The pyrogens often encountered in medicines are bacterial endotoxins. In the quality control of manufactured medicines, one can accept the view that the absence of endotoxin means the absence of pyrogens. Compared with the traditional pyrogen inspection method, the home-free pyrogen inspection method, the limulus reagent method has the advantages of high specificity, high sensitivity, quickness, and simplicity in detecting bacterial endotoxins. However, because there are interference factors in blood products, they have inhibitory or strengthening effects that affect the testing of endotoxins. Therefore, this interference effect needs to be ruled out.
Fig1. Protein structure of human albumin.
10% human serum albumin is diluted 3 times or more without inhibition or enhancement and does not interfere with bacterial endotoxin test. Therefore, using a specific limulus reagent with a sensitivity of 0.125 EU/ml, human serum albumin is diluted 3 times bacterial endotoxin test.
Creative BioMart uses Limulus Amebocyte Lysate test for human albumin endotoxin testing. The primary application for LAL is the testing of blood or cerebrospinal fluid. There are three basic methodologies: gel-clot, turbidimetric, and chromogenic. The primary application for LAL is the testing of parenteral pharmaceuticals and medical devices that contact blood or cerebrospinal fluid. In the United States, the FDA has published a guideline for validation of the LAL test as an endotoxin test for such products. Limulus amebocyte lysate (LAL) is an aqueous extract of blood cells (amoebocytes) from the Atlantic horseshoe crab, Limulus polyphemus. LAL reacts with bacterial endotoxin lipopolysaccharide (LPS), which is a membrane component of gram-negative bacteria. This reaction is the basis of the LAL test, which is widely used for the testing and quantification of bacterial endotoxins.
Fig2. Workflow of limulus Amebocyte Lysate test.
Pyrogen testing is one of the important inspection items to ensure the safety of drugs. In 1923, Seibert first proposed the method of detecting pyrogens in rabbits. In 1942, the United States Pharmacopoeia first used the rabbit method as a pyrogen test for drugs, which played an important role in ensuring drug quality and drug safety. Rabbit pyrogen test can objectively reflect the characteristics of pyrogens in various drugs and biological products in the body, and can intuitively reflect the temperature rise of experimental rabbits caused by pyrogens in drugs.
Because the test results of the two methods, the home immunity method and the gadolinium reagent, are relatively high, most of the samples were qualified by the reward method, and no samples that failed the rabbit test and failed the pot test were found. Therefore, in the quality control of semi-finished products, it is of great practical significance to use the combination of the pot method and the home immunity method for endotoxin inspection. In addition, the limulus reagent method is also beneficial to evaluate the endotoxin content level of the product. For human albumin samples that are positive for the provincial test result after a 3-fold dilution, the dilution factor is increased until the dilution factor when the test result is negative, which is called the maximum dilution factor.
Creative BioMart offers a corresponding endotoxin testing service. You can purchase the corresponding endotoxin removal kit and related accessory products according to the needs of your own samples. We guarantee that all instruments, water, reagents, and consumables used in the experiment are free of endotoxins, and the experiment is conducted in a clean room to ensure that low levels of endotoxins are returned to the sample. In addition, we can also provide you with related services including:
|Project name||Endotoxin testing in human albumin|
|Testing purpose||The pyrogens often encountered by human serum albumin are bacterial endotoxins. In the quality control of finished products, endotoxins must be detected.|
|Testing cycle||3-5 days.|
|Service including||We provide you with raw data and test reports.|