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Method for quantificationally detecting total amino acids of transfer factor capsule

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

Active Publication Date: 2018-11-16
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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  • Method for quantificationally detecting total amino acids of transfer factor capsule
  • Method for quantificationally detecting total amino acids of transfer factor capsule
  • Method for quantificationally detecting total amino acids of transfer factor capsule

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0062] In step 1), the preparation method of transfer factor capsule total amino acid content assay need testing solution, specifically comprises the following steps:

[0063] 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.

[0064] 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

[0086] The screening of embodiment 1 transfer factor capsule hydrolysis conditions

[0087] 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 3-factor 3 The horizontal orthogonal test table is shown in Table 1, and the analysis of the orthogonal test results is shown in Table 2:

[0088] Table 1 Orthogonal test table of transfer factor capsule hydrolysis condition screening

[0089] 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

[0090] Table 2 Analysis table of results of orthogonal test of transfer factor capsule hydrolysis condition screening

[0091]

[0092]

[0093] It can be seen from Table 2 that the o...

Embodiment 2

[0094] Example 2 The screening of chromatographic conditions for the determination of total amino acid content in transfer factor capsules

[0095] Three factors and three levels of orthogonal tests 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.

[0096] Table 3 Orthogonal test table for determination of total amino acid content in transfer factor capsules and screening of chromatographic conditions

[0097] Level

Moving Phase B Composition Ratio A

Column temperature B

Mobile phase flow rate C

1

98:2

25℃

...

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Abstract

The invention discloses a method for detecting total amino acid content of a transfer factor capsule. The method solves the technical problem that the prior art only detects free amino acids and cannot detect bonded amino acids of the polypeptide substance and a transfer factor is a multi-component biochemical drug in the amino acid detection process so that the separation degree of amino acid peaks and unknown component peaks is poor. Through a one-measurement multi-evaluation method, based on a glutamic acid reference substance, 18 common amino acids are quantitatively detected so that the technical problems of more reference materials, a high detection cost and complicated operation are solved. The method utilizes glutamic acid as an internal reference substance. Through the systematicverification research, relative correction factors and relative retention time of other 17 kinds of amino acids are determined and quantitative detection of 18 kinds of amino acids is realized. The method is accurate and reliable, has good separation effects and strong specificity and can effectively detect and monitor total amino acids in the transfer factor capsule.

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 Applications(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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