BS EN 14561:2006 pdf download

BS EN 14561:2006 pdf download.Chemical disinfectants and antiseptics – Quantitative carrier test for the eva luation of bactericidal activity for instruments used in,the medical area- Test method and requirements (phase 2, step 2).
The interfering substance shall be sterile and prepared at 10 times its find concentration in the test.
The ionic composition (e.g. pH. calcium andlor magnesaim hardness) and chemical composition (e.g. mineral substances, protein, carbohydrates, lipids, detergents) shall be defined.
NOTE lithe following, the term ‘flederrg sttstanc& is sed even if it contaris more than one sitsiance.
5.2.2.8.2 Clean conditions (bovine albumin solution — low concentration)
— Dissolve 0,309 oF bovine albunwi fraction V (suitable for microbiological purposes) in 100 iN of cWuent
(5,2,2,4).
— Sterilize by membrane filtration (5.3.2.7), keep ina refrigerator (5.3.2.8) and iae within I month.
— The final concentration of the bovine albumin in the lest procedure (5.5) is 0,3 gIl.
5.2.2.8.3 DWty conditions (Mixture of bovIne albumin solutions — high concntratlon with sheep erythrocytes (se. 5,2.2.6))
Dissolve 3,00g of bovine atumin fracton V (suitable for microbiological purposes) in 97 ml of diluent (5.2.2.4). Sterilize by membrane filtration (5.3.2.7).
Prepare at least 8,0 nil fresh sterile defibrinated sheep blood (5.2.2.6). Centrifuge the sheep blood at 800 gN for 10 mm. After decarding the supernatant. resuspend ewyttvocytes in diuent (5.2.2.4). Repeat this procedure at least 3 times, until the supematant is colourtess, Resuspend 3 nil of the packed sheep erythrocytes in the 97 ml of sterited bovine albumin solution (see above). To avoid contamination this mixture should be split in portions probably needed per day and kept in separate containers for a maximum of 7 days in a refrigerator at 2 C to 8 C.
The final concentration of bovine albumin and sheep erythrocytes in the test procedure (5.5) shall be 3 gal and 3 mIll respectively.
5.3 Apparatus and glassware
5.3.1 General
Sterilize all gIassware and parts of the apparatus that will come into contact with the culture media and reagents or the sançle, except those which are supplied sterile, by one of the following methods:
a) by moist heat, In the autoclave [5.3.2.1 a)):
b) by dry heat ii the hot air oven (5.3.2.1 bfl
5.3.2 Usual microbiological laboratory equipmentZ
and in particular, the following:
5.3.2.1 Apparatus for sterilization:

b) for dry heat sterilization, a hot — oven capable of being malitained at (180 ,) ‘C for a minimum holding time of 30mm. at (170’‘5)”C for a mnmum holding time of 1 h or at (160’ J’C for a n’wnimtmi holdrng time of 2 h.
5.3.2.2 Water bathe, capable of bang controlled at 20D C ± I C, at 45 ‘c ± 1 ‘C (II pour plate technique is used – see 5.5.11) and at additional test temperatures ± I C (5.5.1).
5.3.2.3 Incubator, capable of being controlled at either 36 “C ± 1 ‘C or at 37 “C ± I ‘C. The same temperature shal be used for am rncubations performed during a lest and its controls and validation
5.3.2.4 pH-meter. having an inaccuracy of caAbration of no more thai ± 0.1 pH units at (20 ± 1 )‘C NOTE For meastsn9 the pH of the agar-media (52.2.3) a piaicture electrode or a Rat membrane electrode staId be used
5.3.2.5 Stopwatch
5.3.2.6 Shakers
a) Electromechanical agitator, e.g. Vode mixer31.
b) Mechanical shaker
5.3.2.7 Membrane filtration apparatus, constructed of a maleriat compatible with the substances to be filtered. with a titer holder of at least 50 ml volume, and suitable for use of filters of diameter 47 ram to 50 mm and 0,45 pm pore size for sterilization of hard water (52.2.7) and bovine albumin (5.2.2.8).
The vacuum source used shall give an even filtration flow ratt In order to obtain an uniform distribution of the microorganisms over the membrane and in order to prevent overlong filtration, the device shall be set so as to obtain the filtration of 100 ml of rinsing liquid in 20 s to 40s.
5.3.2.8 Refrigerator, capable of being controlled at 2 ‘C to 8 ‘C.
5.3.2.9 Graduated papettes of nominat capacities 10 it’d and I ml and 0.1 ml. Calibrated automatic pepettes may be used.
5.3.2.10 Path dishes (plates) of size 90 mm to 100 mm,
5.3.2.11 Glass beads (Diameter: 3 mm to 4mm).
5.3,2.12 Volumetric flasks.
5.3.2.13 Glass beads (diameter 0,25 mm to 0,5 mm)
5.3.2.14 Centrifuge (800 g,j)
5.3.2.15 Cylindrical plastic screw cap tttes. contents of about 15 ml, diameter about 18 mm (for the carrier).
5.3.2.16 Loop (metal or plastic).