Pennsylvania Code › Title 25 - ENVIRONMENTAL PROTECTION › Part I - Department of Environmental Protection › Subpart D - Environmental Health and Safety › Article VI - General Health and Safety › Chapter 252 - ENVIRONMENTAL LABORATORY ACCREDITATION › Subchapter D - QUALITY ASSURANCE AND QUALITY CONTROL REQUIREMENTS › Section 252.404 - Essential quality control requirement- microbiology

Current through Register Vol. 54, No. 28, July 13, 2024
(a) In addition to the requirements of §
												252.401 (relating to basic
						requirements), environmental laboratories performing testing or analysis in the
						area of microbiology shall comply with this section.
(b) When the method selected by an
						environmental laboratory in accordance with §
												252.307 (relating to methodology)
						contains more stringent requirements than the requirements of this section, the
						environmental laboratory shall follow the more stringent requirements contained
						in the method.
(c) The following
						pieces of equipment shall be maintained according to this subsection:
(1)
Autoclave.
(i) An environmental laboratory shall use
						autoclaves that meet specified temperature tolerances of the method. Pressure
						cookers may not be used.
(ii) A
						continuous temperature-recording device or a maximum-temperature-registering
						thermometer shall be used during each autoclave cycle.
(iii) An environmental laboratory shall
						verify the sterilization capability of each autoclave by utilizing appropriate
						biological indicators (for example, spore strips or ampoules) once a month.
						Records of biological indicator tests shall be maintained in a laboratory
						notebook and include the autoclave identification, date, incubation time and
						temperature, results and initials of the responsible individual.
(iv) An environmental laboratory shall verify
						the mechanical timing device, if used, for each autoclave every 3 months.
						Records of mechanical timer verification shall be maintained in a laboratory
						notebook and include the autoclave identification, date, mechanical timing
						device time, actual time and initials of the responsible individual. Correction
						factors shall be documented and used.
(v) Autoclaves shall be properly cleaned and
						maintained. Copies of service contracts or internal maintenance protocols and
						maintenance records shall be kept.
(vi) Required times for autoclaving items at
						121°C are set forth in this subparagraph. The following items must be at
						temperature for the required amount of time. Except for membrane filters and
						pads and carbohydrate-containing media, indicated times are minimum times and
						may necessitate adjustment depending upon volumes, containers and loads. For
						autoclave runs that include membrane filters and pads and media, the total
						cycle time may not exceed 45 minutes. Autoclaved membrane filters and pads and
						media shall be removed immediately after completion of the autoclave cycle.
(A) Membrane filters and pads
10
									 minutes
(B) Carbohydrate-containing media
12-15 minutes
(C) Contaminated test materials
30
									 minutes
(D) Membrane filtration units
15
									 minutes
(E) Sample containers
15 minutes
(F) Individual glassware
15 minutes
(G) Dilution water
15 minutes
(H) Rinse water
15-30 minutes
(vii) Records of each autoclave run shall be
						maintained in a laboratory notebook and include the date, contents,
						sterilization time and temperature, total cycle time (recorded as time in and
						time out) and initials of the responsible individual.
(viii) If an autoclave cycle fails to meet
						any requirement, corrective action shall be documented. Media may not be
						reautoclaved.