Method for determination of free amino acid in spleen aminopeptide

Abstract

The invention discloses a method for determination of free amino acid in spleen aminopeptide, comprising the following steps: Step 1, solution preparation: a reference stock solution, an internal standard stock solution, a mobile phase A and a mobile phase B are prepared; Step 2, sample treatment; Step 3, precolumn derivatization; Step 4, chromatographic separation, sample introduction, gradient elution, detection wavelength being 338 nm and 262 nm respectively, and determination of peak area; and Step 5, free amino acid content calculation. The invention has the following advantages and effects: by liquid chromatogram of precolumn derivatization, a sample undergoes derivatization and then sample introduction is carried out, and through gradient elution during chromatographic separation, free amino acid in spleen aminopeptide is successively eluted; through selection of an amino acid standard substance for chromatographic elution, equation of linear regression is calculated by linear regression of solution mass concentration X to peak area Y, and content of free amino acid in spleen aminopeptide is calculated according to the peak area. Accuracy of determination results is high, and it is convenient to control the content of a spleen aminopeptide product.

Description

technical field [0001] The invention relates to the technical field of biological products, in particular to a method for measuring free amino acids in spleen aminopeptide. Background technique [0002] Spleen aminopeptide is a biochemical drug extracted from animal organs. Its main components include amino acids, polypeptides, nucleotides and other substances. It is used as an immune regulator for recurrent respiratory tract infections, bronchitis, asthma, pneumonia, cancer, etc. adjuvant treatment of disease. Clinical observation shows that it can effectively improve the resistance of patients, reduce the incidence of respiratory tract infection, and has the function of regulating cellular immunity. The main active substance in spleen aminopeptide is polypeptide, and the national drug standard WS1-(X-002)-2002Z Among them, the determination of the content of polypeptide is the only index to control the content of spleen amino peptide products, and there is no measurement ...

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Problems solved by technology

However, during the determination of the above-mentioned free amino acids, during the degradation and destruction process of the spleen amino peptide oral liquid under forced conditions, the polypeptides in the spleen amino peptide are easily degraded to form amino acids, and the phenomenon pointed out by Zuo Guangzhi and others in the literature appears, that is, in the determination The content of free amino acids will increase, which will affect the accuracy of the determination results

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