The bacterial endotoxin test (BET) is performed in most modern laboratories under controlled conditions as the important factor to avoid interference.
Appropriate aseptic technique is important when preparing and diluting standards and handling samples. Gowning practice outside of normal laboratory personal protective equipment (PPE) requirements is not a concern unless the product under test demands specific analyst safety considerations due to toxicity or infectiousness. Gloves should be TALC-free, as the TALC may contain significant levels of endotoxins. Plate readers, water baths, and dry heat blocks used for sample incubation should be on a laboratory bench away from heating, ventilation, and air conditioning (HVAC) ducts, significant vibration, and laboratory traffic that could affect the test results. Sample hold times and conditions should be determined and subsequently documented, if necessary, to ensure that accurate test results can be generated in the qualified time.
For example, if the laboratory receives a Water for Injection (WFI) or in-process sample, must it be refrigerated or can it remain at room temperature, and for how long? Prior to testing, it is recommended that the primary sample container(s) be adequately mixed before removing the test aliquot(s) for either direct testing or subsequent dilution.
Bioendo Bacterial Endotoxin Test, the experiments are including gel clot method endotoxin test assay and quantitative endotoxin test assay, gel clot method endotoxin test assay is qualitative endotoxin detection, these experiment requires the consumables is depyrogenation processing, such as endotoxin free reaction tubes, dilution tubes and pyrogen free tips; quantitative endotoxin detection have kinetic chromogenic endotoxin test, kinetic turbidimetric endotoxin test, these experiment requires consumables have to meet the top level of endotoxins less than 0.005EU/ml ( 0.001EU/ml ), such as endotoxin free tubes, pyrogen free tips, and pyrogen free microplates, even pyrogen free resevoir. By the way, if the samples treatment, the container have to be the endotoxin free sample bottle.
In endotoxin testing, interference can arise from a variety of sources, such as sample matrix components, test reagents, or equipment.
To avoid experiment interference, the following measures can be taken:
1. Sample Preparation: Proper sample preparation is essential for accurate endotoxin testing.
The sample matrix should be thoroughly tested and optimized to ensure compatibility with the endotoxin assay.
In particular, interfering substances such as lipids and proteins should be removed or minimized using appropriate techniques such as filtration or centrifugation.
2. Positive and Negative Controls: It is essential to include positive and negative controls in the assay to monitor for interference.
Positive controls confirm the functionality of the test, while negative controls detect any contamination or interference from the assay components.
3. Quality Control: Quality control should be performed on all reagents, equipment, and water used in the assay.
This ensures that the reagents are free from endotoxin contamination and are working correctly.
4. Standardization: The assay should be standardized to ensure that all results are comparable and reproducible.
This involves the use of a standard curve to calibrate the assay and the use of standardized techniques for sample preparation, incubation, and detection.
5. Validation: The assay should be validated to ensure that it is specific, sensitive, and reliable.
This involves testing a range of samples, including those known to contain endotoxin, to determine the accuracy and precision of the assay.
By following these measures, interference can be minimized, and accurate endotoxin testing can be achieved.
Post time: Dec-01-2022
