为探究ATP生物荧光增幅法在化妆品微生物检测中应用的可行性,开展了富集培养基促生长能力测试和产品适用性试验。收集了与化妆品原料及其生产环境相关的47株代表性菌株,在富集培养基MTAT中生长良好,ATP生物荧光增幅检测结果均为阳性。分别选取了清洁类、护理类、美容修饰类中6种化妆品,接种6株《中国药典》(2020版)参考菌株,进行了产品适用性试验。样品影响测试结果表明:培养基空白的信号值在167~285 RLU, 6种产品的本底信号值在87~199 RLU,远低于3倍培养基空白信号值的要求;ATP回收率在73.8%~106.8%,均高于25%的推荐值,符合检测方法的要求。接菌试验结果表明,供试样品的检测结果均为阳性,化妆品的抑菌作用可被有效消除。因此,ATP生物荧光增幅法具有在化妆品及相关领域开展微生物快速筛选与检测的良好应用前景。
翟磊
,
葛媛媛
,
洪海军
,
刘吉泉
,
李婷
,
沈颖
,
林雅芳
,
蔡俊松
,
刘骥
,
刘瑞娜
,
王旭
,
姚粟
. ATP生物荧光增幅法在微生物检测中的可行性研究─应用于化妆品领域[J]. 食品与发酵工业, 2020
, 46(21)
: 201
-206
.
DOI: 10.13995/j.cnki.11-1802/ts.025092
To explore the feasibility of amplified ATP bioluminescence method for microbial detection in cosmetics, growth promotion test of an enrichment medium and product suitability test were carried out. Forty-seven representative strains relevant to raw materials and production environment of cosmetics were collected and tested. They all grew well in the enrichment medium, and the detection results of amplified ATP bioluminescence were all positive. Subsequently, six kinds of cosmetics from cleanser, care cosmetics and makeup categories were selected and inoculated with six reference strains recommended by Chinese Pharmacopoeia (2020) for product suitability test. Results of sample effect test showed that the signal value of medium blank was between 167-285 RLU, while the background signal value of 6 products was between 87-199 RLU, which was far below the requirement of 3-fold of medium blank signal value. ATP recovery ranged from 73.8% to 106.8%, both higher than the recommended value of 25%, which met the requirements of the detection method. Results of spiking test showed that all the samples tested were positive, proving efficient elimination of antimicrobial effect of cosmetics. Therefore, amplified ATP bioluminescence assay has a promising application prospect in the rapid screening and detection of microorganisms in cosmetics and related fields.
[1] 李青彬,赵晓军,王香,等.化妆品微生物污染状况及其分析[J].中国卫生检验杂志,2006,16(2):245-247.
[2] 谢小保,欧阳友生,陈仪本,等.化妆品微生物污染状况研究[J].中国卫生检验杂志,2005,15(9):1 105-1 105.
[3] 郑萍,陈西平.化妆品微生物污染情况调查[J].中国卫生检验杂志,2009,19(10):2 405-2 406.
[4] 郑萍,陈西平.化妆品的微生物污染问题及防腐效能评价方法[J].中国卫生检验杂志,2007,17(11):2 122-2 124.
[5] 化妆品安全技术规范(2015年版)[S].北京:国家食品药品监督管理总局,2015.
[6] 文霞,杨秀茳,谢小保.化妆品微生物检测技术的研究进展[J].日用化学工业,2015,45(2):110-114.
[7] 陈倩,陈昭斌.微生物快速检验方法新进展[J].中国卫生检验杂志,2016,26(7):1 061-1 064.
[8] 郝良玉,曲晗,李志萍,等.微流控技术在病原微生物检测中的应用[J].检验医学与临床,2018,15(21):3 299-3 302.
[9] 杜美红,赵瑞雪,李静雯.基质辅助激光解吸电离飞行时间质谱分析技术在微生物检测与鉴定中的应用[J].食品安全质量检测学报,2017,8(2):457-461.
[10] 董金颖,张阳,刘皓,等.拉曼光谱技术在病原微生物快速检测中的应用与展望[J].中华检验医学杂志,2018(1):5-8.
[11] ISO 16140-2,Microbiology of the Food Chain-Method Validation Part 2:Protocol for the Validation of Alternative (Proprietary) Methods Against a Reference Method[S].International Organization for Standardization,2016.
[12] AOAC INTERNATIONAL Methods Committee Guidelines for Validation of Qualitative and Quantitative Food[S].Association of Official Analytical Chemists,2002.
[13] 《中国药典》[M].2020版.北京:国家食品药品监督管理总局,2020.
[14] 郑小玲,王银环,陈君豪,等.药品微生物检验替代方法国内外研究进展[J].药物分析杂志,2020,40(4):577-582.
[15] 白飞荣,葛媛媛,蔡俊松,等.化妆品中霉菌和酵母菌检验MSDA方法的验证研究[J].日用化学品科学,2019,42(6):40-47.
[16] SCOTT SPAETH,QUYNH TRAN,ZHIYUN LIU.Evaluation of an ATP-bioluminescence rapid microbial screening method for in-process biologics[J].PDA Journal of Pharmaceutical Science and Technology,2018,72(6):574-583.
[17] 魏树源,杨京生,陈晓琦,等.三磷酸腺苷生物发光快速微生物检测法在疫苗中间无菌试验中的应用[J].中国生物制品学杂志,2010,23(10):1 120-1 124.
[18] LIEBERS V,BACHMANN D,FRANKE G,et al.Determination of ATP-activity as a useful tool for monitoring microbial load in aqueous humidifier samples[J].International Journal of Hygiene and Environmental Health,2015,218:246-253.
[19] 路福平,闫青松.化妆品产品微生物快速检测-ATP快速荧光法[J].中国洗涤用品工业,2004(5):35-36
[20] GB/T 18670—2017化妆品分类[S].北京:中国标准出版社,2017.
