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Identification method of red nocardia cell wall skeleton

A technology of Nocardia rubra and cell wall, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems such as the difficulty of distinguishing components, and achieve the effect of easy separation, good separation effect and strong reliability

Active Publication Date: 2014-10-22
FUJIAN SHANHE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a new method for identifying the cell wall skeleton of Nocardia rubra, so as to solve the problem that the components in the prior art are difficult to distinguish

Method used

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  • Identification method of red nocardia cell wall skeleton
  • Identification method of red nocardia cell wall skeleton
  • Identification method of red nocardia cell wall skeleton

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Spotting

[0040] Use a 2B pencil to draw a straight line at 1.0 cm from the bottom edge of the thin-layer plate as the sampling baseline, and draw 3 dots on the sampling baseline at a distance of 1.0 cm, and respectively apply the reference solution and the test solution. Take 2 μl each of the alanine reference solution and the test solution, and spot them on the thin-layer plate. Take 5 μl each of the mesodiaminopimelic acid reference solution and the test solution, and spot them on a thin-layer plate.

[0041] (2) expand

[0042] Put the thin-layer plates of the alanine reference substance solution and the test solution into the n-butanol-glacial acetic acid-water (8:3:1) developer, and place the meso-diaminopimelic acid Put the TLC plates of the reference substance solution and the test solution into the developing agent of sodium lauryl sulfate-n-butanol-n-hexane-water (6:25:6:20). The depth of immersion in the developer is 0.5cm from the bottom of the thin-...

Embodiment 2

[0048] (1) Spotting

[0049] Use a 2B pencil to draw a straight line at 1.0 cm from the bottom edge of the thin-layer plate as the sampling baseline, and draw 3 dots on the sampling baseline at a distance of 2.0 cm, and respectively apply the reference solution and the test solution. Take 2 μl each of the alanine reference solution and the test solution, and spot them on the thin-layer plate. Take 5 μl each of the mesodiaminopimelic acid reference solution and the test solution, and spot them on a thin-layer plate.

[0050] (2) expand

[0051] Put the thin-layer plates of the alanine reference substance solution and the test solution into the n-butanol-glacial acetic acid-water (8:3:1) developer, and place the meso-diaminopimelic acid Put the TLC plates of the reference substance solution and the test solution into the developing agent of sodium lauryl sulfate-n-butanol-n-hexane-water (6:25:6:20). The depth of immersion in the developer is 1.0cm from the bottom of the thin-...

Embodiment 3

[0057] (1) Spotting

[0058] Use a 2B pencil to draw a straight line at 1.0 cm from the bottom edge of the thin-layer plate as the sampling baseline, and draw 3 dots on the sampling baseline at a distance of 1.5 cm, and respectively apply the reference solution and the test solution. Take 2 μl each of the alanine reference solution and the test solution, and spot them on the thin-layer plate. Take 5 μl each of the mesodiaminopimelic acid reference solution and the test solution, and spot them on a thin-layer plate.

[0059] (2) expand

[0060] Put the thin-layer plate of the alanine reference substance solution and the test solution into the n-butanol-glacial acetic acid-water (8:3:1) developer, and place the meso-diaminopimelic acid Put the TLC plates of the reference substance solution and the test solution into the developing agent of sodium lauryl sulfate-n-butanol-n-hexane-water (6:25:6:20). The depth of immersion in the developer is about 1.0cm from the bottom of the ...

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Abstract

The invention discloses an identification method of red norcardia cell wall skeleton. The method comprises the following steps: (1) sample spotting: using at least two silica-gel G thin-layer plates, spotting a proper amount of an alanine reference solution and a test sample solution on the first thin-layer plate; then spotting a proper amount of a meso-diaminopimelic acid reference solution and a test sample solution on the second thin-layer plate; (2) development: soaking the first thin-layer plate into a developing solvent composed of n-butanol-glacial acetic acid-water according to a volume ratio of 8:3:1, soaking the second thin-layer plate into a developing solvent composed of sodium dodecyl sulfate-n-butanol-n-hexane-water according to a volume ratio of 6:25:6:20; wherein the distance between the surface of the developing solvent to the bottom edge of the thin layer plate is in a range of 0.5 to 1.0 cm and the sample spots cannot be emerged by the developing solvent; sealing the top cover, when the developing solvent reaches the two thirds of the total length of the thin-layer plate, taking out the thin-layer plate, drying; (3) color developing; (4) observation. The identification method has the advantages of clear color development, high separation degree, and high reliability.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a new method for identifying the cell wall skeleton of Nocardia rubrum. Background technique [0002] Nocardia rubra (Nocardia rubra) is an immunologically active microorganism. Its cell wall skeleton is obtained through cell crushing, removal of cell contents, proteins and lipids and other components. [0003] The cell wall skeleton of Nocardia rubrum is an active substance containing nocardia mycolic acid, neutral polysaccharides and mucopeptides, which can stimulate cellular immune function, promote the secretion of various cytokines, and participate in the process of anti-tumor immunity. Treatment or adjuvant therapy of various tumors. [0004] It needs to be identified when producing the Nocardia rubra cell wall skeleton. The identification method of prior art is as follows: to the qualitative analysis of cell wall characteristic aminoacid mesodiaminopimelic acid, its method is...

Claims

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Application Information

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IPC IPC(8): G01N30/90
Inventor 徐镜钟丽芳查贝贝谢必峰金勇邵恩文
Owner FUJIAN SHANHE PHARMA
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