基于双培养体系的化妆品微生物ATP生物荧光增幅法适用性研究

doi:10.3969/j.issn.2097-2806.2025.07.012

Abstract

Abstract:

In order to evaluate the suitability of the amplified ATP bioluminescence assay based on two cultivation systems in the rapid screening of cosmetic microorganisms, 51 commercially available cosmetic products covering 20 domestic and foreign brands, including different product types, functional claims, action sites, product dosage forms, and user groups, were widely selected for method suitability research with reference to the method suitability test requirements of sterility inspection method and microbial counting method in the Chinese Pharmacopoeia （2020 Edition）. And the suitability of the method was evaluated through sample impact test and spiking test results. The results show that 45 of the 51 products have a background fluorescence signal value less than 3-fold the signal value of blank medium, which meet the requirements of the sample impact test. While six products failed the sample impact test. Six representative strains are inoculated in 45 products that meet the requirements of the sample impact test according to the final concentration of 10-100 CFU/g, and are positively detected in 42 products after being cultured in the two cultivation systems. While some strains in three test products do not get positive detection results. In conclusion, the amplified ATP bioluminescence assay based on two cultivation systems has good suitability in the rapid screening of microbial contamination samples in cosmetics, and provides an efficient and reliable new method for microbial safety control in the whole process of cosmetics production.

Key words: two cultivation systems, amplified ATP bioluminescence assay, cosmetics, microorganism, suitability

CLC Number:

TQ658

Ruina Liu, Biao Jiang, Yafang Lin, Lizhe Yang, Lei Zhai, Kun Li, Su Yao. Suitability study of amplified ATP bioluminescence assay for microbial detection of cosmetics based on two cultivation systems[J].China Surfactant Detergent & Cosmetics, 2025, 55(7): 909-919.

Figures/Tables 3

Tab. 1

Tab. 2

Sample effect test results of the tested cosmetic products"

编号	产品名称	批次	培养基空白（Avg. RLU）	微生物对照RLU/培养基空白RLU	ATP培养基对照（Avg. RLU）	产品样品（Avg. RLU）	平板生长情况	微生物样品RLU/培养基空白RLU	ATP样品（Avg. RLU）
1	清润黑面膜	Lot 1	124	P	6 344	143	-	P	5 828
		Lot 2				158	-	P	5 974
		Lot 3				163	-	P	5 743
2	氨基酸泥浆精华面膜	Lot 1	124	P	6 344	1 006 209	+	P	99 999 999
		Lot 2				2 028 105	+	P	99 999 999
		Lot 3				2 193 980	+	P	99 999 999
3	角质清洁面膜	Lot 1	124	P	6 344	210	-	P	6 561
		Lot 2				141	-	P	6 459
		Lot 3				295	-	P	5 957
4	清洁面膜	Lot 1	124	P	6 344	159	-	P	5 571
		Lot 2				191	-	P	5 596
		Lot 3				177	-	P	5 557
5	卸妆膏	Lot 1	124	P	6 344	326	-	P	5 750
		Lot 2				246	-	P	5 839
		Lot 3				187	-	P	5 801
6	精华定妆喷雾	Lot 1	124	P	6 344	156	-	P	5 607
		Lot 2				137	-	P	5 933
		Lot 3				135	-	P	5 708
7	修容粉饼	Lot 1	124	P	6 344	1 998 642	+	P	167 326
		Lot 2				2 030 490	+	P	173 745
		Lot 3				3 204 354	+	P	130 474
8	维生素乳液	Lot 1	180	P	4 765	215	-	P	3 175
		Lot 2				174	-	P	3 040
		Lot 3				204	-	P	3 237
9	保湿滋润乳	Lot 1	127	P	5 205	125	-	P	3 819
		Lot 2				118	-	P	4 100
		Lot 3				197	-	P	3 893
10	美容洗面奶	Lot 1	180	P	4 765	213	-	P	4 830
		Lot 2				148	-	P	4 093
		Lot 3				170	-	P	4 555
11	保湿洗面奶	Lot 1	180	P	4 765	116	-	P	3 263
		Lot 2				140	-	P	2 642
		Lot 3				125	-	P	2 681
12	保湿舒缓修护乳	Lot 1	180	P	4 765	239	-	P	5 604
		Lot 2				171	-	P	5 305
		Lot 3				164	-	P	5 025
13	倍护润体乳	Lot 1	180	P	4 765	11 542	-	P	16 565
		Lot 2				8 048	-	P	12 813
		Lot 3				8 198	-	P	13 260
14	舒缓柔嫩保湿护手霜	Lot 1	180	P	5 205	24 556	-	P	31 798
		Lot 2				34 104	-	P	44 846
		Lot 3				33 079	-	P	46 275
15	水凝保湿霜	Lot 1	180	P	4 765	1 923	-	P	6 423
		Lot 2				3 153	-	P	8 100
		Lot 3				3 170	-	P	8 512
16	水感多效防晒露	Lot 1	180	P	4 765	174	-	P	3 991
		Lot 2				178	-	P	3 960
		Lot 3				154	-	P	4 902
17	抗皱修护晚霜	Lot 1	127	P	5 205	159	-	P	5 519
		Lot 2				162	-	P	5 171
		Lot 3				201	-	P	5 119
18	泡泡洗手液	Lot 1	120	P	4 624	107	-	P	4 048
		Lot 2				103	-	P	3 946
		Lot 3				126	-	P	4 464
19	宝宝爽身粉	Lot 1	120	P	4 624	163	-	P	5 105
		Lot 2				107	-	P	5 135
		Lot 3				148	-	P	4 784
20	修容蜜粉	Lot 1	120	P	4 624	3 173 657	+	P	99 999 999
		Lot 2				3 188 422	+	P	99 999 999
		Lot 3				3 158 892	+	P	99 999 999

Tab. 2

Tab. 3

Spiking test results of the tested cosmetic products"

编号	产品名称	批次	培养基空白（Avg. RLU）	微生物对照RLU/培养基空白RLU						产品样品（Avg. RLU）	平板生长情况	微生物样品RLU/培养基空白RLU
编号	产品名称	批次	培养基空白（Avg. RLU）	黑曲霉	白色念珠菌	枯草芽胞杆菌	铜绿假单胞菌	金黄色葡萄球菌	大肠埃希氏菌	产品样品（Avg. RLU）	平板生长情况	黑曲霉	白色念珠菌	枯草芽胞杆菌	铜绿假单胞菌	金黄色葡萄球菌	大肠埃希氏菌
1	精华修护身体乳	Lot 1	229	P	P	P	P	P	P	250	-	P	P	P	P	P	P
		Lot 2								196	-	P	P	P	P	P	P
		Lot 3								179	-	P	P	P	P	P	P
2	爽肤香氛沐浴乳	Lot 1	229	P	P	P	P	P	P	234	-	P	P	P	P	P	P
		Lot 2								212	-	P	P	P	P	P	P
		Lot 3								287	-	P	P	P	P	P	P
3	滋养美肤沐浴乳	Lot 1	162	P	P	P	P	P	P	113	-	P	P	P	P	P	P
		Lot 2								106	-	P	P	P	P	P	P
		Lot 3								106	-	P	P	P	P	P	P
4	密集修护发膜	Lot 1	178	P	P	P	P	P	P	277	-	P	P	P	P	P	P
		Lot 2								240	-	P	P	P	P	P	P
		Lot 3								207	-	P	P	P	P	P	P
5	头皮护理润发精华素	Lot 1	178	P	P	P	P	P	P	151	-	P	P	P	P	P	P
		Lot 2								170	-	P	P	P	P	P	P
		Lot 3								219	-	P	P	P	P	P	P
6	滋养透亮补水护手霜	Lot 1	178	P	P	P	P	P	P	251	-	P	P	P	P	P	P
		Lot 2								150	-	P	P	P	P	P	P
		Lot 3								189	-	P	P	P	P	P	P
7	滋润精华磨砂膏	Lot 1	226	P	P	P	P	P	P	193	-	P	P	P	P	P	P
		Lot 2								246	-	P	P	P	P	P	P
		Lot 3								271	-	P	P	P	P	P	P
8	玻尿酸保湿身体乳	Lot 1	226	P	P	P	P	P	P	131	-	P	P	P	P	P	P
		Lot 2								258	-	P	P	P	P	P	P
		Lot 3								106	-	P	P	P	P	P	P
9	滋养焕亮身体乳	Lot 1	226	P	P	P	P	P	P	182	-	P	P	P	P	P	P
		Lot 2								289	-	P	P	P	P	P	P
		Lot 3								224	-	P	P	P	P	P	P
10	滋养修护润发精华素	Lot 1	162	P	P	P	P	P	P	200	-	P	P	P	P	P	P
		Lot 2								143	-	P	P	P	P	P	P
		Lot 3								141	-	P	P	P	P	P	P
11	修护型润唇膏	Lot 1	201	P	P	P	P	P	P	144	-	P	P	P	P	P	P
		Lot 2								144	-	P	P	P	P	P	P
		Lot 3								154	-	P	P	P	P	P	P
12	流光焕采润肤乳	Lot 1	162	P	P	P	P	P	P	162	-	P	P	P	P	P	P
		Lot 2								208	-	P	P	P	P	P	P
		Lot 3								164	-	P	P	P	P	P	P
13	氨基酸洗发露	Lot 1	201	P	P	P	P	P	P	253	-	P	P	P	P	P	P
		Lot 2								112	-	P	P	P	P	P	P
		Lot 3								126	-	P	P	P	P	P	P
14	头皮护理洗发露	Lot 1	162	P	P	P	P	P	P	129	-	P	P	P	P	N	P
		Lot 2								166	-	P	P	P	P	N	P
		Lot 3								180	-	P	P	P	P	N	P
15	婴幼儿沐浴泡泡	Lot 1	167	P	P	P	P	P	P	122	-	P	P	P	P	P	P
		Lot 2								122	-	P	P	P	P	P	P
		Lot 3								189	-	P	P	P	P	P	P
16	剃须泡	Lot 1	162	P	P	P	P	P	P	149	-	P	P	P	P	P	P
		Lot 2								154	-	P	P	P	P	P	P
		Lot 3								217	-	P	P	P	P	P	P
17	净澈去油洗发露	Lot 1	167	P	P	P	P	P	P	101	-	P	P	P	P	P	P
		Lot 2								128	-	P	P	P	P	P	P
		Lot 3								119	-	P	P	P	P	P	P
18	排浊头皮洗发露	Lot 1	201	P	P	P	P	P	P	142	-	P	P	P	P	P	P
		Lot 2								174	-	P	P	P	P	P	P
		Lot 3								239	-	P	P	P	P	P	P
19	香氛沐浴露	Lot 1	167	P	P	P	P	P	P	123	-	P	P	P	P	P	P
		Lot 2								114	-	P	P	P	P	P	P
		Lot 3								119	-	P	P	P	P	P	P
20	多效牙膏	Lot 1	155	P	P	P	P	P	P	178	-	P	P	P	P	P	P
		Lot 2								183	-	P	P	P	P	P	P
		Lot 3								192	-	P	P	P	P	P	P
21	纯白清香沐浴露	Lot 1	155	P	P	P	P	P	P	187	-	P	P	P	P	P	P
		Lot 2								169	-	P	P	P	P	P	P
		Lot 3								273	-	P	P	P	P	P	P
22	莹亮润发精华素	Lot 1	185	P	P	P	P	P	P	121	-	P	P	P	P	P	P
		Lot 2								117	-	P	P	P	P	P	P
		Lot 3								135	-	P	P	P	P	P	P
23	多效修护防晒霜	Lot 1	185	P	P	P	P	P	P	106	-	P	P	P	P	P	P
		Lot 2								206	-	P	P	P	P	P	P
		Lot 3								190	-	P	P	P	P	P	P
24	丝滑轻盈护发素	Lot 1	145	P	P	P	P	P	P	187	-	P	P	P	P	P	P
		Lot 2								207	-	P	P	P	P	P	P
		Lot 3								131	-	P	P	P	P	P	P
25	淡纹抗皱眼霜	Lot 1	145	P	P	P	P	P	P	242	-	P	P	P	P	P	P
		Lot 2								276	-	P	P	P	P	P	P
		Lot 3								252	-	P	P	P	P	P	P
26	肌底精华露	Lot 1	112	P	P	P	P	P	P	272	-	P	P	P	P	P	P
		Lot 2								203	-	P	P	P	P	P	P
		Lot 3								222	-	P	P	P	P	P	P
27	深层洁面乳	Lot 1	145	P	P	P	P	P	P	282	-	P	P	P	P	P	P
		Lot 2								156	-	P	P	P	P	P	P
		Lot 3								134	-	P	P	P	P	P	P
28	美白润肤霜	Lot 1	180	P	P	P	P	P	P	146	-	P	P	P	P	P	P
		Lot 2								162	-	P	P	P	P	P	P
		Lot 3								168	-	P	P	P	P	P	P
29	亮洁皙颜水凝霜	Lot 1	112	P	P	P	P	P	P	118	-	P	P	P	P	P	P
		Lot 2								180	-	P	P	P	P	P	P
		Lot 3								157	-	P	P	P	P	P	P
30	祛斑精华液	Lot 1	180	P	P	P	P	P	P	113	-	P	P	P	P	P	P
		Lot 2								165	-	P	P	P	P	P	P
		Lot 3								165	-	P	P	P	P	P	P
31	奢护面膜	Lot 1	145	P	P	P	P	P	P	142	-	P	P	P	P	P	P
		Lot 2								162	-	P	P	P	P	P	P
		Lot 3								247	-	P	P	P	P	P	P
32	清润黑面膜	Lot 1	185	P	P	P	P	P	P	143	-	P	P	P	P	P	P
		Lot 2								158	-	P	P	P	P	P	P
		Lot 3								163	-	P	P	P	P	P	P
33	角质清洁面膜	Lot 1	185	P	P	P	P	P	P	210	-	P	P	P	P	P	P
		Lot 2								141	-	P	P	P	P	P	P
		Lot 3								295	-	P	P	P	P	P	P
34	清洁面膜	Lot 1	185	P	P	P	P	P	P	159	-	P	P	P	P	P	P
		Lot 2								191	-	P	P	P	P	P	P
		Lot 3								177	-	P	P	P	P	P	P
35	卸妆膏	Lot 1	185	P	P	P	P	P	P	326	-	P	P	P	P	P	P
		Lot 2								246	-	P	P	P	P	P	P
		Lot 3								187	-	P	P	P	P	P	P
36	精华定妆喷雾	Lot 1	185	P	P	P	P	P	P	156	-	P	P	P	P	P	P
		Lot 2								137	-	P	P	P	P	P	P
		Lot 3								135	-	P	P	P	P	P	P
37	维生素乳液	Lot 1	158	P	P	P	P	P	P	215	-	P	P	P	P	P	P
		Lot 2								174	-	P	P	P	P	P	P
		Lot 3								204	-	P	P	P	P	P	P
38	保湿滋润乳	Lot 1	127	P	P	P	P	P	P	125	-	P	P	P	P	P	P
		Lot 2								118	-	P	P	P	P	P	P
		Lot 3								197	-	P	P	P	P	P	P
39	美容洗面奶	Lot 1	158	P	P	P	P	P	P	263	-	P	P	P	P	P	P
		Lot 2								148	-	P	P	P	P	P	P
		Lot 3								170	-	P	P	P	P	P	P
40	保湿洗面奶	Lot 1	127	P	P	P	P	P	P	116	-	P	P	N	P	P	P
		Lot 2								177	-	P	P	N	P	P	P
		Lot 3								165	-	P	P	N	P	P	P
41	保湿舒缓修护乳	Lot 1	127	P	P	P	P	P	P	239	-	P	P	P	P	P	P
		Lot 2								171	-	P	P	P	P	P	P
		Lot 3								164	-	P	P	P	P	P	P
42	水感多效防晒露	Lot 1	158	P	P	P	P	P	P	174	-	P	P	P	P	P	P
		Lot 2								178	-	P	P	P	P	P	P
		Lot 3								154	-	P	P	P	P	P	P
43	抗皱修护晚霜	Lot 1	127	P	P	P	P	P	P	159	-	P	P	P	P	P	P
		Lot 2								162	-	P	P	P	P	P	P
		Lot 3								201	-	P	P	P	P	P	P
44	泡泡洗手液	Lot 1	120	P	P	P	P	P	P	104	-	P	P	N	P	N	P
		Lot 2								110	-	P	P	N	P	N	P
		Lot 3								124	-	P	P	N	P	N	P
45	宝宝爽身粉	Lot 1	120	P	P	P	P	P	P	163	-	P	P	P	P	P	P
		Lot 2								107	-	P	P	P	P	P	P
		Lot 3								148	-	P	P	P	P	P	P

Tab. 3

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类别	功能	功效宣称	作用部位	产品剂型	使用方法	产品名称
普通化妆品	清洁类	清洁	面部	膏霜乳	淋洗	深层洁面乳
						美容洗面奶
						保湿洗面奶
				泥	驻留	氨基酸泥浆精华面膜
				泥	驻留	清洁面膜
			全身皮肤	液体	淋洗	香氛沐浴露
						经典净护沐浴露
						滋养美肤沐浴乳
						爽肤香氛沐浴乳
						婴幼儿沐浴泡泡
				膏霜乳		滋润精华磨砂膏
			头发	液体	淋洗	排浊头皮洗发露
						净澈去油洗发露
						氨基酸洗发露
			手部			泡泡洗手液
		清洁、去屑	头发			头皮护理洗发露
		辅助剃须剃毛	面部			剃须泡
		去角质	面部	膏霜乳	驻留	角质清洁面膜
		卸妆	面部	膏霜乳	驻留	卸妆膏
	护理类	滋养	面部	贴、膜、含基材	驻留	奢护面膜
				贴、膜、含基材		清润黑面膜
				液体		肌底精华露
				膏霜乳		保湿滋润乳
						倍护润肤乳
						维生素乳液
						亮洁皙颜水凝霜
			头发	膏霜乳	淋洗	润发精华素
						丝滑轻盈护发素
						滋养修护润发精华素
						头皮护理润发精华素
			全身皮肤	膏霜乳	驻留	水凝保湿霜
						流光焕采润肤乳
						滋养焕亮身体乳
			手部			保湿倍润护手霜
			手部			舒缓柔嫩恒润护手霜
		爽身	全身皮肤	粉剂	驻留	宝宝清凉粉
		抗皱	眼部	膏霜乳	驻留	淡纹抗皱眼霜
		修护	面部	膏霜乳	驻留	抗皱修护晚霜
			面部		驻留	保湿舒缓修护乳
			头发		淋洗	密集修护发膜
			全身皮肤		驻留	精华修护身体乳
			全身皮肤			玻尿酸保湿身体乳
			口唇	蜡基		修护型润唇膏
	美容修饰类	美容修饰	面部	粉剂	驻留	修容蜜粉
				喷雾剂		精华定妆喷雾
				块状		修容粉饼
特殊化妆品	特殊用途类	防晒	全身皮肤	膏霜乳	驻留	多效修护防晒霜
		防晒	全身皮肤			水感多效防晒露
		美白	全身皮肤			美白润肤霜
		祛斑	面部	液体		祛斑精华液
牙膏	牙膏	—	—	膏霜乳	淋洗	多效牙膏

Suitability study of amplified ATP bioluminescence assay for microbial detection of cosmetics based on two cultivation systems

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