‘MicnopioLocteaL MeTHO0S Chapter 17,9. 114 subchapter 6 ss STERILITY (COMMERCIAL) oh Fao (CANNED, LOW ACI rsot ‘AOAC Official Method 972.44 ‘Microbiotogical Method First Action 1972 Final Action 1978 mustaches, or sideburns bow car 1006 unless these are completely ar clean laboratory coat For (Personnel with beards, stoulznapetem seityeamnsic ovr ih stereos snd as. Wes cmintn) A Pine “Low acid foods” means any food with finished equilibrium: pute te Met pls ny 1 comuines wc show 0 San ier cl eat cnines 10 dys at 2-35°C remit ety slo a sno aoe by option fat wih edo fod feof vil forms of ror avn phe hath sigan, a wel 5 cerns nt of het sifcance eb of wodcing inte te nd noma eeiptedonis osrge and Statin Muto and Rages 100, spi L110, eto panda een in Erlenmeyer. Asepially pour ito str es 'y pour into sterile Peti dishes and allow and 15.0 agar saan, 9 B58 INL HO, beat (©) Detergent sanitizer soltion.—pHi oe PHisoDerm, orequivalent (9 Con pera © 2016 AOAC INTE} _ ‘TIONAL (AcOEt —_pispossble, operating FOOM-19 (Baxi (©) Cops Die Gaw Pat IL, USA, or equi SO ee aight wall, 200-250 mm long » 7 my p. Test Sampling id avi ttt in cean room (HF PECESEY, On oon wed pat beeline) fave, we La Srp abel from cas, examine cans for ext gy” can descriptions, Washeans wih so2 (0 erp sa so yan 1,0, and ry wih CEI PE O¥Es, Wipe say 100 pm (up Cl sltion (eB, Coe te solution immediatly befor placing Washed and dng ea cece code end of can in down position and number ca. ra wth CuSO, marking solution to ight of kde sean sr ands and face with Soap, and revash bands ao win iergent sanitizer oution. Completely cover aii ca able operating room cep. blue portion of lame, Heat this end of can untill cn ed then return canto table informer is evapora f tandte and blade of special can opener, Cla), with pepe tw moistened with 70% alcohol, flame metal portion enou destroy all microorganisms, and use it to make 4 om (15 diameter hole in noncoded, heated end of can. Immedia ‘without moving can, use straight-wall sterile glass pipet, Cle) transfer ca2 g food to separate tubes, two each of aerobic and tw of anaerobic media (four total). (No other transfer be substituted) Preloosen screw cap and hold it between i ring fingers while transfer is being made. Fame lips of media both before and after addition of food. When transferring food anaerobic tubes, food must be inoculated into lower port medium. Tighten screw caps after inoculation, incubate tutes 2 at 35°C, and observe daly. Record results for each tube separ! Remove additional 210 g food test portion from exch cnt with sterile pipet and place in sterile 25 * 200 mm screw lest tube. Use pipet-like spatula, if necessary, for this ope (thermophilic contamination unlikely). Number tube to corspo! ‘o can and refrigerate for late testing, if necessary Contamination Control Use sterile loop org : ‘0r glass rod to streak plate of glucose starch 'B(e). On table, open plate of glucose starch agar for time ea Hongest duration hat any medium tube or plate is exposed. nab? Plates 72-hat 35°C, and observe daly F Microscopic Examination au ‘Pair of metal cuting shears, enlarge hole in can ade" ome Meemscopelly (oil immersion) examine heat fuel Shed stained 10s with 1% gentian (or crystal) vk washed in 2 wand it tnning tp water, or, atematively, examine ¥e°— Pol Rue Coast microscope. If food contain apres rr gould be dripped ‘across food smear while itis still pot fis heat fixing. ¢ ce ‘ompare stained smear with one made from ™ ©: pH Determination Deter 3! PH of foe with pH meter, using reference bufer ea Compare 1g °°22%d both reference buffer pH and tests normal can of food, if available.1S OF ANALYSIS (2016) sornaion of ROSS C00 ry storms ot a0eMSlAPPEANNE,sboomal pH aise Ton mizosopic examination, andor growth in oan of fod, subculture correspondin ane ip OF tbe and, with aie ET ganser 2 food 1 0 tubes each of aerobic Fe ooo aly Rane He of mes tog sadn eration of food. Tighten caps after inoculation, eas 2 Ha 88°C and observe ily, Record resus for see seael cote isolated from normal cans heving obvious eae thc pode 0 in anaerobic medium 38°C should woth spect as sing fo boraory contamination, fey eda ing nnn ino another normal can, ewe an incubate 14 days at 39°C. Any sweling 13 mm) (machined in local machine shop) with No.2 taper in piercer top, beveled 1/8 3/16 in. (3*5 mm) at bottom V4 % 1/2 in. (6 * 13 man) silicone rubber gasket around beveled 1/8 in. 3 mm) piercer to maintain sea. (© Helium exposure tank—ASME paint tank, 19 gal G7B 1), tested to 100 psi (690 kPa), equipped with inlet and ‘outlet microcontol valves (Harrison Industral Technologies, Ine, Fifield, OH, USA). (@) Pressurized helium tank —With 2-stage regulator. (©) Timer and solenoid —To automate release of He from exposure tank. (Helium gas standards Scott Specialty Gases (South Pinfield, NI, USA). (&) Cyanoaerylate glue —SuperGlue (Adhesive System, 3M, Industrial Tape and Specialties Division, 3M Center, St Paul, MN, USA, or equivalent) (bh) Can opener-—Bacteriological, 972.44C\a) (see 17.6.0). (@) Rubber dises 2 38» 18 in. (60 «3 mm) and 70 durometer (Netherland Rubber Co, Cincinnati, OH, USA), ©. Calibration Test Procedure For gas chromatographs equipped with side port loop (0.5 mL), inject 5.0 mL calibrated He standards (suggested range of 5, 15, 25, 50, and 75% He). For instruments not equipped with side port loop, inject appropriate volume of standards. Use same volume for analysis of headspace gas test portions. Plot percent He versus He peak height at attenuation used. Depending on quality of instrument, plot should approximate a straight line. ‘Check gauge on can piercer against known pressure and vacuum. ‘Test resealing procedure, E, on control cans. . Helium Exposure Tank Control introduction rate of He into exposure tank and time ‘cans are exposed to He pressure at 45 £ 2 psi (310.5 + 13.8 kPa). “Timer, solenoid, and microvalves with vernier scales can facilitate procedure, Connect He source to exposure tank. Tum timer on to close outlet solenoid valve. Approximately 15-20 min are needed to reach 45 psi (310.5 kPa) in tank. Make minor adjustments if necessary. Adjust timer to expose cans to He pressure at 45 psi {G10.5 kPa) for 30 min (30 min exposure period is in addition to time necessary to reach 45 psi (310.5 kPa)]. Tank pressure should bbe reduced to 0 psi (KP) within 5-10 min, . Preparation of Can for Helium Test It test portion is tobe taken for microbiological testing, proceed asin 972.440 (see 17.601). (© 2016 AOAC INTERNATIONAL