高效液相色谱-串联质谱法测定化妆品中3种甲基咪唑类化合物

doi:10.3724/SP.J.1123.2023.11011

摘要/Abstract

摘要：

建立了一种同时测定化妆品中1-甲基咪唑(1-MEI)、2-甲基咪唑(2-MEI)和4-甲基咪唑(4-MEI)的高效液相色谱-串联质谱(HPLC-MS/MS)方法。样品经乙腈提取、混合型阳离子交换(MCX)固相萃取柱净化、氮吹及复溶后,使用XBridge^® shield RP₁₈色谱柱(150 mm×4.6 mm, 3.5 μm)进行分离;以含0.1%甲酸的20 mmol/L甲酸铵水溶液-乙腈(98∶2, v/v)作为流动相进行等度洗脱;质谱分析采用电喷雾电离(ESI)源和多反应监测(MRM)模式,外标法定量。结果表明,1-MEI在5~200 μg/L范围内线性关系良好(相关系数(r²)≥0.9994), 2-MEI和4-MEI在2~100 μg/L范围内线性关系良好(r²≥0.9984)。3种甲基咪唑类化合物的检出限(LOD)为10~30 μg/kg,定量限(LOQ)为25~100 μg/kg。在低、中、高3个加标水平下,不同空白基质中3种甲基咪唑类化合物的回收率为80.9%~107.9%,相对标准偏差(RSD, n=6)为1.2%~12.8%。将该方法应用于48个化妆品样品的检测,结果表明,有9个样品检出4-MEI,检出含量为26~1000 μg/kg; 2个样品检出2-MEI,检出含量分别为240 μg/kg和267 μg/kg;而48个样品中均未检出1-MEI。该方法简单、快速,灵敏度高,为化妆品中3种甲基咪唑类化合物的风险评估和筛查监测提供了技术支撑。

关键词: 高效液相色谱-串联质谱, 1-甲基咪唑, 2-甲基咪唑, 4-甲基咪唑, 化妆品

Abstract:

Methylimidazole compounds are byproducts formed during the caramel-coloring process and are used in various cosmetics. In addition metronidazole is an antibacterial and anti-inflammatory drug commonly used in modern medicine and is used in cosmetics to treat acne in the short-term. The illegal addition of metronidazole during cosmetics production can result in residual 2-methylimidazole (2-MEI), which, along with 4-methylimidazole (4-MEI), is a class 2B carcinogen. Therefore, establishing efficient, accurate, and sensitive analytical techniques for analyzing methylimidazole compounds in cosmetics is an urgent objective. In this study, a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for simultaneously determining 1-methylimidazole (1-MEI), 2-MEI, and 4-MEI in cosmetics was developed. Cosmetics samples were extracted via ultrasonication in acetonitrile and purified using a mixed cation-exchange (MCX) solid-phase extraction (SPE) column, with subsequent drying under a stream of nitrogen and redissolution in acetonitrile. The resulting solution was then filtered through a 0.22 μm organic filter membrane for further testing. The analytes were separated using an XBridge^® shield RP₁₈ chromatographic column (150 mm×4.6 mm, 3.5 μm) and isocratically eluted with 20 mmol/L ammonium formate solution (containing 0.1% formic acid)-acetonitrile (98∶2, v/v). The target compounds were ionized by electrospray ionization (ESI) source, analyzed in multi-reaction monitoring (MRM) mode, and quantified using the external standard method, with the peak area of the quantitative ion and the mass concentration of the compound taken as the longitudinal and transverse coordinates, respectively. Matrix-matching working curves were also constructed. 1-MEI exhibited good linear relationships in the range of 5-200 μg/L, with correlation coefficients (r²)≥0.9994, while 2-MEI and 4-MEI showed good linearities in the range of 2-100 μg/L with r²≥0.9984. The three methylimidazole compounds exhibited limits of detection (LODs) and quantification (LOQs) of 10-30 μg/kg and 25-100 μg/kg, respectively. Under three spiked levels (LOQ, 2LOQ, 10LOQ), the recoveries of three methylimidazole compounds were 80.9%-107.9%, with relative standard deviations (RSDs, n=6) of 1.2%-12.8%. The practicability of the method was examined using 48 cosmetic samples; 4-MEI was detected in nine samples at contents of 26-1000 μg/kg, while two samples contained 240 and 267 μg/kg of 2-MEI, respectively. 1-MEI was not detected in any of the 48 samples tested. The developed method is simple, fast, and highly sensitive, and provides methodological support for assessing risks and monitoring the three methylimidazole compounds in cosmetics through screening.

Key words: high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), 1-methylimidazole (1-MEI), 2-methylimidazole (2-MEI), 4-methylimidazole (4-MEI), cosmetics

中图分类号:

O658

蔡翠玲, 李菊, 席静, 侯颖烨, 曾广丰, 谢建军, 吴舒悦, 王璐. 高效液相色谱-串联质谱法测定化妆品中3种甲基咪唑类化合物[J]. 色谱, 2024, 42(11): 1052-1058.

CAI Cuiling, LI Ju, XI Jing, HOU Yingye, ZENG Guangfeng, XIE Jianjun, WU Shuyue, WANG Lu. Determination of three methylimidazole compounds in cosmetics by high performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2024, 42(11): 1052-1058.

图/表 5

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Compound	Retention time/min	Precursor ion (m/z)	Product ion (m/z)	DP/V	CE/eV
1-Methylimidazole (1-MEI)	2.71	83.2	56.1^*	55	26.9
			42.2	55	39.9
2-Methylimidazole (2-MEI)	2.85	83.2	42.2^*	55	39.9
			56.1	55	26.9
4-Methylimidazole (4-MEI)	3.05	83.2	56.1^*	55	26.9
			42.2	55	39.9

Compound	Retention time/min	Precursor ion (m/z)	Product ion (m/z)	DP/V	CE/eV
1-Methylimidazole (1-MEI)	2.71	83.2	56.1^*	55	26.9
			42.2	55	39.9
2-Methylimidazole (2-MEI)	2.85	83.2	42.2^*	55	39.9
			56.1	55	26.9
4-Methylimidazole (4-MEI)	3.05	83.2	56.1^*	55	26.9
			42.2	55	39.9

Compound	Linear range/ (μg/L)	Toner matrix		Volatile oil matrix		Cream matrix		Loose powder matrix		LOD/ (μg/kg)	LOQ/ (μg/kg)
Compound	Linear range/ (μg/L)	Linear equation	r²	Linear equation	r²	Linear equation	r²	Linear equation	r²	LOD/ (μg/kg)	LOQ/ (μg/kg)
1-MEI	5-200	Y=254X+9	0.9996	Y=264X+307	0.9994	Y=235X+1320	0.9996	Y=358X+3210	0.9998	30	100
2-MEI	2-100	Y=1140X-284	0.9996	Y=1320X+1570	0.9994	Y=1180X+2420	0.9990	Y=1310X+2200	0.9984	10	25
4-MEI	2-100	Y=3210X+5620	0.9998	Y=3240X+9870	0.9991	Y=3260X+8480	0.9998	Y=2370X+30000	0.9996	10	25

Compound	Linear range/ (μg/L)	Toner matrix		Volatile oil matrix		Cream matrix		Loose powder matrix		LOD/ (μg/kg)	LOQ/ (μg/kg)
Compound	Linear range/ (μg/L)	Linear equation	r²	Linear equation	r²	Linear equation	r²	Linear equation	r²	LOD/ (μg/kg)	LOQ/ (μg/kg)
1-MEI	5-200	Y=254X+9	0.9996	Y=264X+307	0.9994	Y=235X+1320	0.9996	Y=358X+3210	0.9998	30	100
2-MEI	2-100	Y=1140X-284	0.9996	Y=1320X+1570	0.9994	Y=1180X+2420	0.9990	Y=1310X+2200	0.9984	10	25
4-MEI	2-100	Y=3210X+5620	0.9998	Y=3240X+9870	0.9991	Y=3260X+8480	0.9998	Y=2370X+30000	0.9996	10	25

Compound	Spiked level/ (μg/kg)	Toner matrix		Volatile oil matrix		Cream matrix		Loose powder matrix
Compound	Spiked level/ (μg/kg)	Recovery/%	RSD/%	Recovery/%	RSD/%	Recovery/%	RSD/%	Recovery/%	RSD/%
1-MEI	100	90.8	5.5	84.0	3.3	89.7	3.7	83.0	2.7
	200	91.3	10.1	85.4	5.3	84.4	3.2	83.1	3.6
	1000	82.0	1.2	83.8	2.2	83.7	2.2	80.9	1.9
2-MEI	25	107.9	8.8	87.9	9.4	93.0	5.0	90.2	5.0
	50	104.7	4.2	91.7	5.8	101.7	11.3	84.7	3.8
	250	87.8	6.6	88.6	3.4	85.6	6.0	84.8	2.3
4-MEI	25	94.4	12.2	93.8	12.8	85.5	5.3	85.5	5.6
	50	89.3	2.2	100.8	4.2	101.4	9.6	89.4	7.2
	250	86.4	6.7	89.5	5.7	82.8	2.8	84.9	2.3