Pyrogen-free

Biosynthesis ofS(— )-M llc Acid. Aqueous fumaric acid is converted to levorotatory malic acid by the intracellular enzyme, fumarase, which is produced by various microorganisms. A Japanese process for continuous commercial production of S(—)-mahc acid from fumaric acid is based on the use of immobilized Brevibacteriumflavum cells in carrageenan (32). The yield of pyrogen-free S(—)-mahc acid that is suitable for pharmaceutical use is ca 70% of the theoretical. 

Bacteriostatic water for injection is sterile and pyrogen-free and contains bacteriostatic agents. The dmg involved must be compatible with the antimicrobial agents present. 

In Parenteral and Enteral Nutrition. Amino acid transfusion has been widely used since early times to maintain basic nitrogen metaboHsm when proteinaceous food caimot be eaten. It was very difficult to prepare a pyrogen-free transfusion from protein hydrolysates. Since the advances in L-amino acid production, the crystalline L-amino acids have been used and the problem of pyrogen in transfusion has been solved. The formulation of amino acid transfusion has been extensively investigated, and a solution or mixture in which the ratio between essential and nonessential amino acid is 1 1, has been widespread clinically. Special amino acid mixtures (eg, branched chain amino acids-enriched solution) have been developed for the treatment of several diseases (93). 

The ability to remove particulates has made RO indispensable in the production of ultra-pure water for microchip washing. Its ability to remove large molecules enables it to produce pyrogen-free water for the pharmaceuticals industry. In the USA and elsewhere RO is permitted for producing the water used in making up injectable preparations. The European Pharmacopoeia still insists on distillation for this, but the larger amounts of water needed for ampoule washing, etc. are often purified by RO. 

Parenteral dose forms include aqueous, aqueous organic, and oily solutions, emulsions, suspensions, and solid forms for implantation. These parenterals need to be sterile and pyrogen-free they are, if possible, buffered close to normal physiological pH and preferably are isotonic with the body fluids. 

Sterile producfs for injection represent a particular challenge for the pharmaceutics development group. To prepare injectables, the pharmacists need not only sterile rooms in which to work at the laboratory, pilot plant, and production scales of operation, but they also require pyrogen-free wafer. Pyrogens are impurities, generally originating with... 

After weighing the dust filters, the amount of endotoxin was determined by shaking them In 10 ml of pyrogen free water and preparing serial dilutions. Llmulus lysate (Cape Cod Associates Inc.) was added to the dilutions according to the manufacturer s recommendations. The last dilution giving a stable clot was read as the Escherichia coll endotoxin equivalent concentration. Dilutions were also prepared with commercial E. coll endotoxin (. coll 026-B26, Dlfco) to assess the accuracy of the production reference standard. The values were always found to agree closely with the stated values. 

Dextran, pyrogen-free average mol wt 100,000-200,000 Dalton (United States Biochemical Corp., Cleveland, OH). 

Finally, radiopharmaceuticals are often prepared on a daily basis within the framework of clinical studies which often last several months or years. They demand a viable and reproducible production chain, leading to a sterile- and pyrogen-free radiopharmaceutical of high radiochemical purity. Therefore, microprocessor-controlled automated synthesis devices [31] are developed in order to ensure routine pharmaceutical production. They are becoming mandatory in order to meet the demands related to Good Laboratory Practice (GLP) and Good Manufacturing Practice (GMP). 

Bacterial enck)toxin pass the water through a pyrogenic connection for about 1 minute and then collect the sample in a pyrogenic bottle. All materials coming in contact with test materials and reagents must be pyrogen free and careful technique is essential to prevent contamination with environmental endotoxin. 

Using a sterile, nonpyrogenic pipette tip and pipettor, aseptically pipette 5.0 ml of pyrogen-free water into the endotoxin vial. 

Gently mix the stoppers in the container to ensure that all surfaces come in contact with the water and sonicate for 30 minutes. Return the container to the laminar flow hood. Open the container and, using an automatic pipettor fitted with sterile, nonpyrogenic tips, pipette 0.1 ml of the water into each of two sterile pyrogen-free test tubes. 

Positive controls pipette duplicate 0.1-ml aliquots of freshly prepared endotoxin standard solution, bracketing the labeled lysate sensitivity, into separate 10 X 75 mm pyrogen-free test tubes. 

Negative controls pipette 0.1-ml sterile pyrogen-free water into two 10 X 75 mm test tubes. 

Working under a laminar flow hood, add sterile pyrogen-free WFI to the vials to be tested. The following amounts are added ... 

Pool water from all 20 vials into one sterile pyrogen-free container. Vortex to mix. Pipette two aliquots of 0.1 ml each into labeled 10 X 75 mm sterile pyrogen-free glass tubes. 

Pipette duplicate 0.1-ml aliquots from each vial into labeled 10 x 75 mm sterile pyrogen-free glass tubes. Follow steps 10 through 14 in the stopper section of the SOP. Record results. 

A sample (100 ml) of prefiltered solution will be taken in a sterile pyrogen-free container compounding. 

Using aseptic technique, open the required number of vials, vor-texing each prior to removing an aliquot. Pool the samples aseptically in a sterile, pyrogen-free vial. Eor in-process sample, treat the bulk solution as a pooled sample. 

If the hll volume of a product is 10 ml or more, take an equal volume of the product aseptically from each of the vials to be tested and pool these samples in a sterile, pyrogen-free vial. 

Use this concentrate for making appreciate serial dilutions. Use 13 X 100 mm pyrogen-free culture tubes. 

Take two 10 x 75 mm sterile, pyrogen-free glass test tubes. 

Sterile forceps Vacuum pump Laminar flow hood Water bath set at 45 to 55°C Incubator set at 30 to 35°C Mechanical shaker Sterile surgical gloves Self-contained anaerobe jar Bunsen burner Sterile, pyrogen-free WET... 

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