Sterility testing administration

FDA draft guidance for industry. In Container and Closure Integrity Testing in Lieu of Sterility Testing as a Component of the Stability Protocol for Sterile Products, Food and Drug Administration Rockville, MD, 1998. 

The contaminated bottles were not detected by end-product sterility testing. The batch was released to a wholesaler and distributed to the Devonport Section of Plymouth General Hospital in March 1972. The high concentrations of microorganisms found in the infusion fluid can be attributed to the period of time betweeit sterilization, distribution, and final administration to the patients. 

Radiopharmaceuticals for parenteral administration should be sterile. Preparation under aseptic conditions should ensure this however, conditions and operator dispensing technique can be checked by carrying out sterility testing of products. 

Recognizing these problems, UK food regulatory authorities have generally abandoned the use of quantitative microbial counts as enforceable standards of food quality. Despite this, the European Pharmacopoeia has introduced both quantitative and qualitative mierobial standards for non-sterile medicines, which might become enforceable in some member states. It prescribes varying maximum total microbial levels and exclusions of particular species according the routes of administration. The British Pharmacopoeia has now ineluded these tests, but suggest they should be used... 

If it is assumed that the radiation sterilizer equipment and facilities have been qualified and microbiological studies have been conducted as previously outlined, the next step in the validation process is the complete evaluation of the radiation sterilization cycle. Tests are conducted to determine the effect of minimum and maximum product density on the ability of the minimum or nominal radiation dose—determined during the microbiological studies to produce a given log reduction in the biological indicator population—to sterilize the load. For example, it was found that a 0.2-Mrad dose of cobalt-60 will produce a 1-log reduction in the population of B. pumilus. The microbial load of a one-package polyvinyl chloride (PVC) device (intravenous administration site) was estimated to be approximately 1000. A probability of a nonsterility level of 10 6 is desired, therefore theoretically, the minimum dose necessary to produce a 9-log reduction in the microbial population is 1.8 Mrad. 

Method The principle of GEC involves the intravenous administration of galactose in excess substrate in order to assess the maximum rate of hepatic metabolization. This requires a galactose concentration of 0.5 g/kg BW (serum concentration of > 45 mg/dl for at least 45 minutes), which is injected within 4—5 minutes as a 40% pyrogen-free sterile galactose solution. The bladder has to be emptied immediately prior to the injection and subsequently the urine has to be collected for exactly 5 hours to assess the respective galactose excretion (in grams). Blood samples are taken from the fingertip (or from the cubital vein into an EDTA test tube) at 5, 25 and 45 minutes after injection. We have reported in detail on how to conduct this method as well as on the analysis of GEC. (67, 69)... 

The pharmacopeial standard applying to sterile products is that they must be capable of passing a Test for Sterility. A Test for Sterility is described in U.S. Pharmacopeia (USP) under Section 71 and in the European Pharmacopoeia (PhEur) under Section 2.6.1. These were harmonized along with the Japanese Pharmacopoeia and the requirements of the Australian Therapeutic Goods Administration in 1999, but they still have some minor differences in detail. 

Parenteral dosage forms require extensive testing of sterility and biological impurity contamination because of the high sensitivity of this administration route. These same sterility standards do not apply to solid and... 

To determine the nociceptive threshold of the mice, the hot plate test and the tail flick test were used. Drugs used are dissoved in sterile distilled water from icv administration immediately prior to injection at 5 pl/kg or 5pi mouse, respectively. The icv injection was performed and the site of administration of the peptide was verified in all animals by the injection of 1% methylene blue and the examination of the dye distribution in the cerebral ventricles at the termination of the experiment. To evaluate the hot plate and tail flick test responses detailed below, a control latency (To) was obtained from the mean of two latencies determined prior to drug injection test latencies (T1) were determined at various times after injection for each animal. The percentage of analgesia was calculated as (Ti -To)/ (T2 -To) x 100 where the cut off times (T2) for the hot plate and tail flick test were 60 and 15s, respectively. The median antinociceptive dose (ED50) and 95% confidence limits were calculated according to the method of Litchfield and Wilcox. 

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