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Buffer liquid for bacteria endotoxin horseshoe crab test detection method and process for preparing same

A technology of bacterial endotoxin and buffer solution, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve problems such as complicated, difficult to control accurately, and endotoxin pollution

Inactive Publication Date: 2005-06-01
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This process is very complicated and difficult to control accurately, especially the contamination of endotoxin is easy to introduce, so there is an urgent need for a buffer that can be used to dilute the sample and at the same time adjust and indicate the pH of the sample

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Example 1. Standard curve for endotoxin determination

[0010] On the endotoxin analyzer, based on the standard curve LogT (OD0.02) =a+b Log(c), use the standard endotoxin concentration of 0.22-100EU / ml to test with the Limulus reagent with a sensitivity of 0.06EU / ml, and obtain the corresponding standard curve as LogT (OD0.02) =-0.4129Log(c)+3.2038, where T (OD0.02) is the characteristic reaction time, and c is the endotoxin concentration.

Embodiment 2

[0011] Embodiment 2. Preparation 1 of Tris-HCl damping solution containing phenol red

[0012] Weigh 0.005 g of phenol red and 0.363 g of Tris salt, place them in a clean glass bottle, and dry-bake them at 200 degrees for 1 hour to eliminate endotoxins; add 100 ml of endotoxin-free water, and adjust the pH to 7.0, high-pressure sterilized aseptic storage or frozen storage for later use.

Embodiment 3

[0013] Embodiment 3. the preparation 2 of the Tris-HCl buffer solution containing phenol red

[0014] Weigh 0.1 g of phenol red and 6.05 g of Tris salt and put them in a glass bottle, dry-bake at 150 degrees for 5 hours to eliminate the endotoxin; add 100 ml of endotoxin-free water, adjust the pH to 8.5 with concentrated hydrochloric acid, Autoclaved sterile storage or frozen storage for later use.

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Abstract

The buffering liquid for limulus test for detecting bacterian endotoxin is Tris-HCl buffering liquid containing phenol red in 0.005-0.1 wt% concentration. The preparation process includes weighing phenol red and Tris salt and stoving at 150-200 deg.c for 1-5 hr; adding no-endotoxin water and regulating pH to required value with concentrated hydrochloric acid; high pressure sterilization and no-bacteria preservation or freezing preservation. The buffering liquid is used in diluting tested sample and regulating and indicating sample pH value. The buffering liquid for limulus test for detecting bacterian endotoxin has no influence on the endotoxin in the detection of two kinds of endotoxin and coincides with the requirement for limulus test.

Description

Technical field: [0001] The invention relates to a limulus test detection method of bacterial endotoxin, and in particular provides a buffer solution for regulating and indicating pH of a sample in the limulus test. Background technique: [0002] Bacterial endotoxin (Bacterial Endotoxin) is a macromolecular substance with various biological activities produced by Gram-negative bacteria, and its main chemical component is lipopolysaccharide (LPS). Endotoxin is the main pollutant in injection drugs (raw materials, etc.). [0003] At present, the detection methods of bacterial endotoxin mainly include rabbit test method and limulus test method. The limulus test is an in vitro detection method that uses the mechanism of agglutination reaction between the limulus reagent and endotoxin to detect the bacterial endotoxin infected in the blood of the drug and the body in a qualitative (gel method) or quantitative (dynamic turbidity method) method. . Normally, within a certain pH r...

Claims

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Application Information

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IPC IPC(8): G01N33/579
Inventor 李京华邵英光王俊德
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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