This internship report titled “Endotoxin Testing and Analysis of the Environmental Monitoring of Quality Control in Kalbio Global Medika” has been written to fulfill the requirement to complete this credited internship task. I am grateful for the opportunity to do an internship at PT Kalbio Global Medika, one of the premier biopharmaceutical companies in Indonesia, where I met many wonderful and wonderful people. Finally, special thanks to my fellow internship friends from PH19, Celine Chelovna, Janice Evita Sundah and Putri Avanny Jamaica, who became my companion during my internship at KGM and shared all the new experiences together.
Kalbio Global Medika (KGM), which is a subsidiary of PT Kalba Farma, is one of the primary pharmaceutical companies in Indonesia specializing in the production of sterile parenteral drugs, which was established in 2014. Endotoxin testing and environmental monitoring (EM) analysis are two of the most important tests performed in the microbiological QC department. The result obtained from the endotoxin testing of the water for injection (WFI) sample, erythropoietin medium and buffer, ephepoetin medium and buffer, and raw material citric acid monohydrate shows a successful result with no clot in the sample and the presence of clot in the product positive control ( PPC).
Kalbio Global Medika (KGM) is one of the primary and the pioneers of biopharmaceuticals in Indonesia, built in 2014 and authoritatively launched in 2018 by the President of the Republic of Indonesia, Ir.
Description of Department
Product of the Host Institution
Placing insulin glargine next to the freezer compartment or freezer pack is not recommended and the pre-filled pack should be kept in the outer carton in order to protect from light. An opened pre-filled Donkey, whether stored or not refrigerated, should be used within 28 days, protected from heat and light. If the pre-filled pack has not been refrigerated, an opened and used pre-filled pack can be stored unrefrigerated and away from heat and direct light for up to 28 days, as long as the temperature does not exceed 25°C.
Leucogen is also one of the Kalbio products where the formulation of this drug is Filgrastim as seen in figure 4 below. Regarding indications, Leucogen is indicated for shortening the period of neutropenia in patients with solid tumor cancer or non-myeloid malignancies receiving myelosuppressive cytotoxic chemotherapy .
PROJECT DESCRIPTION 2.1 Internship Project
- Project background
- Scope of the project
- Objective
- Problem formulation and proposed solution 1 Problem Formulation
- Methodology for Endotoxin Testing
- Methodology for Environmental Monitoring
Before starting the endotoxin test, pH measurements of the sample are taken to ensure that the pH of the sample is within an acceptable range of 6 to 8. If the sample pH results are out of range, adjust prior to testing by adding NaOH or HCl solution accordingly. Thereafter, the maximum dilution value (MVD) for each of the samples is determined by looking at the information for endotoxin limit, sample concentration and TAL sensitivity of the incoming sample and the sample dilution for BET on google sheet.
Each sample needs a sample replicate and a positive product control (PPC) replicate, which consists of sample and CSE. For sample that was diluted, 200 μL of sample from the corning or pyrogen-free tube was added to the TAL, and for the sample with dilution, 100 μL of the diluted sample from the pyrogen-free tube/depyrogen-free tube was added to the TAL ampoule bottle previously water has been added to the BET. The final concentration of the diluted CSE should be 4 times λ (lambda/TAL sensitivity).
After the diluted CSE was vortexed, 100 μL of diluted CSE was added to the TAL for PPC (positive product control) and positive control. After the required amount of incubation time is met, the media needs are extracted and the result is read and written to each of the respective formats. On the last day of the sterility test or on the 14th day, a photograph is taken of the box used in the sterility test.
The sample's request for analysis (RFA), date and signature of the personnel assigned to each of the actions are included in the logbook. From the RFA number of the sample and information written on the RFA form, identify what grade the media is. This was done by separating the OOS media from the rest of the media, it in the.
A picture of the OOS media is taken and printed out to be attached to the OOS form. The OOS media are then parafilmed and stored in the refrigerator (RFR002) for further identification of the OOS result by assigned personnel.
FINDINGS 3.1. Result
Endotoxin Testing
Numbers one through five in Figure 11 indicate the two samples of medium and buffer for erythropoietin and the other two samples of medium and buffer for ephepoietin with the number five as positive and negative controls on the left and right, respectively. As for numbers six and seven in Figure 13, those two show the result of citric acid monohydrate and the control for sample with TAL sensitivity of 0.25 EU/ml. The controls from each group are used as a comparison to determine whether or not the endotoxin tests have been passed.
Confirmation of the endotoxin test result was made by the presence of a gel clot. It can be considered passed when both duplicate samples (on the right) show negative results as negative control. In addition to the samples, the PPC should show the presence of clot as well as the positive control showing its differences with the sample when endotoxin agents were present through the addition of CSE.
Figures 11, 12 and 13 mainly show successful results of endotoxin testing for all samples by the presence of clot in the PPC and absence of clot in the sample. However, for the endotoxin result of the raw material sample of citric acid monohydrate, as shown in number 6 of Figure 13, this was done in its MVD. This means that the previous tests performed with the final dilution in TAL reagent failed with no clot in the PPC, as shown by the third TAL ampoule bottle from above in Figure 14 above.
Therefore, the endotoxin test had to be re-tested with his MVD to make sure there was no endotoxin. As a result, citric acid monohydrate endotoxin testing is repeated using its MVD and the successful result with the presence of clot in the PPC and the absence of clot in the sample. Based on the citric acid monohydrate endotoxin test result using its MVD, it is now safe to say that the sample is free of endotoxin agents with no clot formation in the TAL on the right (for sample) and clot formation in the TAL on the left (for PPC).
Environmental Monitoring
The colony count result from the plate media exceeded the acceptance limit for Class D cleanroom, which is <200 CFU/m3. Therefore, it is considered a failure and an OOS forms investigation should be established to further investigate the root cause of the OOS failure. Further investigation of the OOS failure is done by QC compliance personnel along with the personnel who did the EM.
Apart from colony plate reading, sterility observation of KGM finished good product results are obtained every 14 days or after a complete incubation period or sterility testing has been done. Results obtained from sterility testing observation photos of Hemapo 3000 IU/mL sample with its respective control TSB or FTM media placed side by side as can be seen from figure 17 below. No differences in color and turbidity of the sample with its control media demonstrates the success of sterility testing of samples performed by other QC personnel.
Sterility test results of the finished good Hemapo 3000 IU/mL can be seen in the picture below.
Analysis/Discussion .1 Endotoxin Testing
- Environmental Monitoring
Therefore, as mentioned above in the results section, a sample without endotoxin should have no clot formation and should be considered negative (Hashmi & Thakur, 2019). Based on the sample in Figures 5, 6 and 7 above, a successful endotoxin test result is evident from the absence of a clot in the sample and the presence of a clot in the PPC, demonstrating that the samples are endotoxin free. With each appropriate final sample dilution used in the TAL reagent, the image above shows a successful endotoxin test result.
However, with the raw material sample of citric acid monohydrate as mentioned in the results section above, it underwent retesting with its MVD. However, when no clot is formed in PPC or clot is still formed in the sample TAL, we can conclude that it has failed and endotoxin agents are present. In addition to that, staff may make certain errors and mistakes throughout the process which will affect the final result of the test (Sharma et.al., 2011).
In the production of pharmaceutical and biological products, the environment plays an important role in microbial access that can affect the products' quality, depending on how the environmental controls are carried out specifically in the production area. With an objection to establish a controlled environment systems that are crucial in the sterile manufacture of pharmaceutical products. The OOS results are due to possible risk presence in the manufacturing company of sterile pharmaceutical products.
Where the risk itself can be defined as "An event in the manufacture, control and supply of a drug that potentially has an adverse health effect". However, in the case of OOS of Grade D filing rooms, the risk comes mainly from extrinsic factors that include environment, personnel and surfaces. Of all three of these, staff presence and flow staff are considered to be the most important source of contamination in the manufacturing environment (BioPhorum, 2019).
CONCLUSION AND RECOMMENDATION 4.1. Conclusion
Recommendation
SELF REFLECTION
Marrë nga https://www.fda.gov/inspections-compliance-enforcement-and-criminal-investigations/inspe ction-guides/pharmaceutical-quality-control-labs-793.
