Microbial Limit Test For Primary Packaging Materials – SOP

5.1 MICROBIAL LIMIT TEST OF PLAIN / PRINTED ALUMINIUM FOIL, GLASSIN PAPER, PVC FILM, POLY-  PROPYLENE BAGS BY CONTACT PLATE METHOD: :

• Clean the sampling polybags by wiping with 70 % v/v Iso Propyl Alcohol.
• Discard first 150 cm to 200 cm of foil / paper / film by cutting with the help of sterile scissor to avoid sampling of superficial contamination.
• Sample the required packaging material from sampling area as per Sampling, Testing and Approval/Rejection of Packaging Material – SOP. Label the packaging material with correct surface.
• Prepare SCDA and SCA as per Preparation and destruction of media for microbiological analysis-SOP and autoclave at 121°C  for 15 lbs for 15 minute.
• Transfer the plate under laminar flow bench.  Remove the lid of contact plate and pour sufficient agar media.
• Allow the media to get solidified then cover with the lid.
• Remove the sampled foil / paper / film from the polybags with sterile forceps.  Spread the foil / paper / film on LAF working bench in such a way that inner part which comes in contact with the product will be on the upper side.
• Remove the lid of the contact plate, invert the plate on foil / paper / film. Slightly press the plate from all the sides taking care to avoid the cracks in the  media.  Cover the plate with the lid and incubate at 30°C to 35°C for 5 days for bacterial count.
• Follow same procedure yeast and mould count as above using SCA media. Incubate SCA plates at 20°C to 25°C for 5 days for fungal count.

5.2 MICROBIAL LIMIT TEST OF  GLASS BOTTLE AND CAPS:

• Sterile Saline :  Prepare physiological saline ( 0.90 % NaCl in water ), sterilize at 121 °C for 15 lbs for 15 minute.
• Rinse the Glass Bottle and Caps with 5 ml of saline.
• Transfer the rinse saline in test tube approximate volume of 10 ml, mix it thoroughly and Transfer 1 ml of sample in sterile two Petri plate add 15 to 20 ml SCDA media for bacterial count and SCA media for fungal count respectively allow to solidify the media.
• Incubate the plates for 5 days, SCDA at 30°C to 35°C for bacterial counts and SCA 20°C to 25°C for  fungal counts and record result in cfu/ml for Glass bottle and caps.

5.3 TEST FOR PATHOGENS:

• Foil / Film / Paper: Cut approximately 10 x 10 cm with the sterile scissor, Inoculate this piece in SCDM and incubate at 30°C to 35°C for 24 hours for Enrichment then observe for the specified microorganisms as per Microbial limit test for finished products-SOP.
• Glass Bottle and Caps: Transfer the left over (as in MLT limit test of glass bottle and caps) saline in 90 ml of SCDM and incubate at 30°C to 35°C for 24 hours for Enrichment then observe for the specified microorganisms as per Microbial limit test for finished products-SOP.

5.4 ACCEPTANCE CRITERIA FOR APPROVAL :

• For Plain / Printed aluminium foil, Glassin Paper, PVC film, Poly- propylene bags :
  • Total Aerobic Bacterial Count : NMT 250 cfu/10cm x 10cm of material.
  • Total Yeast and Mould Count  : NMT 100 cfu/10cm x 10cm of material.
  • Pathogens : Absent.
• For Glass Bottle and Caps :
  • Total Aerobic Bacterial Count : NMT 250 cfu/ml.
  • Total Yeast and Mould Count  : NMT 100 cfu/ml.
  • Pathogens : Absent.

5.5 FREQUENCY :

• Initial three supply from new manufacturer.
• Quarterly, irrespective of Product / Specification.