Pyrogenicity

The General Tests and Assays. This section of the USP gives methods for tests that are general in nature and apply to a number of the substances. Procedures are iacluded for such tests as heavy metals, melting point, chloride, sulfate, sterility, bacterial endotoxins, and pyrogens. Also iacluded are descriptions of various analytical techniques, such as spectrophotometry, chromatography, and nmr, and descriptions of tests to be used on glass or plastic containers, mbber closures, etc. 

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. 

Pharmaceutical Industry. In the pharmaceutical industry, sterility of deionized water systems is maintained by using an ozone residual. The ozone residual concentration is maintained at >0.3 ppm ppm in the water recirculation loop. Prior to product compounding, the ozone residual is removed by contact with uvirradiaton for <1 s. Ozone also is used to oxidize pyrogens from distilled water destined for intravenous solutions. 

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). 

Amorphous sihca exists also ia a variety of forms that are composed of small particles, possibly aggregated. Commonly encountered products iaclude sihca sols, sihca gels, precipitated sihca, and pyrogenic sihca (9,73). These products differ ia their modes of manufacture and the way ia which the primary particles aggregate (Fig. 8). Amorphous sihcas are characterhed by small ultimate particle si2e and high specific surface area. Their surfaces may be substantially anhydrous or may contain silanol, —SiOH, groups. These sihcas are frequentiy viewed as condensation polymers of sihcic acid, Si(OH)4. 

Fig. 8. Different forms of commercially important amorphous silica (a) sol, (b) gel, (c) precipitate, and (d) pyrogenic.

The diversity of sihca forms and their properties leads to a broad range of apphcations. Reviews on usage iaclude those on crystalline and vitreous sihcas (94), on sihca sols (95), on sihca gels and precipitated sihcas (96), and on pyrogenic sihcas, precipitated sihcas, and sihca gels (97). 

Fig. 6. Transmission electron micrograph of a commercial pyrogenic siUca, Wacker HDK. Magnification of 225, OOOx. ...

The typical range of properties for commercial coUoidal silicas, silica gels, precipitated silicas, and pyrogenic silicas is given in Table 1. 

Property CoUoidaln sUica SUica gels Precipitated sUica Pyrogenic sUica... 

Fig. 15. Schematic of pyrogenic siUca production (54). See Figure 13 for definitions.

Reaction with Hydrogen and Oxygen. The combustion of sihcon tetrachloride in a hydrogen oxygen (air) flame is an important technology leading to fumed or pyrogenic siUca. 

Ultra filtration. This process removes macromolecules, microorganisms, particulate matter, and pyrogens using a thin, selectively permeable membrane. Ultrafiltration caimot remove ions from water and is generally employed as a polishing process. 

Factors to be considered in maldng the selection of chromatography processing steps are cost, sample volume, protein concentration and sample viscosity, degree of purity of protein product, presence of nucleic acids, pyrogens, and proteolytic enzymes. Ease with which different types of adsorbents can be washed free from adsorbed contaminants and denatured proteins must also be considered. 

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