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A method for quantitative detection of total amino acids in transfer factor capsules

A quantitative detection method, the technology of total amino acids, applied in the field of quantitative detection of total amino acids in transfer factor capsules, can solve the problems of deviation of detection results, high detection cost, cumbersome operation, etc., to improve detection accuracy, reduce overlapping ratio, and realize supervision The effect of control

Active Publication Date: 2021-06-29
XIAN DAQING PHARMA FACTORY JINHUA ENTERPRISE GROUP CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the existing technology only detects the content of free amino acids in transfer factor, and cannot detect the bonded amino acids that make up polypeptides; A large degree of overlap results in a large deviation in the detection results
At the same time, the existing technology uses the external standard method for quantitative detection, which requires 18 kinds of amino acid reference substances, the detection cost is high, and the operation is cumbersome

Method used

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  • A method for quantitative detection of total amino acids in transfer factor capsules
  • A method for quantitative detection of total amino acids in transfer factor capsules
  • A method for quantitative detection of total amino acids in transfer factor capsules

Examples

Experimental program
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Effect test

preparation example Construction

[0063] In step 1), the preparation method of transfer factor capsule total amino acid content assay need testing solution, specifically comprises the following steps:

[0064] a. Accurately weigh about 1.0 g of the contents of the transfer factor capsule, put it in a 20ml stoppered test tube, add 0.1mol / L hydrochloric acid solution, shake on an oscillator to dissolve the transfer factor, shake well, and let stand; take the supernatant for use Filter through a 0.45 μm microporous membrane, accurately measure 2.0 mL of the subsequent filtrate, put it in a digestion tube, add 6 mL of concentrated hydrochloric acid, seal it with nitrogen, place it at 110 ° C for 24 hours, and let it cool to prepare the transfer factor acid hydrolyzate.

[0065] b. Unseal the transfer factor acid hydrolyzate, put it in an evaporating dish, and evaporate to dryness in a water bath; add 10 mL of 0.1 mol / L hydrochloric acid solution to dissolve the residue, and use it as the test solution for the deter...

Embodiment 1

[0087] The screening of embodiment 1 transfer factor capsule hydrolysis conditions

[0088] The hydrolysis conditions of transfer factor capsules were screened by 3-factor 3-level orthogonal test. The 3 factors investigated for hydrolysis conditions were hydrolyzed sample concentration, hydrolyzed time, and hydrolyzed acid volume; The horizontal orthogonal test table is shown in Table 1, and the analysis of the orthogonal test results is shown in Table 2:

[0089] Table 1 Orthogonal test table of transfer factor capsule hydrolysis condition screening

[0090] Level Hydrolysis sample concentration A Hydrolysis time B Hydrolyzed acid volume C 1 2.0mg / ml 22h 3mL 2 2.4mg / ml 24h 6mL 3 2.8mg / ml 26h 9mL

[0091] Table 2 Analysis table of results of orthogonal test of transfer factor capsule hydrolysis condition screening

[0092]

[0093]

[0094] It can be seen from Table 2 that the optimal combination of hydrolysis conditions d...

Embodiment 2

[0095] Example 2 The screening of chromatographic conditions for the determination of total amino acid content in transfer factor capsules

[0096] Three factors and three levels of orthogonal test were used to screen the total amino acid content of transfer factor capsules and determine the chromatographic conditions. The three factors investigated in the chromatographic conditions were the composition ratio of mobile phase B, the column temperature of the chromatographic column, and the flow rate of the mobile phase; See Table 3 for the 3-factor 3-level orthogonal test table of the optimal level value design, and see Table 4 for the analysis of the results of the orthogonal test.

[0097] Table 3 Orthogonal test table for determination of total amino acid content in transfer factor capsules and screening of chromatographic conditions

[0098] Level Moving Phase B Composition Ratio A Column temperature B Mobile phase flow rate C 1 98:2 25℃ 0.8mL / min ...

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Abstract

A method for detecting the total amino acid content in transfer factor capsules disclosed by the present invention solves the problem that in the prior art, only free amino acids are detected, but the bonded amino acids constituting polypeptide substances cannot be detected, and the problem of transfer factor For multi-component biochemical drugs, the technical problem of poor separation between each amino acid peak and the unknown component peak is adopted. At the same time, the "one test and multiple evaluation" method is adopted to realize the quantitative detection of 18 common amino acids through a glutamic acid reference substance, which solves the problem The external standard method needs more reference substances, the detection cost is high, and the technical problems of cumbersome operation. The invention uses glutamic acid as an internal reference, and through systematic verification research, determines the relative correction factors and relative retention times of other 17 kinds of amino acids, and realizes the quantitative detection of 18 kinds of amino acids. The detection method is accurate and reliable, has good separation effect and strong specificity, and can effectively detect and monitor the total amino acids in the transfer factor capsules.

Description

technical field [0001] The invention belongs to the technical analysis field of multi-component biochemical medicines, and in particular relates to a quantitative detection method for total amino acids of transfer factor capsules. Background technique [0002] Transfer factor (transter factor, referred to as TF), also known as transfer factor, is a natural two-way immunomodulator, and is currently an ideal drug for the treatment of immunocompromised, deficient and other related diseases. Transfer factor is a polypeptide substance, which is a composite molecule composed of polynucleotides, polypeptides and other components. Among them, the small molecule peptides are the functional markers of the transfer factor and serve as its main functional components. Amino acid is the basic unit of polypeptide and the key quality index of transfer factor. Therefore, accurate determination of the total amino acid content in transfer factor is of great significance for optimizing the pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 李小凤蔡慧侠郭维真杨静刘福敏
Owner XIAN DAQING PHARMA FACTORY JINHUA ENTERPRISE GROUP CORP
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