高效液相色谱法同时测定淋洗类化妆品中7种异噻唑啉酮类防腐剂及质谱确证

doi:10.3969/j.issn.2097-2806.2024.10.015

摘要/Abstract

摘要：

建立了同时检测淋洗类化妆品中7种异噻唑啉酮类防腐剂的高效液相色谱分析方法及液相色谱-串联质谱确证方法。膏霜乳和液体两种不同基质类型的化妆品样品分别经甲醇超声提取，提取液以离心处理后，取上清液经微孔滤膜过滤后测定。经CAPCELL PAK C₁₈ MGIII （4.6 mm×250 mm, 5 μm）色谱柱分离，以甲醇和0.1%磷酸水溶液为流动相梯度洗脱，流速1.0 ml/min，柱温40 ℃。检测波长为275，282，和319 nm。以高效液相色谱法的结果进行定性筛查，并采用标准曲线法定量，以液相色谱-串联质谱法的结果进行确证。结果表明，7种异噻唑啉酮类防腐剂在0.1~50 mg/L质量浓度范围内线性关系良好，相关系数均≥0.999 9。在低、中、高3种加标浓度水平下，平均加标回收率分别为97.5%~109.9% （膏霜乳）和95.6%~103.6% （液体）；相对标准偏差（RSD）小于3.0%，检出限为0.05~0.11 mg/kg。该方法具有简单、快速、准确性好的特点，适用于淋洗类化妆品中7种异噻唑啉酮类防腐剂的含量测定及质谱确证。

关键词: 淋洗类化妆品, 高效液相色谱法, 质谱确证, 异噻唑啉酮类防腐剂

Abstract:

An analytical method for the simultaneous determination of seven isothiazolinone preservatives in rinse-off cosmetics by high-performance liquid chromatography （HPLC） and verification by liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） was established. The cream and liquid cosmetic samples were ultrasonically extracted with methanol, centrifuged, and the supernatant was filtered through a microporous membrane for determination. The separation was executed on a CAPCELL PAK C₁₈ MGIII column by gradient elution at a flow rate of 1.0 mL/min with methanol and 0.1% （V/V） phosphoric acid aqueous solution as the mobile phases. The column temperature was set at 40 ℃. The wavelengths of detection were 278, 282 and 319 nm. The results of high performance liquid chromatography were used for qualitative screening, the standard curve method was used for quantification, and the results of liquid chromatography tandem mass spectrometry were used for verification. The results show that the linear ranges of the seven isothiazolinone preservatives are 0.1-50 mg/L, and the correlation coefficients are all greater than 0.999 9. The average recoveries are 97.5%-109.9% （for cream） and 95.6%-103.6% （for liquid）, and the relative standard deviations （RSD） are bellow 3.0% at low, medium and high spiked levels. The detection limits are 0.05-0.11 mg/kg. The method is simple, rapid and accurate for the simultaneous determination and verification of seven isothiazolinone preservatives in the rinse-off cosmetics.

Key words: rinse-off cosmetics, high performance liquid chromatography, mass spectrometric verification, isothiazolinone preservatives

中图分类号:

TQ658

陈岑, 崔邶周, 张丽媛, 颜琳琦, 程巧鸳. 高效液相色谱法同时测定淋洗类化妆品中7种异噻唑啉酮类防腐剂及质谱确证[J]. 日用化学工业（中英文）, 2024, 54(10): 1268-1275.

Cen Chen, Beizhou Cui, Liyuan Zhang, Linqi Yan, Qiaoyuan Cheng. Simultaneous determination of seven isothiazolinone preservatives in rinse-off cosmetics by HPLC and verification by HPLC-MS/MS[J]. China Surfactant Detergent & Cosmetics, 2024, 54(10): 1268-1275.

图/表 9

表1

图1

图2

图3

表2

表3

表4

图4

图5

参考文献 20

[1]	Fu Zhongye, Wang Qing, Jia Lanni, et al. The synthesis and application of isothiazolone derivatives for recent decade[J]. Materials Reports, 2012, 26 (S1) : 291-296.
[2]	Aerts O, Goossens A, Julien Lambert. Contact allergy caused by isothiazolinone derivatives: an overview of non-cosmetic and unusual cosmetic sources[J]. European Journal of Dermatology, 2017, 27 (2) : 115-122.
[3]	Felmingham C, Nixon R, Palmer A, et al. Allergic contact dermatitis caused by benzisothiazolinone in a continuous positive airway pressure mask liquid soap[J]. Contact Dermatitis, 2019, 82 (2) : 152-153.
[4]	Silva V, Silva C, Soares P, et al. Isothiazolinone biocides: chemistry, biological, and toxicity profiles[J]. Molecules, 2020, 25 (4) : 991-1013.
[5]	Ito M, Mochida K, Ito K, et al. Induction of apoptosis in testis of the marine teleost mummichog Fundulus heteroclitus after in vivo exposure to the antifouling biocide 4, 5-dichloro-2-n-octyl-3 (2H) -isothiazolone (Sea-Nine 211)[J]. Chemosphere, 2013, 90 (3) : 1053-1060.
[6]	Lv Peng, Xu Jiaqing, Wang Sen, et al. Studies on acute toxicity and oxidative stress of benzisothiazolin to danio rerio embryos[J]. Biotechnology Bulletin, 2018, 34 (1) : 172-182. doi: 10.13560/j.cnki.biotech.bull.1985.2017-0665
[7]	European Union. Commission Regulation (EU) 2017/1224 of 6 July 2017 amending Annex V to Regulation(EC)No 1223/2009 of the European Parliament and of the Council on Cosmetic Products[EB/OL]. (2017-07-06)[2023-07-07]. http://data.europa.eu/eli/reg/2017/1224/oj.
[8]	Burnett C L, Boyer I, Bergfeld W F, et al. Amended safety assessment of methylisothiazolinone as used in cosmetics[J]. International Journal of Toxicology, 2019, 38 (S1): 70S-84S.
[9]	Su Zhe, Lin Qingbing, Wang Gangli, et al. A review on the latest development of safety risk assessment technical regulation of cosmetic preservative methylisothiazolinone[J]. Flavour Fragrance Cosmetics, 2019 (1) : 77-82.
[10]	China Food and Drug Administration. Safety and Technical Standards for Cosmetics (2015)[S]. Beijing: China Food and Drug Administration, 2016.
[11]	He Xiaoling, Ma Yan, Deng Xing, et al. Current status of standards and research progress of detection methods of isothiazolinone preservatives[J]. China Surfactant Detergent & Cosmetics, 2023, 53 (10) : 1186-1193.
[12]	Zhang Jianmei, Pan Ya’nan, Bai Hua. Determination of the content of active substances in isothiazolone derivatives by UV spectrophotometry[J]. Industrial Water Treatment, 2011, 31 (5) : 70-71. doi: 10.11894/1005-829x.2011.31.（05）.70
[13]	Lin Junfeng, Wang Chengyun, Bai Shuang, et al. Simultaneous determination of seven restricted isothiazolinone antibacterial agents in textiles[J]. Textile Auxiliaries, 2022, 39 (7) : 58-64.
[14]	Wang Chengyun, Zhu Naiqing, Li Yongtao, et al. Determination of seven restricted isothiazolinone antibacterial agents in leather and leather products by gas chromatography/tandem mass spectrometry[J]. Westleather, 2021, 43 (7) : 22-26.
[15]	Li Guangdong. Determination of five isothiazolinone biocides in waterborne coatings by LC-MS/MS[J]. Paint & Coatings Industry, 2019, 49 (12) : 54-61.
[16]	Wang Jin, Ren Jiefang, Zhang Donglei, et al. Determination of three isothiazolinone preservatives in children’s color clay modeling toys by high performance liquid chromatography-tandem mass spectrometry[J]. Journal of Instrumental Analysis, 2019, 38 (11) : 1384-1388.
[17]	Yang Yongchao, Yan Jing, Zhao Haolibao, et al. Determination of three isothiazolinone preservatives in children products by high performance liquid chromatography-tandem mass spectrometry[J]. Journal of Analytical Science, 2019, 35 (1) : 80-84.
[18]	Wei Zhidong, Han Feng. Determination of isothiazolinones preservatives in house-hold detergents by ultra-performance liquid chromatography-tandem mass spectrometry[J]. Anhui Chemical Industry, 2018, 44 (3) : 105-107.
[19]	Pan Jianting. Detection of five kinds of isothiazolinones preservatives in food contact paper packaging[J]. Yunnan Chemical Technology, 2022, 49 (2) : 37-39.
[20]	National Medical Products Administration. Circular of National Medical Products Administration on Incorporating 7 Inspect ion Methods, including the Inspection Method for Preservatives in Cosmetics, into the Safety and Technical Standards for Cosmetics (2015 Edition)(No. 17, 2021)[EB/OL]. (2021-02-18)[2023-07-07]. https://www.nmpa.gov.cn/xxgk/ggtg/qtggtg/20210302174406138.html.

序号	化合物	母离子（m/z）	子离子（m/z）	CE/eV	相对离子丰度/%	允许相对偏差/%
1	MIT	116.0	100.8^*，84.9	31, 33	100, 50	25
2	CMIT	150.0	87.0^*，115.0	51, 29	100, 22	25
3	BIT	152.0	134.1^*，77.1	39, 33	100, 23	25
4	MBIT	166.0	151.0^*，96.0	33, 48	100, 65	20
5	BBIT	208.0	151.9^*，108.9	24, 34	100, 37	25
6	OIT	214.1	102.1^*，57.2	21, 22	100, 53	20
7	DCOIT	282.0	170.0^*，71.0	23, 26	100, 32	25

化合物	保留时间/min	线性范围/（mg/L）	回归方程	相关系数r	LOD/（mg/kg）	LOQ/（mg/kg）
MIT	3.20	0.1~50	Y=44 505X+3 104	0.999 9	0.05	0.18
CMIT	5.17	0.1~50	Y=27 280X+1 562	1.000 0	0.09	0.31
BIT	6.43	0.1~50	Y=22 017X+661	1.000 0	0.11	0.37
MBIT	7.07	0.1~50	Y=20 914X+800	1.000 0	0.10	0.32
BBIT	9.53	0.1~50	Y=15 594X+976	1.000 0	0.08	0.39
OIT	10.20	0.1~50	Y=23 071X+1 663	0.999 9	0.09	0.27
DCOIT	12.72	0.1~50	Y=19 385X+851	1.000 0	0.08	0.26

化合物	加标浓度水平/（mg/kg）	平均回收率/%		RSD/%
化合物	加标浓度水平/（mg/kg）	膏霜乳	液体	膏霜乳	液体
MIT	5	105.6	99.8	2.50	0.62
	50	104.1	100.2	0.98	0.69
	400	98.6	95.7	1.67	1.42
CMIT	5	105.9	96.6	1.45	1.60
	50	104.3	101.0	0.98	0.86
	400	102.9	99.6	1.67	1.41
BIT	5	109.9	97.6	1.07	1.61
	50	104.1	102.2	0.96	0.70
	400	98.4	95.8	0.97	0.38
MBIT	5	105.7	103.6	2.02	1.95
	50	103.7	102.4	0.92	0.76
	400	97.5	95.6	0.75	0.77
BBIT	5	101.8	97.7	1.73	1.84
	50	105.7	102.1	0.94	0.76
	400	102.7	99.6	1.67	1.38
OIT	5	98.6	95.7	1.64	2.52
	50	102.9	102.5	0.92	0.67
	400	98.8	97.6	0.36	0.71
DCOIT	5	104.1	99.8	1.02	1.83
	50	104.5	101.1	0.99	0.79
	400	101.9	100.3	0.50	0.35

化合物	检出限/ ng	定量限/ ng	检出浓度/（mg/kg）	定量浓度/（mg/kg）
MIT	1.02	3.42	0.001 0	0.003 4
CMIT	1.08	3.61	0.001 1	0.003 6
BIT	1.88	6.25	0.001 9	0.006 3
MBIT	0.51	1.72	0.000 51	0.001 7
BBIT	0.35	1.17	0.000 35	0.001 2
OIT	0.28	0.93	0.000 28	0.000 9
DCOIT	1.64	5.47	0.001 6	0.005 5