BIOLOGICAL INDICATORS      

   CERTIFICATE OF ANALYSIS   

Reorder No. SBA/6

Bacillus atrophaeus (Bacillus subtilis var. niger)              ATCC 9372

For:        Ethylene Oxide/Dry Heat Sterilization

Culture:  Soybean casein digest broth.

Storage: Controlled room temperature. Store away from sterilants. Do not desiccate.

Disposal: Incinerate or autoclave at 121°C for not less than 30 minutes.

Purity:   No evidence of contaminants using standard plate count techniques.

Lot No.: BSUB-270P

Manufacture Date: 2002 SEPTEMBER 09

Expiration: 24 months from Manufacture Date.

Heat Shocked Population:        2.1x10⁶        Spores/Unit

Assayed Resistance in glassine envelope:  

D-value reproducible only when exposed in an AAMI BIER vessel and cultured under the

exact conditions used to obtain results reported here. MPN and Survivor Curve methods

used.

Units are manufactured in compliance with SGM Biotech's quality standards, USP, EN 866,

ISO 11138 guidelines, and all appropriate subsections.

(1) D-value calculated using the Spearman-Karber method.

(2) Survival/Kill values are calculated according to USP and ISO 11138.

Biological Indicators to demonstrate adequacy of sterilization

INSTRUCTIONS

Sterilization

1. Place spore strips inside representative materials to be sterilized. Package or wrap as usual.

2. Locate test packages in areas of sterilizer most difficult to sterilize.  Identify test packages as to location In sterilizer.

3. After sterilizing, forward sufficient spore strips and products to sterility test laboratory along with at least one non-sterilized spore strip marked POSITIVE CONTROL.

Sterility Test Laboratory

1. All sterility testing should be performed in a clean, dust-free room - and within confines of a laminar flow hood. Personnel within the test area should be garbed in sterile gowns, masks, hoods, gloves, and boots. All transfers and manipulations must be conducted with rigid aseptic techniques to avoid "false positive" samples.

2. Procedure:

a.   Aseptically withdraw spore strips with sterile forceps and transfer to individual tubes
      containing 10-15 ml sterile soybean casein digest broth. Identify tubes.

b.  Incubate spore strips 7 days at appropriate temperature.  
    Ethylene Oxide/Dry Heat.........B. atrophaeus (B. subtllls var. niger) 30-35°C

c.   Observe tubes daily for growth.        Turbid = growth = non-sterile  
                                               Clear = no growth = sterile

Test Strip cultures should show no growth if sterilization has been achieved. If turbidity

and bacterial growth occur in test strip cultures, it suggests that the spores have

survived the sterilization process and are non-sterile.

3. Controls

a.   Positive: One or more positive controls should be included in each test series. Transfer a non-sterilized spore strip to culture medium and incubate with test series. Turbidity and growth indicates that the medium possesses suitable growth promoting qualities and that the spore strips contain viable spores. If positive control does not grow, do not use the lot.

b.   Negative: One or two tubes of culture medium incubated with test series. Absence of growth indicates that the medium was sterile prior to sterility testing.