1. Introduction
Vancomycin is a water-soluble protein made from recombinant genes. A recombinant protein based on the chemical structure of glycopeptide drugs (vancomycin, teicoplanin). The active center of its gene fragment can destroy the structure of many amide bonds in the structural formula of glycopeptides, and lose the antibacterial property after ring-opening.
2. Application
It can inactivate the antibacterial properties of vancomycin for injection, teicoplanin and oral preparations, so that the positive bacteria in stereosis test can grow well. The recovery rate of positive bacteria in the microbial limit test is greater than 70% to ensure the accuracy of the test work.
3. Operation
3.1 Sterility test
For each batch of vancomycin and teicoranin samples, add 1 bottle of vancomycin enzyme (5ml/ bottle) according to the following table:
Add 1ml of enzyme to each of the 5# and 6# rinsing solution, shake well, first moisten the filter membrane of the incubator with the 5# rinsing solution, pump the dissolved sample solution through the incubator, and successively rinse the filter membrane of the incubator with a small amount of rinsing solution (1#~6#) several times. Try not to accumulate liquid on the filter membrane, and finally pump 100ml of the corresponding medium into the incubator, then add 1ml of vancomycin, and shake well.
3.2 Microbiological limit test
Add 4.5ml vancomycin enzyme to 1000ml peptone rinse and shake well. First wet the filter membrane about 50ml, the dissolved sample solution through the filter membrane in turn with the peptone containing vancomycin enzyme rinse, a small amount of multiple times, as far as possible on the filter membrane without fluid accumulation. Remove the filter membrane and put it on the medium plate. Moisten the whole filter membrane with 0.5ml vancomycin enzyme for 20 minutes, let the enzyme directly inactivate the trace glycopeptides remaining on the filter membrane, and calculate the recovery rate.
3.3 Verification
Before the work of 3.1 and 3.2, the quality department of the user unit shall carry out the verification work and save the verification data. Write a standard SOP for validation based on the results.
1. Inactivation test
To remove the antibacterial properties of antibiotics, it can be called neutralization and inactivation.
Take 20mg/ml of vancomycin and 1ml of teicoranin solution, put it in a sterilized empty test tube, add 1ml of vancomycin enzyme, leave it at room temperature for 20 minutes, add 25ml of buttermilk soy peptone medium, shake, then add 1ml of Staphylococcus aureus (10 minus 6 or 100CFu/ml), shake well. Culture at 37℃ for 5 days, shake the test tube every day, and observe the growth of bacteria.
Vancomycin enzyme without adding 1ml was also taken as a negative control.
The positive tube bacteria should grow well within 5 days, and the negative tube should grow sterile.
2. Regulatory requirements
According to the provisions of the Chinese Pharmacopoeia, the test method needs to be validated by methodology, and the antibiotic drugs can be neutralized and their antibacterial properties removed to ensure the feasibility of the test method and the accuracy of the test results. Vancomycin enzyme is prepared according to the principles of sterility test in the pharmacopoeia appendix and the actual characteristics of antibiotics. Drugs that target glycopeptides are effective. Each pharmaceutical unit combined with glycopeptide drug products for methodological verification, in full compliance with regulatory requirements.
3. Packaging specifications of the product
Vancomycin is a sterile aqueous protein packaged in cillin bottles, rubber plugs and aluminum caps. 3 million U/ bottle.
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