化妆品中13种防腐剂的高效液相色谱分析及质谱确证

doi:10.3969/j.issn.2097-2806.2024.12.016

摘要/Abstract

摘要：

建立了同时测定化妆品中13种防腐剂（乙酰丙酸、对羟基苯乙酮、覆盆子酮、对茴香酸、辛酰羟肟酸、羟基乙氧基苯基丁酮、甲基异噻唑啉酮、苯氧乙醇、苯甲酸、4-羟基苯甲酸甲酯、氯苯甘醚、脱氢乙酸、5-溴-5-硝基-1,3-二噁烷）的高效液相色谱分析方法和质谱确证方法。不同基质类型的化妆品样品经甲醇超声提取后，使用Agilent ZORBAX Eclipse XDB-C18 （250 mm×4.6 mm，5 μm）色谱柱进行色谱分离，以0.1%磷酸水溶液和乙腈作为流动相梯度洗脱，流速1.0 mL/min，柱温30 ℃，检测波长为275，230和210 nm。对疑似阳性的样品，进一步采用液相色谱-串联质谱法或气相色谱-质谱法进行确证。结果表明，13种防腐剂组分在各自的质量浓度范围内，线性关系良好（r>0.999 8），检出限为0.4~100.0 mg/kg，定量限为1.2~250.0 mg/kg。在低、中、高3个加标水平下，13种目标组分在3种基质样品中的平均回收率为84.0%~115.4%，相对标准偏差为0.5%~4.8%（n=6）。该方法操作简便、分离效果好、精密度高，适用于化妆品中常见表外防腐剂和准用防腐剂的定性和定量测定。

关键词: 防腐剂, 化妆品, 高效液相色谱法, 质谱

Abstract:

A method using high performance liquid chromatography （HPLC） was developed for the simultaneous determination of 13 preservatives （levulinic acid, p-hydroxyacetophenone, raspberry ketone, p-anisic acid, caprylhydroxamic acid, hydroxyethoxyphenyl butanone, methylisothiazolinone, phenoxyethanol, benzoic acid, methylparaben, chlorphenesin, dehydroacetic acid, and 5-bromo-5-nitro-1,3-dioxane） in cosmetics. Different types of samples were ultrasonically extracted by methanol, then the separation of 13 preservatives was carried out on a column of Agilent ZORBAX Eclipse XDB-C18 （250 mm×4.6 mm, 5 μm） by gradient elution at a flow rate of 1.0 mL/min, using 0.1% phosphoric acid solution and acetonitrile as mobile phases. The column temperature was 30 ℃, and the detection was completed by a diode array detector with the wavelengths at 275, 230 and 210 nm. Suspected positive samples were further confirmed by liquid chromatography-tandem mass spectrometry or gas chromatography-mass spectrometry. The linear regression analysis data shows good linearity for 13 preservatives in the respective mass concentration range, with their correlation coefficients （r） greater than 0.999 8. The limits of detection （LODs） and limits of quantitation （LOQs） of the method are in the ranges of 0.4-100.0 mg/kg and 1.2-250.0 mg/kg, respectively. At three spiked levels, the average recoveries for 13 target compounds in three kinds of matrix samples are within 84.0%-115.4%, and the relative standard deviations （RSD） are within 0.5%-4.8% （n = 6）. This method is convenient, efficient, and precise, which can be used for qualitative and quantitative analysis of common preservatives in daily cosmetics.

Key words: preservatives, cosmetics, high performance liquid chromatography, mass spectrometry

中图分类号:

TQ658

邹坡, 徐炜, 李雪艳, 李长于, 陈艳, 陈勤勤. 化妆品中13种防腐剂的高效液相色谱分析及质谱确证[J]. 日用化学工业（中英文）, 2024, 54(12): 1532-1540.

Po Zou, Wei Xu, Xueyan Li, Changyu Li, Yan Chen, Qinqin Chen. Determination of 13 preservatives in cosmetics by high performance liquid chromatography and verification by mass spectrometry[J]. China Surfactant Detergent & Cosmetics, 2024, 54(12): 1532-1540.

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参考文献 22

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序号	组分名称	ρ（系列混合标准溶液）/（μg/mL）
1	甲基异噻唑啉酮	0.12	0.30	1.20	3.00	12.00	30.00
2	乙酰丙酸	25.0	62.5	250.0	625.0	2 500.0	6 250.0
3	对羟基苯乙酮	0.12	0.30	1.20	3.00	12.00	30.00
4	苯氧乙醇	2.0	5.0	20.0	50.0	200.0	500.0
5	覆盆子酮	0.6	1.5	6.0	15.0	60.0	150.0
6	苯甲酸	0.30	0.75	3.00	7.50	30.00	75.00
7	4-羟基苯甲酸甲酯	0.2	0.5	2.0	5.0	20.0	50.0
8	对茴香酸	0.30	0.75	3.00	7.50	30.00	75.00
9	氯苯甘醚	0.2	0.5	2.0	5.0	20.0	50.0
10	脱氢乙酸	0.30	0.75	3.00	7.50	30.00	75.00
11	5-溴-5-硝基-1,3-二噁烷	2.50	6.25	25.00	62.50	250.00	625.00
12	辛酰羟肟酸	2.0	5.0	20.0	50.0	200.0	500.0
13	羟基乙氧基苯基丁酮	0.6	1.5	6.0	15.0	60.0	150.0

时间/min	φ（流动相A）/%	φ（流动相B）/%
0.0	95	5
4.0	95	5
4.1	80	20
25.0	70	30
25.1	65	35
27.0	65	35
27.1	95	5
30.0	95	5

序号	组分名称	ESI+/ESI-	母离子/（m/z）	子离子/（m/z）	去簇电压/V	碰撞电压/V
1	甲基异噻唑啉酮	+	116.0	71.1/101.1	70	28/30
2	乙酰丙酸	-	115.0	71.1/97.0	60	15/13
3	对羟基苯乙酮	-	135.1	92.0/93.0	80	31/25
4	覆盆子酮	-	163.1	57.0/163.1	40	24/8
5	苯甲酸	-	121.0	77.0/121.0	130	15/10
6	4-羟基苯甲酸甲酯	-	150.7	92.0/135.8	75	24/18
7	对茴香酸	-	151.1	107.1/92.0	60	16/35
8	脱氢乙酸	-	167.0	83.0/123.1	40	22/14
9	辛酰羟肟酸	-	158.1	58.0/110.2	90	17/17
10	羟基乙氧基苯基丁酮	-	206.8	57.0/179.1	60	31/14

序号	组分名称	线性方程	相关系数r	检出限/（mg/kg）	定量限/（mg/kg）
1	甲基异噻唑啉酮	y = 19.003 4 x+0.207 8	0.999 9	0.4	1.2
2	乙酰丙酸	y = 0.093 5 x+0.327 5	0.999 9	100.0	250.0
3	对羟基苯乙酮	y = 31.853 1 x+0.439 1	0.999 9	0.4	1.2
4	苯氧乙醇	y = 2.458 3 x+0.806 6	0.999 9	6.0	20.0
5	覆盆子酮	y = 7.587 5 x+0.788 7	0.999 9	2.0	6.0
6	苯甲酸	y = 27.268 7 x+2.104 0	0.999 9	1.0	3.0
7	4-羟基苯甲酸甲酯	y = 12.770 2 x+0.385 9	0.999 9	0.8	2.0
8	对茴香酸	y = 12.891 1 x+0.670 3	0.999 9	1.0	3.0
9	氯苯甘醚	y = 15.571 8 x+0.241 9	0.999 9	0.6	2.0
10	脱氢乙酸	y = 15.415 7 x-1.730 2	0.999 9	1.0	3.0
11	5-溴-5-硝基-1,3-二噁烷	y = 1.479 2 x+0.566 9	0.999 9	8.0	25.0
12	辛酰羟肟酸	y = 2.972 1 x-1.230 5	0.999 9	7.5	20.0
13	羟基乙氧基苯基丁酮	y = 3.442 5 x+0.277 2	0.999 9	2.0	6.0

组分名称	水剂		乳液		蜡基
组分名称	回收率/%	RSD/%	回收率/%	RSD/%	回收率/%	RSD/%
甲基异噻唑啉酮	88.6~101.7	1.3~1.7	86.5~102.8	1.4~1.7	95.8~110.5	0.9~3.7
乙酰丙酸	97.3~ 99.5	1.6~4.7	94.1~ 99.2	1.1~4.8	92.6~107.3	1.5~3.4
对羟基苯乙酮	87.6~101.1	1.2~1.9	88.2~105.7	1.0~3.2	97.4~111.5	1.8~2.7
苯氧乙醇	88.4~101.0	1.1~2.6	86.9~103.5	0.8~1.5	95.5~110.1	2.0~2.4
覆盆子酮	87.1~100.7	0.5~1.7	88.4~103.9	0.9~1.7	100.2~110.5	1.5~2.4
苯甲酸	86.0~102.5	0.8~1.7	91.2~104.7	1.0~3.4	96.9~107.2	2.0~2.7
4-羟基苯甲酸甲酯	87.7~100.4	1.5~3.0	90.1~103.7	0.9~4.1	96.6~115.4	1.8~2.8
对茴香酸	87.6~ 99.8	1.2~1.7	87.6~103.5	0.8~2.4	94.5~108.5	2.2~2.5
氯苯甘醚	88.3~100.6	1.2~1.6	88.6~103.7	0.9~2.4	93.5~110.3	1.8~2.6
脱氢乙酸	88.3~107.2	1.1~1.9	86.5~102.3	1.1~2.2	95.8~107.2	1.3~2.5
5-溴-5-硝基-1,3-二噁烷	87.4~100.0	1.2~3.0	84.1~102.2	1.0~1.3	84.0~109.5	1.7~2.7
辛酰羟肟酸	91.6~100.2	1.3~2.0	85.8~102.1	1.3~3.0	94.5~108.3	1.2~2.8
羟基乙氧基苯基丁酮	92.0~101.2	1.5~3.3	90.4~104.2	1.1~2.6	96.7~110.6	1.7~3.2