratory using procedures developed by Dr. Rathmacher.1,2 All methods developed in the laboratory adhere to Good Laboratory Practices (GLP). These practices also utilize precision, accuracy, and quality control requirements that have been outlined in our procedures. All the data obtained during the procedure will be recorded in an Excel spreadsheet as a value of nmol/mL, unless the 24-h urine volume is provided prior to the analysis. If the customer provides this value, the results will instead be recorded as µmol/day.


Works Cited

  1. Rathmacher J.A., Link G.A., Flakoll P.J, and Nissen S.L. 1992. Gas chromatographic mass spectrometric analysis of stable isotopes of 3-methylhistidine in biological fluids: application to plasma kinetics in vivo. Biol. Mass Spectrom. 21: 560-566. View Abstract
  2. Rathmacher J.A., Flakoll P.J., and Nissen S.L. 1995. A compartmental model of 3-methylhistidine metabolism in humans. Am. J. Physiol. 269 32: E193-E198. View Abstract

 

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