Endotoxin content

Cotton dust activates complement vitro by both the classical (antibody dependent) and alternative (antibody independent) pathways (12,47,48). It is proposed that endotoxins may be the agents responsible for complement activation (49). Cotton dust extractions maximizing endotoxin content are 10 times more potent than other extracts in activating complement (12). Activation of complement via the alternative pathway has also been... 

Histamine is released from whole rabbit blood by both cotton extracts and endotoxins, however, the ability of AECD to release histamine is greater than would be expected from endotoxin content alone. Antweiler (63), however, was unable to show in vivo histamine release by endotoxins obtained from cotton extracts in rabbits nor could he show an acute fall in blood pressure of cats after I.V. endotoxin injection, as occurred with subsequent injections of compound 48/80 or other dust extracts. He concludes that endotoxins are not responsible for any histamine releasing activity of cotton dust. 

Cinkotal et al. (11) measured the endotoxin content In the cardroom of seven cotton mills, and two cotton waste mills. None was found in a tea packing factory or a pipe tobacco factory and the level was low in the willowing mills. Bysslnotic symptoms correlated with the level. 

Fischer et al. (21) demonstrated that the endotoxin content of raw cotton is significant (300 ng/gm lint) and is markedly decreased by washing and/or bleaching of the cotton, and by heating the cotton in hydrogen peroxide. Only prolonged high heat results in a complete loss (5 log decrease) of endotoxin activity from cotton dust. 

Prokaryotic expression E. coli Rapid growth with high yields (up to 50% total cell protein) Extensive range of vectors Simple, well-defined growth media Lack of post-translational processing Product may prove toxic to host Product incorrectly folded and inactive High endotoxin content... 

With respect to endotoxin content, the FDA requires that each dose have less than 0.5 endotoxin units (EU) for each ml of... 

Belanich et al.63 reported the removal of endotoxins from protein mixtures. Endotoxins in a bacterial extract containing a protein photolyase was passed through a stack of 10 disk membranes (Q-type, Sartorius). LAL assay was used to monitor the endotoxin levels after each pass. There was over 5 log reduction in endotoxin content after three passes through the membranes. The protein content was reduced during this process, however, the enzyme s specific activity was increased 35-fold. This study also determined that the binding capacity of the membrane was greater than 2.25 million EU/cm2 of membrane area. 

The BFS and FFS are unique systems in that the final container is formed as a sterile container just prior to the aseptic filling step. The BFS requires careful control over the endotoxin content of the LDPE (and other polymeric materials) beads used to create the containers as well as the melting conditions utilized to form them. 

Endotoxin contamination has been encountered frequently in Hb solutions. Historically, the production of Hb solutions low in endotoxin content has proven challenging, in part because much of the early work was performed in academic research laboratories that were not familiar with the techniques used in the pharmaceutical industry to prevent bacterial contamination of the process stream. In addition, Hb has been reported to bind endotoxin, making it even more difficult to completely remove once present. Therefore, the production of Hb solutions must be carefully designed to minimize the introduction of endotoxin into the process stream. 

Baraff LJ, Manclark CR, Cherry JD, Christenson P, Marcy SM. Analyses of adverse reactions to diphtheria and tetanus toxoids and pertussis vaccine by vaccine lot, endotoxin content, pertussis vaccine potency and percentage of mouse weight gain. Pediatr Infect Dis J 1989 8(8) 502-7. 

Pyrogenicity or endotoxin content. The pyro-genicity of a specified dose of product when administered to rabbits can be assayed by a standard pharmacopoeial method but the trend is to replace this with an in vitro assay for endotoxin (Chapter 19). The capacity of the product to induce gelation of Limulus polypbemus amoebocyte lysate is determined against a reference endotoxin preparation and the result is expressed as IU of endotoxin. 

Operate the system per applicable SOPs. Perform sampling over a 1-month period per the sampling procedure and schedule. Test samples for conformance to current USP Water for Injection monograph, microbial content, and endotoxin content. Identify all morphologically distinct colony forming units (CFUs) to at least the genus level. 

The endotoxin content of all endotoxin indicators must be reduced to levels of <0.025 EU (or undetectable), or at least demonstrate a minimum 3 log reduction for three (3) consecutive runs. 

Another kind of biotechnological product that is acquiring interest as a therapeutic agent is the biopharmaceuticals derived from plants. As a result of the limited production capacity of many pharmaceutical products, the production in plants represents an innovative tool that can expand the yields of active principles because the large volume of biomass that can be developed at the fields. An extra advantage is that these products are free of human diseases and mammalian viral vectors. In products derived from bacteria, the bacterial endotoxin content of the... 

Furrer et al. have tested different solvents to recover PHA and tested the influence on the toxicity of the polymer. The results showed significant differences in endotoxicity of PHA due to the use of different solvents. Ethyl acetate, acetone, tetrahydrofuran (THF), 2-propanol and t-hutyl methyl ether (MTBE) extracts showed a much lower endotoxin content than the extracts with methylene chloride (CH2CI2) and hexane. ... 

The use of oxidizing agents such as hydrogen peroxide or henzoyl peroxide has also reduced the endotoxin content to less than 20 endotoxin units/gram of PHA, and this PHA did not result in an acute inflammatory response when implanted in animal models. ... 

The Ph. Eur. gives limits for radiopharmaceuticals in general but for some individual radiopharmaceutical preparations as well. In most radiopharmacy departments the endotoxin content of radiopharmaceutical preparations is not tested before injection. In some situations, e.g. development of a new preparation process or when using generators for longer periods of time (weeks to some months) endotoxin testing may be useful (see further Sect. 19.3.4). 

Water for injections in bulk must comply with the requirements as formulated for Purified water. It must also be produced free of bacterial endotoxins and stored such that it remains free of them. It can be used for parenteral products and irrigations that are terminally sterilised. Water for injections, sterilised, must meet the requirements of sterility (sterility test) and the bacterial endotoxin content must not exceed 0.25 lU per mL. This is used for parenteral products and irrigations that are not terminally sterilised and for dissolving powders for injection immediately before use. 

A report by Karol et al. (1985) showed that the potency of cotton dusts from three production areas as measured by the respiratory behavior of guinea pigs was positively correlated with their endotoxin contents. 

Airborne dusts were collected during the carding operations with vertical elutriators which operated in the exposure room which was remote the card room (Cocke and Bargeron, 1984). Thirty-seven millimeter, 5ym pore size filters with collected dusts were provided to our laboratories for analysis of endotoxin content. The filters represent dust which was collected throughout the exposures, and all samples were labeled by number. No reference to the variety or area of growth was made until after the endotoxin analyses were completed. 

Figure 1. Mean Endotoxin Contents in Airborne Dusts Generated During the Carding of 3 Varieties of Cottons Grown in California During Crop Years 1982 and 1983.

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