Determination of new glucocorticoid called clobetasol acetate in cosmetics by ultra performance liquid chromatography-tandem mass spectrometry

doi:10.3724/SP.J.1123.2022.06010

Abstract

Abstract:

At present, new prohibited substances are becoming more common illegal additions in cosmetics. Clobetasol acetate is a new glucocorticoid, which is not covered in the current national standards and is a homologue of clobetasol propionate. A method was established for the determination of clobetasol acetate as a new glucocorticoid (GC) in cosmetics by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Five common cosmetic matrices were suitable for this new method: creams, gels, clay masks, masks and lotions. Four pretreatment methods were compared: direct extraction by acetonitrile, PRiME pass-through column purification, solid-phase extraction (SPE) purification, and QuEChERS purification. Further, the effects of different extraction efficiencies of the target compound, such as extraction solvents and extraction time, were investigated. The MS parameters, such as ion mode, cone voltage and collision energy of ion pairs of the target compound, were optimized. The chromatographic separation conditions and response intensities of the target compound in different mobile phases were compared. Based on the experimental results, the optimal extraction method was determined to be direct extraction, wherein the samples were vortex dispersed with acetonitrile, ultrasonic extraction over 30 min and filtered by a 0.22 μm organic millipore filter, and then the samples were detected by UPLC-MS/MS. The concentrated extracts were separated on a Waters CORTECS C₁₈ column (150 mm×2.1 mm, 2.7 μm), where the water and acetonitrile were used as the mobile phases for gradient elution. The target compound was detected with the multiple reaction monitoring (MRM) mode under electrospray ionization and positive ion scanning (ESI⁺). Quantitative analysis was performed by matrix matching standard curve. Under the optimum conditions, the target compound had good linear fitting in the range of 0.9-37 μg/L. The linear correlation coefficient (R²) was greater than 0.99, the limit of quantification (LOQ) of the method was 0.09 μg/g and the limit of detection (LOD) was 0.03 μg/g for these five different cosmetic matrices. The recovery test was conducted under three spiked levels: 1, 2 and 10 times of LOQ. The recoveries of the tested substance were between 83.2% and 103.2% in these five cosmetic matrices, and the relative standard deviations (RSDs, n=6) were between 1.4% and 5.6%. This method was used to screen cosmetic samples of different matrix types, and a total of five positive samples were found, in which the content range of clobetasol acetate was from 1.1 to 48.1 μg/g. In conclusion, the method is simple, sensitive and reliable, and is suitable for high-throughput qualitative and quantitative screening, and the analysis of cosmetics with different matrix types. Moreover, the method provides crucial technical support and a theoretical basis for the establishment of feasible detection standards for clobetasol acetate in China, as well as for the control of the compound in cosmetics. This method has important practical significance to implement management measures of illegal additions in cosmetics.

Key words: ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), glucocorticoids, clobetasol acetate, cosmetics

CLC Number:

O658

YANG Piaopiao, HUANG Wei, LI Lixia, LIU Hong. Determination of new glucocorticoid called clobetasol acetate in cosmetics by ultra performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2023, 41(3): 250-256.

Figures/Tables 9

Fig. 1 Structural formula of clobetasol acetate

Table 1 Retention time and MS parameters for clobetasol acetate

Compound	Retention time/ min	Parent ion (m/z)	Quantitative/ qualitative ions (m/z)	Cone voltage/ V	Collison energies/ V
Clobetasol	4.67	453.2	278.2/263.2	24	24/28
acetate

Fig. 2 Extraction recoveries of the target compound under different sample pretreatment methods (n=6) ①: extraction by acetonitrile; ②: solid-phase extraction; ③: PRiME pass-through column purification; ④: QuEChERS purification.

Fig. 3 Extraction recoveries of the target compound under different extraction solvents (n=6) ①: acetonitrile; ②: acetonitrile-water (70∶30, v/v); ③: acetonitrile-water (50∶50, v/v); ④: methanol; ⑤: methanol-water (70∶30, v/v); ⑥: methanol-water (50∶50, v/v).

Fig. 4 Extraction recoveries of the target compound under different ultrasonic times (n=6)

Table 2 Linear ranges, linear equations, correlation coefficients (R2), limits of detection (LODs), limits of quantification (LOQs), and matrix effects (MEs) of the target compound under different matrix types

Matrix type	Linear range/(μg/L)	Linear equation	R²	LOD/(μg/g)	LOQ/(μg/g)	ME/%
Standard solution	0.9-37	y=2.57×10³x+6.39×10²	0.9998	0.03	0.09	/
Cream	0.9-37	y=1.34×10³x+5.04×10²	0.9982	0.03	0.09	52.2
Gel	0.9-37	y=8.73×10²x+2.17×10²	0.9958	0.03	0.09	34.0
Clay masks	0.9-37	y=1.89×10³x+9.13×10²	0.9984	0.03	0.09	73.7
Mask	0.9-37	y=1.95×10³x+9.02×10²	0.9998	0.03	0.09	76.0
Lotion	0.9-37	y=2.15×10³x+1.08×10³	0.9994	0.03	0.09	83.8

Table 3 Spiked recoveries and precisions of the target compound in different matrix types (n=6)

Compound	Matrix type	0.09 μg/g		0.19 μg/g		0.94 μg/g
Compound	Matrix type	Recovery/%	RSD/%	Recovery/%	RSD/%	Recovery/%	RSD/%
Clobetasol	cream	94.4	3.3	96.2	2.2	101.1	5.6
acetate	gel	85.5	2.4	100.4	3.8	95.6	4.4
	clay mask	94.0	5.3	91.9	4.4	92.4	1.4
	mask	91.3	3.8	100.1	3.9	103.2	3.6
	lotion	83.2	3.9	94.8	3.2	96.5	1.4

Table 4 Information and contents of positive samples

No.	Matrix type	Manufacturer	Sampling site	Content/ (μg/g)
1	mask	manufacturer 1	small shop	1.1
2	mask	manufacturer 1	small shop	1.1
3	mask	manufacturer 2	small shop	34.5
4	gel	manufacturer 3	online shopping	39.2
5	gel	manufacturer 3	production unit	48.1

Fig. 5 Chromatograms of clobetasol acetate in (a) typical positive and (b)reference solution

References

[1]	Yang B F, Chen J G. Pharmacology. 9th ed. Beijing: People’s Medical Publishing House, 2018
[1]	杨宝峰, 陈建国. 药理学. 9版. 北京: 人民卫生出版社, 2018
[2]	You Q D. Medicinal Chemistry. 8th ed. Beijing: People’s Medical Publishing House, 2016
[2]	尤启东. 药物化学. 8版. 北京: 人民卫生出版社, 2016
[3]	Yang P P, Liu H, Li L X. China Surfactant Detergent & Cosmetics, 2021, 51(5): 468
[3]	杨飘飘, 刘红, 李丽霞. 日用化学工业, 2021, 51(5): 468
[4]	Luo H T, Huang X L, Wu H Q, et al. Chinese Journal of Analytical Chemistry, 2017, 45(9): 1381
[4]	罗辉泰, 黄晓兰, 吴惠勤, 等. 分析化学, 2017, 45(9): 1381
[5]	Qiao Y S, Dong Y L, Huang C F, et al. China Surfactant Detergent & Cosmetics, 2021, 51(12): 1259
[5]	乔亚森, 董亚蕾, 黄传峰, 等. 日用化学工业, 2021, 51(12): 1259
[6]	China Food and Drug Administration. Safety and Technical Standards for Cosmetics (2015 Edition).(2015-12-23) [2022-03-14]. https://www.nmpa.gov.cn/directory/web/nmpa/images/MjAxNcTqtdoyNji6xbmruOa4vbz+LnBkZg==.pdf
[6]	国家食品药品监督管理局. 化妆品安全技术规范(2015年版).(2015-12-23) [2022-03-14]. https://www.nmpa.gov.cn/directory/web/nmpa/images/MjAxNcTqtdoyNji6xbmruOa4vbz+LnBkZg==.pdf
[7]	Korea Cosmetics Regulation. General Administration of Quality Supervision, Inspection and Quarantine of the People’s Republic of China, transl. Beijing: China Metrology Press, 2013
[7]	韩国化妆品法规. 中华人民共和国国家质量监督检验检疫总局, 译. 北京: 中国计量出版社, 2013
[8]	Japan Cosmetics Regulation. General Administration of Quality Supervision, Inspection and Quarantine of the People’s Republic of China, transl. Beijing: China Metrology Press, 2011
[8]	日本化妆品法规. 中华人民共和国国家质量监督检验检疫总局, 译. 北京: 中国计量出版社, 2011
[9]	EC) No 1223/2009
[10]	Association of Southeast Asian Nations ASEAN Cosmetics Regulations. General Administration of Quality Supervision, Inspection and Quarantine of the People’s Republic of China, transl. Beijing: China Metrology Press, 2010
[10]	东盟化妆品法规. 中华人民共和国国家质量监督检验检疫总局, 译. 北京: 中国计量出版社, 2010
[11]	GB/T 24800.2-2009
[12]	GB/T 40145-2021
[13]	SN/T 2533-2010
[14]	SN/T 4504-2016
[15]	Li Y J, Zhao X L, Cen L Y, et al. Flavour Fragrance Cosmetics, 2018(1): 58
[15]	李扬杰, 赵祥龙, 岑立怡, 等. 香精香料化妆品, 2018(1): 58
[16]	Luo H T, Huang X L, Wu H Q, et al. Chinese Journal of Chromatography, 2017, 35(8): 816
[16]	罗辉泰, 黄晓兰, 吴惠勤, 等. 色谱, 2017, 35(8): 816
[17]	Liu H, Yang P P, Li L X. Journal of Instrumental Analysis, 2020, 39(9): 1112
[17]	刘红, 杨飘飘, 李丽霞. 分析测试学报, 2020, 39(9): 1112
[18]	Liu H, Zhang J Y, Hu B, et al. Flavour Fragrance Cosmetics, 2020(4): 83
[18]	刘红, 张静雅, 胡贝, 等. 香料香精化妆品, 2020(4): 83
[19]	Weng D H, Liu J, Jiang Q Q, et al. Flavour Fragrance Cosmetics, 2019(3): 45
[19]	翁东海, 刘江, 蒋清清, 等. 香料香精化妆品, 2019(3): 45
[20]	Pan X H, Yin S, Liu Y L, et al. Chinese Journal of Chromatography, 2018, 36(4): 356
[20]	潘小红, 殷帅, 刘玉玲, 等. 色谱, 2018, 36(4): 356
[21]	Dong Y L, Liu Z, Wang H Y, et al. China Surfactant Detergent & Cosmetics, 2020, 50(12): 885
[21]	董亚蕾, 刘钊, 王海燕, 等. 日用化学工业, 2020, 50(12): 885
[22]	Yang G Y, Sha L N, Liang Q Y, et al. Chinese Journal of Analysis Laboratory, 2021, 40(8): 959
[22]	杨光勇, 沙丽娜, 梁秋艳, 等. 分析试验室, 2021, 40(8): 959
[23]	Zhangzhou Health Commission. Notification on the Progress of Investigation and Disposal of “Ouai Antibacterial Cream” Incident. (2021-01-17) [2022-03-10]. http://wjw.zhangzhou.gov.cn/cms/html/zzswshjhsywyh/2021-01-17/1663659826.html
[23]	漳州市卫生健康委员会. 关于“欧艾抑菌霜”事件调查处置进展情况的通报. (2021-01-17) [2022-03-10]. http://wjw.zhangzhou.gov.cn/cms/html/zzswshjhsywyh/2021-01-17/1663659826.html
[24]	Meng X S, Bai H, Guo T, et al. J Chromatogr A, 2017, 1528: 61
[25]	Jian L H, Yuan X Q, Han J, et al. Rapid Commun Mass Spectrom, 2021, 35(3): e8999
[26]	Zhan J, Ni M L, Zhao H Y, et al. J Sep Sci, 2014, 37(24): 3684
[27]	Jelena B, GoluboviĆ B M O A, ZeĆeviĆ M L. J Sep Sci, 2014, 37(24): 3684
[28]	Jessica F, Vincenza A. J Pharm Biomed Anal, 2014, 91: 185
[29]	He M L, Guo C C, Shi F, et al. Chinese Journal of Pharmaceutical Analysis, 2019, 39(1): 105
[29]	贺美莲, 郭常川, 石峰, 等. 药物分析杂志, 2019, 39(1): 105
[30]	Wang B F, Xu L, Xu Z Z, et al. Food Science, 2021, 42(7): 301
[30]	王冰峰, 徐雷, 徐贞贞, 等. 食品科学, 2021, 42(7): 301
[31]	Xu D M, Zeng S M, Liu X C, et al. Chinese Journal of Chromatography, 2022, 40(5): 423
[31]	徐敦明, 曾三妹, 柳训才, 等. 色谱, 2022, 40(5): 423
[32]	Zhou Y S, Gong J, Yang K X, et al. Chinese Journal of Chromatography, 2022, 40(2): 165
[32]	周永顺, 龚剑, 杨可欣, 等. 色谱, 2022, 40(2): 165
[33]	Yao Q, Guo Y, Xue J, et al. J Pharm Biomed Anal, 2020, 179: 112980
[34]	Liu M, Li X, Li J, et al. J Colloid Interface Sci, 2017, 504: 124
[35]	Giaccone V, Macaluso A, Cammilleri G, et al. Food Add Contam B, 2018, 11(3): 201