DETERMINATION OF AMINO ACIDS IN THE UNOSSIFIED ANTLERS OF MARAL (CERVUS ELAPHUS): VALIDATION OF A PROCEDURE

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DOI: https://doi.org/None
Issue: 
4
Year: 
2016

N.P. Zemtsova, Professor V.F. Turetskova, PhD; O.G. Makarova, PhD Altai State Medical University; 49, Lenin Pr., Barnaul 656038, Russian Federation

Background. Amino acids that are due to their adaptogenic activity are one of the extensive and stable groups of biologically active substances in the unossified antlers of maral (Cervus elaphus). A procedure [4] using high performance liquid chromatography (HPLC) has been proposed to assess the quality of unossified maral antlers. Objective: to estimate the validation of the developed HPLC procedure for quantifying essential amino acids (glycine, alanine, and proline) in the crushed unossified maral antlers. Material and methods. Crushed unossified maral antlers stored during the planned cutting of unossified stag antlers in May 2014 were the subject of the investigation. The raw material was crushed with a MSH-100 ball grinder. The hydrolysate was extracted with 6M hydrochloric acid, followed by amino acid modification with phenyl isothiocyanate and further separation of phenylthiocarbamil derivatives of amino acids. The investigations were performed on a LC-20 Prominence liquid chromatograph (Japan) using an UV detector. Results. The validation of the procedure was assessed using the following characteristics: specificity, linearity, suitability, correctness, and precision during repeatability. It was found that the presence of accessory agents did not affect the results of the analysis. Spectral ratios, retention times, and absorption maxima of the substances analyzed in the test and standard samples were within the acceptance limits. The investigation revealed that the procedure was proper and suitable during repeatability, as well as a linear relationship of the magnitude of an analytical signal to the amount of a detectable substance. Conclusion. The procedure for HPLC quantification of amino acids, such as glycine, alanine, and proline, is valid in specificity, suitability, correctness, linearity, and precision during repeatability and may be recommended to assess the quality of crushed unossified maral antlers.
Keywords: 
unossified maral antlers
validation
amino acids
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