살균소독제 안전성유효성 평가 Components of Antimicrobial Efficacy 한국보건산업진흥원 조 양 희 살균소독제 안전성유효성 평가 Components of Antimicrobial Efficacy Formula Test Method Use Condition 한국보건산업진흥원 조 양 희

연구 배경 식중독 대형화 추세 다양한 살균소독제 개발 식품에 간접적으로 잔류 살균, 소독, 멸균 등 효능 표시 : 소비자 인식 미생물로 인한 부패 또는 변패 방지 관심증가 다양한 살균소독제 개발 개발 요구 증가 및 선진 외국 제품 개발 무역 마찰 요인 식품에 간접적으로 잔류 안전성 요구 살균, 소독, 멸균 등 효능 표시 : 소비자 인식 유효성 평가 요구

관련 문헌 고찰 국내 시장 현황 연구추진 체계 안전성 평가지침 유효성 평가지침 안전성 평가기준 표시 기준 유효성 평가기준 제품 유형 표시 사항 독성 자문위원 제품 적용 범위 안전성 평가지침 유효성 평가지침

식중독 주요 원인 균 비교 순위 일본 한국 미국 1 Salmonella 2 Vibrio Staphylococcus Clostridium 3 E. Coli 4 5 Bacillus

관련근거 : 식품위생법 제7조제2항, 제9조제2항 및 시행규칙 제4조제1항 살균소독제 인정제도란? 제2005-75호 배경 : 식품위생법 개정 (2002. 8. 26 공포, 2003. 8. 27 시행) 신설 관련근거 : 식품위생법 제7조제2항, 제9조제2항 및 시행규칙 제4조제1항 살균소독제 인정제도란? 제2조(정의)제2호 “식품첨가물”이라함은 식품을 제조·가공 또는 보존함에 있어 식품에 첨가·혼합·침윤 기타의 방법으로 사용되는 물질(기구 및 용기·포장의 살균·소독의 목적에 사용되어 간접적으로 식품에 이행될 수 있는 물질을 포함한다.)을 말한다. 기구 등의 살균소독제 제품은 유효성분 및 보조성분의 종류 및 배합비율에 따라 여러종류의 제품 조합이 가능하므로 개개의 제품별로 사용기준에 따른 유효성과 안전성을 검토하여 인정하는 제도임(“기구등 살균소독제의 한시적 기준 및 규격인정”) 살균소독제 제품이란? 일반주방업소, 단체급식업소 등에서 사용하는 식기류, 식품의 가공기구 조리기구, 식품공장의 제조 기기 및 설비 등에 사용되어 유해미생물에 대해 살균소독 작용을 하는 제품으로 사용목적에 따라 살균소독제를 처리한 후 헹구지 않고 사용되거나 음용에 적합한 물로 헹구는 제품 살균소독제 인정제품(2007.3.31) : 총 152제품

관련규정 평가방법 AOAC EU-BPD Biocides 유럽연합 CEN EPA-FIFRA APs 미 국 한 국 m-CEN* 미 국 한 국 식품위생법 살균소독제 m-CEN*

Non-public health products Control growth of algae, odor-causing bacteria, bacteria which cause spoilage, deterioration or fouling of materials and microorganisms infectious only to animals Cooling towers, jet fuel, paints, and treatments for textile and paper products Public health products Control microorganisms infectious to humans in any inanimate environment Used to destroy or eliminate all forms of microbial life including fungi, viruses, and all forms of bacteria and their spores. Spores are considered to be the most difficult form of microorganism to destroy. 1. Sterilizers (Sporicides) Hospitals (medical and surgical, instruments and equipment)

Public health products Used on hard inanimate surfaces and objects to destroy or irreversibly inactivate infectious fungi and bacteria but not necessarily their spores. 2. Disinfectants Hospital use General used (households, swimming pools, and water purifiers) 3. Sanitizers Used to reduce, but not necessarily eliminate, microorganisms from the inanimate environment to levels considered safe as determined by public health codes or regulations. Carpet, Air, Laundry additives, in-tank toilet bowl Non Food Contact Sanitizers Food Contact Sanitizers

Public health products 3. Sanitizers Used to reduce, but not necessarily eliminate, microorganisms from the inanimate environment to levels considered safe as determined by public health codes or regulations. Dish, cooking utensils, equipment and utensils found in dairies, food-processing plants etc. Non Food Contact Sanitizers Food Contact Sanitizers 4. Antiseptics and Germicides Used to prevent infection and decay by inhibiting the growth of microorganisms. On living humans or animals Drug FDA

Bacillus subtilis (ATCC 19659) and Clostridium Sporogenes (ATCC 3584) Level of Activity Test Method, AOAC Test Microorganisms Total No.of Carriers Performance Standard STERILIZE-RS Sporicide Test Method Bacillus subtilis (ATCC 19659) and Clostridium Sporogenes (ATCC 3584) Killing on all 720 carriers 720 60 carriers X 2 type of surface X 3 samples X 2 type of test organism DISINFECTANTS Use-Dilution (solids & liquids) or Germicidal Spray (sprays) Salmonella choleraesuis (ATCC 10708) or Stapylococcus aureus (ATCC 6538) Limited Efficacy Must kill 59 out of each 60 carriers 180 60 carriers X 1 type of surface X 3 samples X 1 type of test organism

Use-Dilution (solids & liquids) or Germicidal Spray (sprays) Level of Activity Test Method, AOAC Test Microorganisms Total No.of Carriers Performance Standard Use-Dilution (solids & liquids) or Germicidal Spray (sprays) General or Broad Spectrum Efficacy Salmonella choleraesuis (ATCC 10708) and Stapylococcus aureus (ATCC 6538) Must kill 59 out of each 60 carriers 360 60 carriers X 1 type of surface X 3 samples X 2 type of test organism Hospital or Medical Environment Efficacy Use-Dilution (solids & liquids) or Germicidal Spray (sprays) Salmonella choleraesuis (ATCC 10708) and Stapylococcus aureus (ATCC 6538) and Pseudomonas aeroginosa (ATCC 15442) Must kill 59 out of each 60 carriers 540

Enterobacter aerogenes (ATCC 13048) Level of Activity Test Method, AOAC Test Microorganisms Test Condition Performance Standard SANITIZERS For Nonfood Contact Surfaces USEPA’s Test Method (91-30,D8) Stapylococcus aureus (ATCC 6538) and Klebsiella pneumoniae (ATCC 4352) (e) One test on each represen-tative surface 99.9% reduction compared to control within 5 minutes Enterobacter aerogenes (ATCC 13048) For Food Contact Surfaces ASTM E1153-03 * For Halides Available Chlorine Germicidal Equivalent Concentrat-ion Method Salmonella typhi (ATCC 6539) or Stapylococcus aureus (ATCC 6538) One test on each of 3 samples Concentration equivalent to 50, 100 and 200 ppm available chlorine 3 different batch ( 1 : 60days old)

Level of Activity Test Method, AOAC Test Microorganisms Total No.of Carriers Performance Standard For Food Contact Surfaces * For Other Products Germicidal and Detergent Sanitizers Method Escherichia coli (ATCC 11229) Stapylococcus aureus (ATCC 6538) One test on each of 3 samples 99.999% reduction in each test organism within 30 seconds Reference : USEPA Pesticide Assessment Guidelines Subdivision G : Product Performance, Part 91-2.

Standard Operating Procedure (SOPs) A part of the Antimicrobials and Plant Pathogens Branch of the Biological and Economic Analysis Division (BEAD) Standard Operating Procedure (SOPs) SOPs for antimicrobial testing methods to determine the efficacy of hard surface disinfectants Antimicrobial Testing Methods & Procedures (ATMP) Methods 11 ID Numbers (10 open) Procedures 37 ID Numbers (34 open) * OPP : Office of Pesticide Programs

Ch- 6. Disinfectants Subchapter 1 : PHENOL COEFFICIENT METHODS(3) Emma Singleton, Chapter Editor, U.S. Food and Drug Administration Subchapter 1 : PHENOL COEFFICIENT METHODS(3) Test Disinfectants against Sallmonella typhi (ATCC 6539) Phenol Coefficient Method First Action 1955 Final Action 1964 AOAC Official Method 955.11 EPA PR(Pesticide Registration) Notice 2001-4 (2001/6/19) FIFRA SAP (Scientific Advisroy Panel) “Elimination of Phenol Resistance Testing for Antimicrobial Disinfectant and Sanitizer Pesticides” First Action 1955 Final Action Test Disinfectants against Staphylococcus aureus (ATCC 6538) Phenol Coefficient Method AOAC Official Method 955.12 First Action 1955 AOAC Official Method 955.13 Test Disinfectants against Pseudomonas aeruginosa (ATCC 15442) Phenol Coefficient Method

Subchapter 2 : HARD SURFACE CARRIER TEST METHODS(6) AOAC Official Method 955.14 Testing Disinfectants against Salmonella choleraesuis (ATCC 10708) Use-Dilution Method First Action 1955 Final Action 1959 Repealde F.A 1988 Surplus 1993 AOAC Official Method 955.15 Testing Disinfectants against Staphylococcus aureus (ATCC 6538) Use-Dilution Method First Action 1955 Final Action 1959 Repealde F.A 1988 Surplus 1993 AOAC Official Method 964.02 Testiing Disinfectants against Pseudomonas aeruginosa (ATCC 15442) Use-Dilution Method First Action 1964 Surplus 1993 AOAC Official Method 991.47 Testing Disinfectants against Salmonella choleraesuis (ATCC 10708) Hard Surface Carrier Test Method First Action 1991 AOAC Official Method 991.48 Testing Disinfectants against Staphylococcus aureus (ATCC 6538) Hard Surface Carrier Test Method First Action 1991 AOAC Official Method 991.49 Testing Disinfectants against Pseudomonas aeruginosa (ATCC 15442) Hard Surface Carrier Test Method First Action 1991

“Andistand” (SMT4-CT98-2222) Project – Produce Standards (Test Methods) – Antimicrobial efficacy of Chemical Disinfectant and Antiseptics (published : 15, Development : 18) The Development of Test Methods Horizontal Working Group - Harmonization Task Group - Ring Trial Task Group – 4 Working Groups(WGs) HWG Basic test method (Phase 1), Ring trial tests 3(4) WG1 Human Medicine market 5(20) WG2 Veterinary market 3(8) WG3 Food Hygiene, Domestic and Institutional market 5(7) “Andistand” (SMT4-CT98-2222) Project : Inter-, and Intra-laboratory Ring trial tests

“Disinfects Surface” or “Surface disinfectant” Phase Test Type Standard Test Method Phase 1 Quantitative suspension test under “clean” conditions for basic activity 105 (B) EN 1040 104 (F) EN 1275 104 (S) prEN 14347 Phase 2 Step 1 Quantitative suspension test under conditions representative of practical use EN 1276 EN 1650 103 (S) EN 13704 103 (V) EN 13610 Step 2 Quantitative surface test simulating practical conditions 104 (B) EN 13697 103 (F) Phase 3 Field test under practical condition “No Claim” “Disinfectant” “Disinfects Surface” or “Surface disinfectant”

US-EPA (AOAC) EU-CEN (EN) 960.09(GDS) 955.16(ACE) Phase 1 P 2-Step 1 기 준 99.999% Same 99.999% Staphylococcus aureus (ATCC 6538) and Escherichia coli (ATCC11229) Salmonella typhi (ATCC 6539) or Staphylococcus aureus (ATCC 6538) Escherichia coli(ATCC 10536) Staphylococcus aureus(ATCC 6538) Pseudomonas aeruginosa(ATCC 15442) Enterococcus hirea(ATCC 10541) 시험균 시험조건 25°C/30s 20°C/1min 20°C(4,10,40)/5min(15,30,60) 간섭물질 Organic soil(optional) Bovine Albumin 외 5종 중화제 중화배양액(optional) 기본중화혼합제 외 15종 정량 Suspension 비교 Suspension 정량 Suspension 정량 Suspension 특징 GDS: Germicidal & Detergent Sanitizer ACE: Avaliable Chlorine Germicidal Equivalent Concentration 비고 P2-Step1 : Phase 2 Step1

ITEM CEN EN 13697 (2001) ASTM QCT-1 (2000) ASTM QCT-2 (2002) CARRIERS Stainless Disk Glass Vial SOIL LOAD BSA Alone Two Level Tripartite Mix One Level REPLIACTES ??? 5-10 Test & 3 control INOCULUM 50㎕ 10 ㎕ TEST AGENT 100 ㎕ /Carrier 50 ㎕ /Carrier ELUTION Rotation on Beads Votex/Sonication CFU ASSAY -20% Eluate Pour plate Entire Eluate, Membrane Filter PERFORMANCE CRITERION 4 log10 4log10 - 6log10

QCT 시험방법 비교 시험법 항목 ASTM E2111 E2197 CEN 13697 AOAC UDM HST Method Type QHST QCT Carrier Type GV SD SC GC Test Microbes B(3)/F(1)/S(2) B(3)/F(1)/S(2)/V(4) B(3)/F(2) B(3) Soil load TSL BSA/2S - IV/SV 10/50㎕ 50/100㎕ 0.5/10㎖ Test Condition optional RT/5m 20℃/10m R/C MF PC G Standard 4-6 log 4 log NG/360

살균소독제 유효성평가 방법 다양화 Harmful Microbes Disease Surrogate Microbes Coronavirus SARS Poliovirus type 1 (Sabin vaccine strain) HIV AIDS norovirus Food poisoning Bacillus anthracis Antrax Bacillus atrophaeus Mycobacterium tuberculosis Tuberculosis Mycobacterium terrae

HSCT(Hard Surface Carrier Test) Method 1996년 Pesticides Programme 하에 설치 목표 1. 회원국간 자료 요구의 조화 2. 유효성 시험과 허용한계치 선정 3. 환경상 거동과 영향에 대한 시험지침 개발 (인체건강 및 생태계영향) 4. 노출평가와 위해성 평가지침 개발 5. 각 회원국간의 biocide 심사 및 평가의 조화 6. Biocides로 인해 발생가능한 위해성 저감 TFB(Task Force on Biocides) : 2003. 7 1. Efficacy Work 2. Environmental Exposure Assessment