Endotoxin test for

See also U.S. Department of Health s Guideline on Validation of the LAL Test as an End-Product Endotoxin Test for Human and Animal Parenteral Dmgs, Biological Products and Medical Devices. ... 

LAL) test. Additionally the agency has approved the use of the bacterial endotoxins test for many drug and device products. 

The quality control of the final product must be carried out before release of the batch (except for the sterility and the endotoxin tests for extremely short-lived radionuclides). Consequently, all procedures must not only be very fast but also very accurate, and in all cases it is very important to have a properly established quality assurance system that might permit parametric release of the produced batches. The quality control assays that must be carried out in the radiopharmaceutical includ the following ... 

The first large-scale conversion of the rabbit test to the BET occurred in Supplement 5 to USP XXII when the BET became the official endotoxin test for 185 USP articles. The USP 24 has endotoxin limits for over 650 USP articles. 

The results of endotoxin tests for in-process solutions, bulk materials, and finished parenteral products should be reported in the same units as those assigned to the product. Two factors determine the sensitivity of a BET. For infusion solutions and device extracts, the gel-clot sensitivity or the lowest point on the standard curve (lambda for kinetic LAL) and the amount of dilution determine test sensitivity.For products that have an endotoxin limit in EU/mg, the choice of lambda and the concentration of the test material determine sensitivity. The formula for product-specific sensitivity (PSS) is a convenient way to calculate the sensitivity of a BET for this type of product, where ... 

Guideline on validation of the Limulus Amebo-cyte test as an end-product endotoxin test for human and animal parenteral drugs, biological products and medical devices. Food and Drug Administration Rockville MD, 1987. 

Standardization and Testing". The final vaccine is tested for safety, potency, and residual chemicals. Safety includes testing for endotoxin and stetihty. Potency is evaluated by quantitative determination of the amount of hemagglutinin in the vaccine. Antibody to this glycoprotein is associated with protection. The single radial immunodiffusion (SKID) technique is used to standardi2e the mass of this protein in comparison to a reference preparation. 

The antagonistic properties of FR 900452 have also been examined in a model of endotoxin shock, the haemodynamic and haematological manifestations of this condition closely resembling the changes induced by PAF. FR 900452 (10 mg/kg, i.v.) almost completely prevents PAF (1 /ig/kg, i.v.)-induced thrombocytopenia and leukocytopenia in rabbits [295]. It also significantly inhibits endotoxin (E. coli LPS, 30 /ig/kg, i.v.)-induced thrombocytopenia but not leukocytopenia. The same dose of FR 900452 also causes the decreased arterial blood pressure to return to normal in the endotoxin-induced rat hypotension model, an effect also reported for other PAF inhibitors [121, 274], Finally, FR 900452 has been tested for its therapeutic effect on rat nephrosis induced by aminonucleoside (puromycin, 100 mg/kg, i.p.). At 100 mg/kg twice a day orally for 6 days, the agent significantly reduces urinary protein loss in nephrotic rats [298]. 

The opposite of harmony is disharmony. An example of disharmony is the need to repeat tests using rabbits for pyrogen where testing for bacterial endotoxin is otherwise prescribed. This represents the most extreme disharmony of methods. But there is even a greater disharmony, and that would be to reach different eonelusions as to pass/fail the speeimen. In this ease, the quality control professionals must make a judgment as to whether or not this material can be sold in one or more regions. Functionally equivalent to harmonization is the absence of disharmony. Because of a difference in policy, pharmacopoeias may not differ on adoption of a test at all. If certain tests are eonsidered neeessary by one pharmacopeia in order to protect the consumer, it is appropriate for that pharmacopeia to adopt the test without reference to any other region. 

These tests would include compendial tests for Microbial Limits, Sterility, Bacterial Endotoxins, and Antimicrobial Effectiveness. 

Injectables, Inhalation solutions a. Sterility test, b. CCI, AET. c. Bacterial endotoxin test Test all batches on stability with the exception of the first three batches for AE a. b. c. 0 only 0, 12, 24, 36 months 0 and expiry only... 

All injectable products should be tested for endotoxin at release. 

The main objective of validation of an analytical procedure is to demonstrate that the procedure is suitable for its intended purpose. The procedures presented in this SOP provide basic guidelines for the validation of methods for microbiological assay, estimation of the number of microorganisms, detection of indicators of objectionable microorganisms, validation of preservative efficacy testing, and validation of the sterility testing and endotoxin test (LAL test). 

Product component bioburden. Endotoxin analysis involves testing twenty (20) pieces. Twenty (20) vials or stoppers are filled with or immersed in WEI, mixed and sonicated. Duplicate 0.1 mL aliquots are removed and tested for endotoxin per the USP <85> Bacterial Endotoxins Test. 

If the hll volume of an end product is less than 5 ml, a minimum of ten samples will be taken for endotoxin testing. 

Two independent replicate solutions of the preparation being examined at the dilution with which the test for interfering factors was completed and endotoxin standard BRP at a concentration of 2X. 

Excipients used in injectable formulations have to meet several stringent requirements. A positive identification test uniquely applicable to the excipients is required (e.g., infrared spectrophotometry and chromatography). It is important that manufacturers identify and set appropriate limits for impurities. These limits should be based upon appropriate toxicological data, or the limits described in national compendial requirements. Manufacturing processes should be adequately controlled so that the impurities do not exceed such established specifications. Solvents or catalysts used in the excipient production process should be removed to appropriate levels. If naturally derived, excipients should meet endotoxin levels and may require further testing for bovine spongiform encephalopathy (BSE) /... 

Failure to establish yields or acceptable levels of rejects for both in-process and finished product Failure to conduct stability studies Manufacturing equipment not identified and/or qualified Inadequate training of employees working in aseptic operations Inadequate process change procedures Validation protocols that lack acceptance criteria Incomplete investigations of laboratory failures Failure to follow United States Pharmacopeia (USP) procedures for the bacterial endotoxin test... 

The in vivo assays for the Cox-2 inhibitors are essentially those used historically for NSAIDs to evaluate both their desired effects on inflammation, pain, and fever and their undesired effects, mainly on GI lesions. The primary in vivo assay for anti-inflammatory efficacy is the carrageenan-induced rat paw edema assay, and 51 Cr fecal excretion is used to test for damage to the intestinal mucosa in rats and in squirrel monkeys. Additional tests for efficacy are the endotoxin-induced pyresis in rats and squirrel monkeys for control of fever and the carrageenan-induced rat paw hyperalgesia assay for analgesic efficacy. These in vivo assays have been described (Chan et al., 1995 Chan et al., 1997). The rat... 

Based on rFC, a modern and yet simple, rapid, specific and sensitive diagnostic test for endotoxin has been developed (Ding and Ho, 2001). rFC is a proenzyme until it encounters trace levels of endotoxin where it unequivocally exhibits full enzymatic activity, hence, acting as a very sensitive and specific biosensor for endotoxin. The resulting activated rFC acts as a catalyst to hydrolyse a synthetic substrate to form a quantifiable product, which measures the level of endotoxin. A fhiorimetric assay... 

Inflammation is a common component associated with sepsis, meningitis, as well as respiratory tract, urinary tract, viral, and bacterial infections (Table 1). Bik is elevated during bacterial or viral infection. The presence of urinary Bik correlates well with standard urinalysis tests for urinary tract infections [20]. Endotoxins released from infectious pathogens induce inflammation and immune cell activation. Macrophages release interleukins and cytokines (IL-1, IL-6, IL-12, IL-15, IL-18, TNF-a) on exposure to lipo-polysaccharide (LPS) and lipoteichoic acid (LTA) endotoxins. These cytokines act as a chemotactic factors causing immune cell migration to the site of the infection followed by activation and release of proteases. Cytokines also induce increased vascular permeability in the endothelial. Bik suppresses further cytokine release by protease and intern additional migration and activation of immune cells. Additionally, a stabilization of the immune cell membrane prevents further release of proteases [4]. 

Bioburden testing is required for biologies used in pivotal preclinical studies to support clinical development and registration. Endotoxin levels are the primary concern in this regard. There are several major regulatory documents listed below (taken from Associates of Cape Cod Inc. Web site http //www. acciusa.com/) that describe how drugs, devices, dialysate, water, and other substances are to be tested for endotoxin. 

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