BioLumix rapid microbiological testing meets rules that save time and cost

BioLumix rapid microbiological testing meets rules that save time and cost

Author: Ruth Eden (United States),

Tracy Yuan (formerly, Shanghai Meikai Pure Biotechnology Co., Ltd.)

Summary

Rapid microbiological testing methods (RMM's) are receiving increasing attention due to their high degree of automation, significantly reduced reporting time, ease of operation, no need for extensive training for employees, faster product release and improved process control. . However, three conditions must be met before using the rapid detection method: (1) comply with GMP rules; (2) verify with standard methods (such as the national standard method); (3) ensure that the investment can be rewarded. This article describes how to achieve the above three conditions and the advantages of RMM.

rule

Since June 25, 2010, the United States has required all nutraceutical companies to comply with the US Food and Drug Administration (FDA) cGMP rules, whether they are large companies with more than 1,000 employees or small businesses with only a small number of employees. . One of the most challenging provisions of cGMP is the 111.310 chapter. This section stipulates that each manufacturer must perform any necessary tests and inspections with “appropriate laboratory facilities to determine: a) whether the ingredients used are in compliance; b) whether the process specifications comply with the provisions of the main production records; c) dietary additives Production meets the requirements. This means that the manufacturer needs to take measures to ensure that the product is free from harmful microbial contamination. To do this, it is necessary to detect the relevant microorganisms from raw materials, intermediate products to end products.

Strengthen quality control to comply with cGMP rules

Traditional flat-panel methods that require long incubation times, trained microbiology experimenters, and cumbersome procedures have become bottlenecks in product release; simpler alternative detection methods such as rapid microbiological assays (RMM's) can Solve these problems well. When evaluating an alternative test method, there are three key issues to consider: (I) Has the system been verified? (II) Is it in compliance with GMP regulations? (III) What is the return on investment (ROI)?

Verification requirements

In the United States, the nutraceuticals and dietary additives industry should test their products in accordance with the guidelines in the USP <2021>, <2022> and <2023> sections. Guidance on Determining Appropriate Tests In Section <2023>, the actual qualitative and quantitative test procedures are located in sections <2021> and <2022>. All methods of replacing USP plates such as RMM's must be validated in accordance with USP guidelines. The verification of a microbiological detection method is an experimentally determined process that meets the needs of the application compared to conventional methods. The new Chinese Pharmacopoeia, which was implemented in October 2010, also gives the “Guidelines for the Verification of Alternative Methods for Microbial Rapid Testing”, which clearly indicates that companies can adopt rapid microbiological testing methods after relevant verification.

What are the verification points?

System verification includes installation qualifications, operational qualifications, and operational qualifications, and new system results must be ensured to be reliable and consistent with USP methods.

The first part: Installation Qualification ( IQ ) includes major component validation and verification, verification of environmental conditions, power requirements, computer qualifications, installation checklists and calibration files.

The second part: Operational Qualification ( OQ ) includes a unique standard operating procedure (SOP) for product analysis and operation, training documents, software features, compliance with 21 CFR Part 11 rules, and an operational checklist.

The third and most important part is the running qualification ( PQ ) , the core of the verification package. The components of operational eligibility include: parallel contrast, specificity, detection limits, reproducibility, durability, stability, false negative rate, and false positive rate.

BioLumix system has been verified to comply with cGMP BioLumix microbial real-time fluorescence photodetection system is a RMM designed specifically for the nutrition and health care products, food, cosmetics, pharmaceutical, water treatment and other industries, has been verified to comply with cGMP, the following is about the BioLumix system verification work A brief description.

1. Parallel comparison test between Biolumix system and USP method: Each comparative test was tested with more than 200 clean and naturally contaminated nutraceuticals and dietary supplement samples. The overall coincidence rate between USP and Biolumix methods was: bacteria The total number was 99.5%, the mold and yeast count was 97.9%, the Enterobacteriaceae count was 98.7%, the coliform count was 98.6%, E. coli was 99.4%, and Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella were both 100%.

2. Specificity (also known as compatibility and repellency): Includes testing each test tube to ensure that each strain can be detected. The results showed that each of the test tubes exhibited excellent compatibility and repellency.

3. The limit of detection is defined as the minimum level of microorganisms detectable by this method. All samples in the first trial were inoculated at 1.2 cfu/g and the second trial was inoculated at 1.5 cfu/g. The results show that the detection limit of the BioLumix system is equal to or higher than the plate count method , which is close to the ability to detect the growth of a bacterium in the medium. In many cases, one strain can be detected in the BioLumix test tube and the plate method cannot be detected; 20 samples of bacteria are detected in the Bio Lumix system in samples of 1-2 CFU levels, and the plate counting method is used. Only 12 samples were detected, and the data is shown in the table below.

4. Reproducibility: Refers to samples of natural contamination and samples inoculated with various strains to check the standard deviation (SD) of each medium. The following four graphs show that the method has good reproducibility.

5. Durability: including checking the performance of the system when the test parameters are slightly changed. For the BioLumix system, the parameters include the influence of the incubation temperature change of the instrument, the influence of the sample size, and the influence of the medium volume in the test tube. The durability is different. The medium was tested repeatedly. The results demonstrate that the system is extremely durable and unintentionally subject to small changes in method parameters or under a variety of different conditions.

6. Stability: Test the accuracy of the system when faced with the challenges of multiple detection conditions. That is, the accuracy of the test results obtained by analyzing the same sample under different normal test conditions, such as different analysts, instruments, or reagent lot numbers. The BioLumix system exhibits excellent stability and results in highly accurate results.

7. False negative: refers to a sample that is inoculated or naturally contaminated as a positive result in the reference method (plate count) and negative in the alternative method (BioLumix). The BioLu mix system showed no false negatives when performing bacterial counts and mold yeast tests and the USP plate count method had a false negative.

8. False positive: It means that a sample that has not been inoculated with the target bacteria has a positive result in the system and a negative result in the plate counting method. In the test of the four types of samples, the bacterial total bacteria test showed a 2.5% false positive rate in the BioLumix system, and no false positives were found in the mold and yeast tests.

The above verification process indicates that BioLumix will provide customers with a complete set of verification procedures by providing IQ and QQ files and parallel comparison tests after the customer selects the BioLumix system.

Return on investment ( ROI )

More and more companies are finding it cumbersome and time-consuming to use traditional microbiological testing or to send samples to an external laboratory for microbiological testing, and not necessarily to save money. In contrast, the advantage of using rapid microbiological assays (RMM's) is that the time to report test results can be reduced from 7-10 days to 5-48 hours. This allows companies to react more quickly to pollution, speed up product release, get products to market faster, save storage space and enhance product quality. The advantages of RMM include higher accuracy, better sensitivity, increased sample throughput, automatic data collection and more convenient data processing, and reduced product release costs.

The cost of purchasing a BioLumix system primarily consists of fixed asset investments and the use of disposable test tubes. Although the price of a disposable test tube is higher than that of a normal agar plate, since the BioLumix system does not require gradient dilution, only one test tube is required to test a sample, and the plate count method usually requires 4-6 plates. . In addition, the potential savings of rapid inspection systems include: reduced product release time, no need for excessive inspectors, lower reprocessing costs, reduced plant downtime, reduced inventory time for raw materials, intermediate products and end products, Reduce the chances of recall, increase the flexibility of products, etc.; thus better enhance and protect the company's image.