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9040 WASHING AND STERILIZATION*

Sterilize all contaminated laboratory ware before cleaning to times are needed to kill spores and very resistant microorgan-
prevent potential contamination to personnel handling contami- isms. Sterilize glassware in metal containers at 170°C for not less
nated material. Remove all markings before initiating washing than 2 h. For all bottles, loosen caps before autoclaving. If
sequence. If material has dried on glassware, a pre-soaking may desired, moisture present in empty sterile containers after auto-
be needed. If mechanical laboratory washers are used, equip claving may be removed by placing items in a drying oven at
them with influent plumbing of stainless steel or other nontoxic 100°C for 10 to 15 min.
material. Do not use units designed for home use. Ensure that the As long as the bromthymol blue test is being done on each
spray from water jets reaches all parts of deep vessels. Do not batch of washed glassware, run the toxicity test, ¶ 9020B.5a2),
use copper plumbing to distribute water. Use stainless steel or before initial use of a washing compound and whenever a new
other nontoxic material for the rinse-water system. Consistently formulation or washing procedure is used. If the bromthymol
follow the same washing procedures whether using an automated blue test is not done consistently, run the toxicity test on a per-lot
system or washing by hand. or annual basis, whichever is more frequent. Review Section
Cleanse all glassware and plasticware thoroughly with a suit- 9020B for quality control checks on autoclaves and sterilized
able detergent and warm water. To remove all traces of residual bottles.
washing compound, rinse five to ten times with cold water after
bubbles/foam are gone. In addition, rinse two to three times with Bibliography
reagent-grade water.
If desired, dry glassware before use by placing in a drying BORDNER, R.H., J.A. WINTER & P.V. SCARPINO, eds. 1978. Microbiolog-
oven at 100°C for 10 to 15 min. ical Methods for Monitoring the Environment, Water and Wastes.
Glassware may be sterilized, except when in metal containers, EPA-600/8-78-017, U.S. Environmental Protection Agency, Envi-
by dry heat for 2 to 4 h at a temperature of 170°C. Longer times ronmental Monitoring & Support Lab., Cincinnati, Ohio.
are needed to kill cryptosporidia and denature prions. Alterna- AMERICAN SOCIETY FOR TESTING AND MATERIALS. 1998. Standard Practices
for Cleaning Laboratory Glassware, Plasticware, and Equipment
tively, add a small amount of distilled water (to prevent airlock)
Used in Microbiological Analyses. D5245-92 (reapproved 1998),
and autoclave at 121°C for at least 30 min. Longer sterilization American. Soc. Testing & Materials, West Conshohocken, Pa.
U.S. ENVIRONMENTAL PROTECTION AGENCY, 2005. Manual for the Certi-
* Approved by Standard Methods Committee, 2006.
fication of Laboratories Analyzing Drinking Water, 5th ed. EPA
Joint Task Group: Margo E. Hunt (chair), Ellen B. Braun-Howland, Terry C. 805-R-05-004, U.S. Environmental Protection Agency, Cincinnati,
Covert, Gil Dichter, Nancy Hall, Robin Oshiro. Ohio.