7 3 Vol. 7 No. 3 2016 3 Journal of Food Safety and Quality Mar., 2016 刘冬玲, 蒋 * 波, 陈奇知, 杨美成 (, 201203) 摘要 : 目的 方法 GB16740-2014, GB4789, 10 结果, 3 :, ;, 4, 结论,, 关键词 : ; ; ; Discussion on the method of microbiological test for health foods including Chinese herbal ingredients LIU Dong-Ling, JIANG Bo, CHEN Qi-Zhi, YANG Mei-Cheng * (Shanghai Institute for Food and Drug Control, Shanghai 201203, China) ABSTRACT: Objective To analyze the rationality of the methods of colony counting and specified micro-organisms in microbial tests for health foods including Chinese herbal ingredients. Methods According to GB16740-2014 Health food microbial limit requirements, Chinese Pharmacopoeia 2010 Appendix microbial limit tests and GB4789 Food microbiological analysis, microbiological test methods of 10 kinds of health foods including Chinese herbal ingredients (tea in solid form and oral liquid) were validated. Results During the methodology validation by conventional method, antibiotic effects of 3 samples on microbial enumeration tests could be eliminated by neutralizing method rather than dilute method. However, antibiotic effects of 4 samples on test of specified microorganism (Staphylococcus aureus) could not be eliminated by the test methods. Conclusion Method of neutralizing agar medium is recommended in microbial enumeration. The positive control strains are also added in the whole experimental process. More reasonable methods which are set up to detect microbe in health foods including Chinese herbal ingredients by reference to Chinese Pharmacopoeia should be validated. KEY WORDS: Chinese herbal ingredients; health foods; method validation; microbiological test * 通讯作者 :,,, E-mail: yangmeicheng@vip.sina.com *Corresponding author: YANG Mei-Cheng, Ph.D, Chief Pharmacist, Shanghai Institute for Food and Drug Control, Shanghai 201203, China. E-mail: yangmeicheng@vip.sina.com
888 7 1 引言, [1],,, [2,3],, [4-7] GB 4789 [8],, [9-12],,, [13], 10,, 2 材料与方法 2.1 材料和试剂 2.1.1 材料 10 ( 1): (: 4131001), (: 1130103), (: 131203), (: 130701); : ( : 13F05A01A), (: KB115), (: KSR581215), (: TLJ0570), (: 14D02B06B), (: TAK0604) 2.1.2 培养基 (: E0326Y)7.5% (: 3302004) ; (BS)(: 150508) ; (BPW, : 150420)(LST)( : 150320)(BGLB)(: 150522) ; (: 150526)0.85%(: 150526) (: 150526)Baird-Parker ( : 150527)(TTB)(: 150526)(XLD)(: 150526) 表 1 含中药成分的保健食品所含主要中药成分 Table 1 Chinese herbal ingredients in the tested health foods ()
3, : 889 2.1.3 菌种 (Escherichia coli) [CMCC(B) 44 102] (Staphylococcus aureus)[cmcc(b) 26003](Bacillus subtilis)[cmcc(b) 63501] (Candida albicans)[cmcc (F) 98 001](Aspergillusniger) [CMCC(F) 98003](Salmonella paratyphi B)[CMCC(B) 50 094], 2.2 实验方法 2010 GB 4789 2.2.1 菌液制备 : 1 ml 50~100 CFU : 1 ml 10~100 CFU 2.2.2 供试液制备 : 25 g 225 ml, 1:10 ; GB 16740-2014 [14], (), 25 g(ml) 225 ml 7.5% ( 225 ml );, 25 g(ml) 225 ml, 1:10,, 1:10, 2.2.3 抑菌性筛查试验 ( 常规法 ) : 1 ml (50~100 CFU) 1 ml, ;, 70%, : 10~100 CFU ;,,, 2.2.4 抑菌性消除试验 ( 培养基稀释法 中和法和两种方法联用 ), 70%,, : 1 ml 5 (0.2 ml/), 1 ml(50~100 CFU), 1 ml ;, 2.2.2 975 ml : 80 [13,15], ;, 3 结果和讨论 3.1 筛查抑菌性的结果 3.1.1 菌落计数 ( 常规法 ) 2, ( 2)10 7 70%, ; 2, 70% 0; 1, 1 70%,, 70% 0, 3 3.1.2 控制菌检查 ( 常规法 ) ( 3), 10 6, ; 4,, 4
890 7 Table 2 表 2 菌落计数的回收率 ( 常规法 %) Recovery of microbial enumeration tests (general method%) 1 2 1 2 1 2 1 2 1 2 80.0 86.1 0 7.0 0 4.4 109.5 85.1 77.6 83.6 71.8 90.3 0 1.17 0 2.94 98.4 89.6 75.9 73.8 91.9 82.2 89.7 86.1 86.4 82.5 91.6 86.0 82.8 98.3 90.7 81.2 85.2 94.4 75.3 86.0 79.5 75.4 86.2 93.3 100 82.2 102.4 108.3 103.9 82.5 97.7 75.4 93.1 88.3 101.5 92.1 56.0 77.8 1.30 0.00 95.3 100 77.6 93.3 103.1 94.1 90.5 75 81.8 91.2 97.7 94.7 87.9 98.3 92.3 80.2 71.4 75 98.7 96.5 95.3 80.7 81 76.7 98.5 75.2 91.7 84.7 83.1 78.9 81.4 89.5 93.1 88.3 96.9 79.2 97.6 101.4 80.5 91.2 94.2 91.2 93.1 101.7 Table 3 表 3 控制菌的验证结果 ( 常规法 ) Verification results of specified microorganisms (general method) (: ) 1:10 (10 ml) 1:10 (1 ml) 1:100 1:10(1 ml) 3.2 抑菌性消除的结果 3.2.1 菌落计数 ( 培养基稀释法和中和法 ) 3 ( 0.2 ml/)( 0.1% 0.7% 80 ) ( 4): 70%, 2 ;, 70%, 70%, 3.2.2 控制菌检查 ( 培养基稀释法 中和法和两种方法联用 ) 4 (7.5% 975 ml) 1( 0.1% 0.7% 80 225 ml7.5% ) 2( 0.3%3% 80 0.1% 225 ml7.5%)/ 2 ( 0.3%3% 80 0.1% 975 ml7.5%) ( 5): 4,
3, : 891 表 4 菌落计数的回收率 ( 培养基稀释法和中和法 %) Table 4 Recovery of microbial enumeration tests (dilute method and neutralizing method %) 1 2 1 2 1 2 1 2 95.1 92.9 95.1 77.9 97.4 87.3 73.4 74.6 90.2 97.6 95.1 87.0 96.2 76.2 95.3 76.1 88.4 93.8 0.0 0.0 100.0 85.1 85.1 95.5 Table 5 表 5 控制菌的验证结果 ( 培养基稀释法 中和法等 ) Results of test for specified microorganisms (dilute method and neutralizing method, etc) : 1: 0.1% 0.7% 80 2: 0.3% 3% 80 0.1% + 2 4 结论 10,, [4-7], (0.1% 0.7% 80),,,, 参考文献 [1]. [J]., 2011, 9: 111113. Jiang HT. Application study of edible and medicinal fungus in functional food [J]. Food Ind, 2011, 9: 111113. [2]. [J]., 2011, 32(21): 298300. Zhang B. Characteristics and safety of Chinese functional food materials [J]. Food Sci, 2011, 32(21): 298300. [3]. 1257 [J]., 2013, 23(2): 489491. Fu YS. Discussion on microbial contamination condition and impact factors of 1257 health food samples [J]. Chin J Health Lab Technol, 2013, 23(2): 489491. [4],,. [J]. (), 2005, 18(2): 5355. Cheng LL, Sun M, Tu L. Inhibition mechanism of extract of cassiaobtusifolial to plantpathogens [J]. J Sichuan Univ Sci Eng (Nat Sci Ed), 2005, 18(2): 5355. [5],,. [J]., 2014, 23(2): 9093. Li J, Zhang XS, Fu CS. The development of traditional Chinese medicine in antibacterial [J]. Chin Pharm, 2014, 23(2): 9093. [6] Cutler RR, Wilson P. Antibacterial activity of a new, stable, aqueous extract of allicin against methicillin resistant Staphylococcus aureus [J]. Br J Biomed Sci, 2004, 61(2): 14. [7],,. [J]., 2002, 13(4):196198. Qiu SC, Rong XG, Di DL, et al. The in vitro growth inhibition effect of coptis chinesis Frach (CCF) on bacteria [J]. Lishizhen
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