Valicare's modern isolator-equipped test laboratory meets GMP requirements and the DIN EN ISO 14644 standard
We test the quality of your biological indicators, BIs, and support you with special laboratory services for process development and validation of your isolators.
Valicare's modern testing laboratory, the ValicareLab, is equipped with a sterilization isolator, meets the requirements of good manufacturing and laboratory practice, GMP and is in accordance with the ISO 14644 cleanroom standard.
The ValicareLab is a qualified testing laboratory in which comprehensive quality testing of biological indicators (BIs) used in hydrogen peroxide biodecontamination process development and validation is offered. The modern testing laboratory of Valicare s.r.o. is placed in Trenčianska Turná close to the city of Trenčín. It is designed to meet the requirements of good manufacturing and laboratory practice, GMP and is in accordance with the ISO 14644 cleanroom standard. The ValicareLab is equipped with a special edition of a Klenzaid ® STI isolator using hydrogen peroxide for decontamination and installed in an ISO Class 8 clean room. Klenzaids®, a joint venture member of Syntegon Technology GmbH, is a leading manufacturer of aseptic, bioclean and containment equipment in India (Mumbai).
Our experts, working in compliance with EMA or FDA standards and cGMP guidelines, provide high quality laboratory services, that follow the regulatory requirements and meet the specific requirements of the customers.
Laboratory activities are accomplished in compliance to our quality management system, which is built and sustainably improved in accordance to DIN EN ISO 9001:2015, and annually supervised by an independent certification body.
Our ValicareLab services include quality assurance of BIs as well as preparatory and supportive testing of customer-specific sterilization recipes and the performance of material studies.
Specifically, this includes the following tests:
- D-value determination for biological indicators (BIs) under defined standard conditions in accordance with the requirements of ISO 11138-7:2019
- D-value determination according to customer specific requirements (e.g. humidity, temperature, air flow)
- determination of the spore count and identification of the genus Geobacillus
- testing customized sterilization recipes (H2O2 concentration, humidity, temperature)
- preparation of D-value studies (material studies)
- testing of newly developed BIs
- development of bio decontamination process conditions for specific isolator's loading patterns
The D-value test determines and verifies the resistance of biological indicators (BIs) to the specified sterilization method according to ISO 11138: 2017 and according to the guidance for interpretation of results, ISO 11138-7:2019. This type of test might be also used to determine the approximate time of the biodecontamination processes.
Fast and reliable results that are documented in compliance with GMP are a matter of course for us.
Contact us in due time if we shall test your BIs in preparation for the process development and validation of your isolator, or if other special tests are needed.
We are ready and flexible to expand our offer within the scope of our possibilities.
Characterization of Biological Indicators for Decontamination Processes with vaporous hydrogen peroxide
The methods of spore counting and D-value determination are common practice for characterization and quality control of biological indicators
Sterilization processes preceding the aseptic filling of pharmaceuticals must be proven to be highly effective (EU GMP, Vol. 4, Annex 1). Biological indicators (BIs) are used to demonstrate the efficacy of decontamination or sterilization processes.
Both sterilization processes in autoclaves and decontamination processes using vaporous hydrogen peroxide (H2O2) in isolators usually use spores of Geobacillus stearothermophilus. These spores are an appropriate model system, as they are showing high resistance. Furthermore, Geobacillus stearothermophilus is cultivated at relatively high temperatures (55–60°C), which specifies the evaluation of the test.
BIs for decontamination processes consist of spore monolayers applied on stainless steel carriers. For better handling, these carriers are sealed in Tyvek® envelopes. Although this membrane is permeable for H2O2, it provides a diffusion barrier for the spore-inoculated carrier (worst case scenario).
After the application, carriers are transferred from the Tyvek® envelopes to nutrient media. If there is no growths after seven days of incubation, all spores on the carrier have been killed by the treatment process.
Transfer of spore carriers into nutrient medium
Decontamination processes are supposed to provide a 6-log reduction of microorganisms. Therefore, a BI must be occupied with at least 106 spores to be able to prove this.
Additionally, the spores should have a defined resistance. This resistance is prone to variations, since spores represent a non-calibratable biological system. The D-value (decimal reduction time) can be used to determine a characteristic value for resistance of a specific BI batch. Determined in one and the same system, D-values of different batches are comparable. Moreover, insights are gained about the killing behavior of a batch and possible hyper-resistant BIs („rogue BIs“, „late survivors“ or „anomalous BIs“). A batch containing such indicators should not be used for validations, as this may produce false positive results.
DIN EN ISO 11138-7:2019-11 defines the modalities to be followed for the determination of valid spore counts and D-values for characterization and quality control of BIs.
Our isolator experts can optionally support you in the following topics:
Spore count is determined according to DIN EN ISO 11138-7:2019-11: Sterilization of health care products – Biological indicators – Part 7: Guidance for the selection, use and interpretation of results (ISO 11138-7:2019); German version EN ISO 11138-7:2019 from four single samples using the heat shock method.
Determination of system D-value
The system D-value is determined following DIN EN ISO 11138-7:2019-11: Sterilization of health care products – Biological indicators – Part 7: Guidance for the selection, use and interpretation of results (ISO 11138-7:2019); German version EN ISO 11138-7:2019 from 30 sample groups, each with 10 or 20 single samples and a sampling interval of one minute.
The D-value is calculated according to the Limited Holcomb-Spearman-Karber Procedure (LHSKP) and is determined in a standardized system. Additionally customers can choose from a set of predefined values of H2O2 concentrations and relative humidities to adapt the D-value determination as closely as possible to its conditions.
Spore identification by API 50
Spore identification is performed by means of a bioMérieux API® 50 CHB/E test. In parallel, KWIK STIK™ of the strain to be identified is entrained.
Initial qualification and quality testing of the BIs is performed in the Valicare test laboratory using a pharmagrade sterilization isolator.
Contact us right in due time, please, if you plan to use BIs for process development and validation of your isolator system. With a preliminary lead time, we provide the necessary number of “qualified” ready-for-use BIs. In combination with a joint project we will gladly take over complete review and characterization of your batch of BIs or supplement missing partial aspects.
Take advantage of BIs tested and characterized under real conditions in an isolator. If you also use H2O2 for decontamination in your isolator, you will get the D-value for your BIs reliably determined in your application scope together with a statement on anomalies.
This saves you a lot of time during process development and validation.
Material Studies on the Biodecontamination Effect
The microbiological decontamination effect by hydrogen peroxide of surfaces varies strongly with the used material
In a study, "Suitability of different construction materials for use in aseptic processing environments decontaminated with gaseous hydrogen peroxide" conducted by experts from Syntegon Technology GmbH and published in 2007, 49 materials potentially used in aseptic processing environments were inoculated with spores of Geobacillus stearothermophilus ATCC®12980TM. In order to be able to record individual inactivation kinetics, the inoculated materials were exposed to reproducible hydrogen peroxide biocontamination processes for defined periods of time and the D-values were repeatedly determined.
It was found that both the chemical composition of the material and the nature of the surfaces effected the resistance of the spores. The calculated D-values show some correlation to the roughness as well as the wettability of the materials. The full article can be found here.
Valicare offers such kind of material studies to customers. Materials inoculated with spores of Geobacillus stearothermophilus are exposed to reproducible biodecontamination processes with hydrogen peroxide in Valicare's own isolator over defined periods of time. The determination of the D-value is used as a measure of the material-dependent resistance of the spores.
Contact our experts if you have any questions about the procedure or would like to know more about our material studies.