高效液相色谱法用于氧化型染发产品中42种限用染发剂的筛查和定量分析

doi:10.3969/j.issn.2097-2806.2024.04.016

摘要/Abstract

摘要：

建立了一种利用高效液相色谱筛查和定量分析氧化型染发产品中42种限用染发剂的方法。染发产品经亚硫酸氢钠水溶液-乙醇超声提取后，以0.02 mol/L乙酸铵水溶液-乙腈为流动相，在TechMate AQUA-STIII色谱柱上进行梯度洗脱和分离，选择235和280 nm作为检测波长，对42种染发剂进行一次性定性筛查，同时实现其中35种染发剂的定量分析，当筛查结果表明产品中含有2,6-二甲氧基-3,5-吡啶二胺盐酸盐、2-甲基-5-羟乙氨基苯酚等7种染发剂时，以0.02 mol/L乙酸铵水溶液-乙腈为流动相，在Waters Symmetry C₁₈色谱柱上进行等度洗脱，选择235 nm作为检测波长进行定量分析。结果表明：42种染发剂的方法检出限为10~101 µg/g，定量限为33~337 µg/g。在相应的线性范围内，42种染发剂色谱峰面积与质量浓度线性关系良好，相关系数（r）均大于0.999 8。在4个不同含量水平下，各目标化合物的平均回收率在81.3%~119.6%之间，RSD不大于5%。

关键词: 高效液相色谱法, 氧化型染发产品, 限用染发剂

Abstract:

A method for qualitative screening and quantitative analysis of 42 restricted hair dyes in oxidative hair color products by high performance liquid chromatography （HPLC） was established. The sample was ultrasonically extracted in mixed solvent of aqueous sodium bisulfite solution and ethanol. Firstly, the liquid chromatography condition for qualitative screening of 42 dyes and for quantitative analysis of 35 of them was established, which was as follows: The mobile phase was the mixed solvent of aqueous solution of 0.02 mol/L ammonium acetate and acetonitrile; gradient elution was carried out on a TechMate AQUA-STIII column; the detection wavelengths were 235 nm and 280 nm. If the other seven dyes including 2,6-dimethoxy-3,5-pyridinediamine·HCl, 2-methyl-5-hydroxyethylaminophenol, 3-nitro-p-hydroxyethylaminophenol, pyrazolone methyl pyrazolone, hydroxypropyl bis（N-hydroxyethyl-p-phenylenediamine）·HCl, 4-chlororesorcinol and 6-hydroxyindole were detected in the products, another liquid chromatography condition was required for their quantitative analysis, which was as follows: The mobile phase was the mixed solvent of aqueous solution of 0.02 mol/L ammonium acetate and acetonitrile; isocratic elution was carried out on a Waters Symmetry C₁₈column; the detection wavelength was 235 nm. The results showed that the limits of detection （LODs） of the 42 dyes were from 10 µg/g to 101 µg/g, while the limits of quantification （LOQs） were from 33 µg/g to 337 µg/g. The 42 dyes all had good linearities in their respective range, with correlation coefficients （r） higher than 0.999 8. The average spiked recoveries of 42 dyes at four levels were from 81.3% to 119.6%, with RSDs no more than 5%.

Key words: high performance liquid chromatography （HPLC）, oxidative hair color product, restricted hair dye

中图分类号:

TQ658

左雪, 邸铮, 汪亚菁, 杨玲, 张华珺, 柳青. 高效液相色谱法用于氧化型染发产品中42种限用染发剂的筛查和定量分析[J]. 日用化学工业（中英文）, 2024, 54(4): 490-498.

Xue Zuo, Zheng Di, Yajing Wang, Ling Yang, Huajun Zhang, Qing Liu. Qualitative screening and quantitative analysis of 42 restricted hair dyes in oxidative hair color products by high performance liquid chromatography[J]. China Surfactant Detergent & Cosmetics, 2024, 54(4): 490-498.

图/表 4

表1

42种染发剂标准物质信息"

序号	组分名称（组分编号）	标准物质来源	标准物质纯度/%
1	1-羟乙基-4,5-二氨基吡唑硫酸盐（C1）	Ark Pharm, Inc.（美国）	97
2	四氨基嘧啶硫酸盐（C2）	北京百灵威科技有限公司	98.9
3	2-氨基-3-羟基吡啶（C3）	北京曼哈格生物科技有限公司	99.9
4	对苯二胺（C4）	Dr. Ehrenstorfer GmbH（德国）	99.90
5	羟乙基对苯二胺硫酸盐（C5）	Ark Pharm, Inc.（美国）	95
6	对氨基苯酚（C6）	Dr. Ehrenstorfer GmbH（德国）	98.59
7	2,6-二氨基吡啶（C7）	Dr. Ehrenstorfer GmbH（德国）	99.4
8	甲苯-2,5-二胺硫酸盐（C8）	北京曼哈格生物科技有限公司	98.25
9	2,4-二氨基苯氧基乙醇盐酸盐（C9）	北京曼哈格生物科技有限公司	98.3
10	间氨基苯酚（C10）	北京曼哈格生物科技有限公司	99.64
11	4-氨基间甲酚（C11）	北京曼哈格生物科技有限公司	99.0
12	间苯二酚（C12）	Dr. Ehrenstorfer GmbH（德国）	99.83
13	N,N-双（2-羟乙基）-对苯二胺硫酸盐（C13）	北京曼哈格生物科技有限公司	99.9
14	对甲基氨基苯酚硫酸盐（C14）	北京曼哈格生物科技有限公司	99.5
15	2-甲基间苯二酚（C15）	北京曼哈格生物科技有限公司	99.7
16	2-氨基-6-氯-4-硝基苯酚（C16）	Sinco Pharmachem（美国）	97.47
17	羟苯并吗啉（C17）	Ark Pharm, Inc.（美国）	95
18	2-氨基-4-羟乙氨基茴香醚硫酸盐（C18）	Toronto research chemicals（加拿大）	99.12
19	4-氨基-2-羟基甲苯（C19）	北京曼哈格生物科技有限公司	99.1
20	6-甲氧基-2-甲氨基-3-氨基吡啶盐酸盐（C20）	Toronto research chemicals（加拿大）	96
21	4-硝基邻苯二胺（C21）	北京曼哈格生物科技有限公司	98.3
22	4-氨基-3-硝基苯酚（C22）	北京曼哈格生物科技有限公司	99.8
23	6-氨基间甲酚（C23）	北京曼哈格生物科技有限公司	99.7
24	2,6-二羟乙基氨甲苯（C24）	Sinco Pharmachem（美国）	95.58
25	2,6-二甲氧基-3,5-吡啶二胺盐酸盐（C25）	Toronto research chemicals（加拿大）	97
26	2-甲基-5-羟乙氨基苯酚（C26）	Toronto research chemicals（加拿大）	98.97
27	3-硝基对羟乙氨基酚（C27）	Sinco Pharmachem（美国）	99.47
28	苯基甲基吡唑啉酮（C28）	北京曼哈格生物科技有限公司	99.9
29	羟丙基双（N-羟乙基对苯二胺）盐酸盐（C29）	Toronto research chemicals（加拿大）	95
30	羟乙基-3,4-亚甲二氧基苯胺盐酸盐（C30）	Ark Pharm, Inc.（美国）	95
31	4-氯间苯二酚（C31）	北京曼哈格生物科技有限公司	99.0
32	6-羟基吲哚（C32）	北京曼哈格生物科技有限公司	99.2
33	5-氨基-6-氯-邻甲酚（C33）	Sinco Pharmachem（美国）	96.79
34	1,3-双-（2,4-二氨基苯氧基）丙烷盐酸盐（C34）	Sinco Pharmachem（美国）	96.68
35	4-羟丙氨基-3-硝基苯酚（C35）	Toronto research chemicals（加拿大）	99.70
36	1,5-萘二酚（C36）	北京曼哈格生物科技有限公司	99.5
37	2,7-萘二酚（C37）	北京曼哈格生物科技有限公司	99.3
38	HC黄2号（C38）	Sinco Pharmachem（美国）	99.9
39	5-氨基-4-氯邻甲酚（C39）	Sinco Pharmachem（美国）	96.9
40	羟乙基-2-硝基对甲苯胺（C40）	Sinco Pharmachem（美国）	97.6
41	N-苯基-对苯二胺（C41）	北京曼哈格生物科技有限公司	97.0
42	1-萘酚（C42）	Dr. Ehrenstorfer GmbH（德国）	99.6

表1

图1

图2

表2

42种染发剂回归方程、线性范围、检出限、定量限和加标回收率（n=6）"

序号^（1）	保留时间^（2）/min	线性方程	线性范围/（mg/L）	检出限/ （mg/g）	定量限/ （μg/g）	曲线最低点浓度水平		1 250 μg		2 500 μg		5 000 μg
序号^（1）	保留时间^（2）/min	线性方程	线性范围/（mg/L）	检出限/ （mg/g）	定量限/ （μg/g）	平均回收率/%	RSD/ %	平均回收率/%	RSD/ %	平均回收率/%	RSD/ %	平均回收率/%	RSD/ %
1	3.442	A=4.708 ρ-2.327	2.008~401.6	25	83	109.8	2.7	94.9	0.4	95.1	0.6	95.8	0.6
2	3.664	A=10.21 ρ-5.959	2.016~403.2	10	33	116.2	3.1	92.1	2.8	103.3	2.7	92.5	1.7
3	5.012	A=8.167 ρ+3.532	2.006~300.9	25	83	98.2	4.4	106.5	1.5	106.2	1.0	104.8	2.3
4	5.454	A=21.86 ρ-1.469	2.018~302.7	10	33	99.7	3.0	100.4	2.1	99.9	1.2	99.2	2.5
5	6.201	A=9.095 ρ+0.4296	1.969~295.3	25	83	91.1	4.4	101.3	1.9	100.3	1.3	98.6	3.0
6	7.018	A=18.22 ρ+3.414	2.004~400.8	10	33	91.6	1.0	101.1	1.8	103.5	1.4	95.4	4.1
7	8.301	A=18.13 ρ-8.501	5.000~400.0	10	33	108.4	1.4	104.3	0.7	100.0	1.3	101.5	2.0
8	9.771	A=9.844 ρ-1.464	2.016~403.2	25	83	104.1	1.4	90.5	2.9	96.5	2.6	95.5	2.6
9	10.973	A=9.914 ρ+0.9675	2.012~402.4	25	83	96.5	3.5	101.5	0.1	102.8	0.4	101.4	0.5
10	11.608	A=17.70 ρ-6.344	2.018~403.6	10	33	118.1	0.4	103.3	1.0	99.6	1.2	101.0	2.0
11	13.278	A=14.98 ρ-4.017	1.988~397.5	25	83	102.9	2.1	97.2	2.3	96.1	0.9	91.3	4.2
12	16.886	A=4.261 ρ-2.054	5.005~400.4	25	83	110.7	2.2	102.8	1.3	99.0	1.2	100.4	2.1
13	19.427	A=4.929 ρ-3.726	5.060~404.8	101	337	92.7	2.8	104.6	3.8	109.0	1.3	107.8	3.0
14	20.363	A=13.47 ρ-7.523	1.996~399.1	25	83	113.7	2.9	90.1	3.2	100.1	1.6	84.9	1.6
15	21.776	A=3.785 ρ-1.454	5.045~403.6	101	337	107.0	2.5	99.0	1.9	97.7	1.4	99.8	2.3
16	27.298	A=15.01 ρ-6.787	1.988~198.8	25	83	98.3	2.0	105.9	2.6	107.3	0.3	109.7	2.2
17	30.650	A=9.351 ρ-2.929	1.954~390.8	24	80	116.5	1.9	96.2	4.0	99.1	1.5	97.5	3.8
18	31.847	A=7.305 ρ-3.776	2.032~406.4	25	83	119.6	0.6	109.7	1.5	113.1	1.4	112.7	1.6
19	32.752	A=16.10 ρ-8.250	5.015~401.2	25	83	110.3	2.5	103.7	4.4	100.3	0.7	100.6	2.7
20	33.975	A=6.425 ρ-0.6760	4.810~384.8	48	160	87.1	3.1	90.8	2.3	98.7	1.4	91.7	3.0
21	34.871	A=13.93 ρ-0.3016	2.006~401.2	25	83	95.8	2.2	97.9	2.4	96.5	2.0	97.6	2.3
22	35.596	A=29.85 ρ-7.754	2.020~404.0	25	83	107.6	2.1	95.5	2.6	96.3	1.7	98.4	2.4
23	36.609	A=14.61 ρ-4.333	2.002~400.4	25	83	109.0	1.5	92.7	1.9	98.7	1.6	94.7	4.5
24	40.004	A=27.46 ρ-3.112	1.927~385.4	10	33	107.0	1.2	105.0	0.7	101.1	1.1	102.4	2.0
25	42.853 （15.334）	A=7.895 ρ+8.340	4.941~395.3	25	83	81.3	1.1	87.2	4.3	89.3	2.7	85.6	4.9
26	43.267 （14.475）	A=13.46 ρ+2.952	2.012~402.4	10	33	92.4	4.1	92.9	4.9	99.7	0.5	108.6	1.9
27	43.542 （11.484）	A=28.13 ρ+6.119	2.026~405.2	10	33	90.0	2.9	93.3	3.7	99.8	0.9	107.2	1.4
28	44.379 （17.221）	A=17.29 ρ+3.545	2.026~405.2	25	83	82.3	2.2	83.5	3.2	89.1	2.2	89.0	3.3
29	45.914 （8.787）	A=4.934 ρ-7.659	5.060 ~404.8	51	170	88.6	2.3	93.9	4.9	84.5	2.0	99.3	5.0
30	46.226	A=6.131 ρ-2.231	4.807~384.6	48	160	111.2	3.2	105.5	1.1	99.5	1.0	101.4	2.0
31	48.062 （22.545）	A=7.341 ρ+1.299	2.010~402.0	25	83	96.7	4.3	93.2	1.8	100.1	0.6	106.2	2.4
32	48.498 （23.515）	A=8.158 ρ+1.267	2.006~401.2	25	83	98.3	4.7	95.1	2.1	98.1	0.7	105.0	1.8
33	50.742	A=14.38 ρ-7.266	4.902~392.2	25	83	106.4	2.7	97.0	2.1	97.2	1.7	95.1	0.8
34	53.257	A=9.231 ρ-3.854	1.954~390.8	49	163	111.8	1.2	104.1	2.2	105.6	1.2	105.2	2.8
35	54.054	A=27.17 ρ+3.961	2.018~403.6	25	83	92.1	2.5	94.3	1.5	95.3	1.8	98.8	2.1
36	55.820	A=28.98 ρ+0.2450	2.008~301.2	10	33	95.9	2.6	84.7	3.8	90.1	2.8	86.7	3.6
37	59.815	A=5.796 ρ-0.7417	2.014~402.8	25	83	100.7	3.1	90.6	3.5	93.3	1.8	105.9	1.3
38	61.180	A=7.039 ρ+2.978	2.056~411.2	10	33	84.2	3.3	101.9	2.0	99.1	1.5	103.8	1.7
39	61.613	A=2.366 ρ+0.5020	4.900~392.0	25	83	96.0	2.9	97.2	2.3	100.5	1.7	103.1	3.5
40	62.649	A=6.781 ρ-0.1798	1.993~299.0	10	33	103.1	2.9	101.3	2.5	96.6	1.2	98.9	2.5
41	63.062	A=24.97 ρ-4.950	1.993~299.0	10	33	109.9	3.8	97.4	3.2	93.2	2.9	95.7	2.2
42	63.549	A=7.120 ρ-2.532	2.010~402.0	25	83	105.4	4.2	86.5	1.4	88.5	3.3	89.3	3.8

表2

参考文献 22

[1]	Wu Jing, Liu Min, Xing Shuxia, et al. Quality and safety analysis of hair dyeing cosmetics during 2018 to 2019[J]. Flavour Fragrance Cosmetics, 2021 (1) : 65-68, 72.
[2]	Wang Hui, Tian Siyu, Yin Yi, et al. Quality analysis and supervision suggestions of cosmetic label[J]. Food and Drug, 2021, 23 (1) : 64-68.
[3]	Lin Lin, Gu Yuxiang, Wang Dinglin. Current status of hair dye detection methods and analysis on unqualified products in national inspection[J]. Flavour Fragrance Cosmetics, 2020 (3) : 66-69, 81.
[4]	Xia Aiting, Li Chenxi, Zhang Jiaojiao, et al. The research on sensitizing ingredients in oxidative hair dye products on the Chinese market and common allergens in patients with hair dye allergy[J]. The Chinese Journal of Dermatovenereology, 2019, 33 (8) : 938-942.
[5]	Hanseul P, Jee-hyun H, Ji-Seok H, et al. Skin irritation and sensitization potential of oxidative hair dye substances evaluated with in vitro, in chemico and in silico test methods[J]. Food and Chemical Toxicology, 2018, 121: 360-366. doi: S0278-6915(18)30661-6 pmid: 30213554
[6]	Zanoni T B, Pedrosa T N, Catarino C M, et al. Allergens of permanent hair dyes induces epidermal damage, skin barrier loss and IL-1α increase in epidermal in vitro model[J]. Food and Chemical Toxicology, 2018, 112: 265-272. doi: 10.1016/j.fct.2017.12.033
[7]	State Food and Drug Administration. Safety and technical standard for cosmetics (2015 Edition) [S]. Beijing: Standard Press of China, 2016.
[8]	GB/T 35824—2018. Determination of 20 kinds of prohibited and restricted dyestuff in hair dyes-High performance liquid chromatography [S]. Beijing: Standard Press of China, 2018.
[9]	Liu Yun, Liu Jie, Huang Wei, et al. Determination of 15 kinds of dyestuffs in hair dyes by high performance liquid chromatography[J]. China Surfactant Detergent & Cosmetics, 2022, 52 (12) : 1333-1341.
[10]	Jiang Chengjun, Wu Zhishan, Huang Haiping, et al. Simultaneous determination of 10 kinds of dyestuffin prohibited in hair dye cosmetics by high performance liquid chromatography[J]. Flavour Fragrance Cosmetics, 2022 (5) : 44-48.
[11]	Li Hongying, Huang Cheng, Ding Xiaoping, et al. Determination of 21 kinds of banned and restricted dyestuffs in hair dyes by high performance liquid chromatography[J]. Flavour Fragrance Cosmetics, 2019 (4) : 28-32, 46.
[12]	Zhang Guocui, Lu Yichuan, Li Chunling. Simultaneous determination of 26 dyes in hair colorants by high performance liquid chromatography[J]. Flavour Fragrance Cosmetics, 2019 (5) : 43-47.
[13]	Wu Silin, Peng Xingsheng, Zheng Rong. Simultaneous determination of 12 hair dyes in cosmetics by performance chromatography and verification by liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Health Laboratory Technology, 2022, 32 (14) : 1665-1670.
[14]	Lu Duanping, Chen Xu, Tang Wen, et al. Determination of 44 kinds of dyestuffs in oxidative hair dye products by HPLC[J]. Journal of Pharmaceutical Analysis, 2021, 41 (12) : 2191-2201.
[15]	Yang Danmei, Shi Xinghogn, Luo Jinmei, et al. Simultaneous determination of 5 restricted dyes in hair dyeing cosmetics by high performance liquid chromatography[J]. China Surfactant Detergent & Cosmetics, 2022, 52 (1) : 103-108.
[16]	Zuo Xue, Di Zheng, Du Yong, et al. Determination of 40 dyes in oxidative hair dye products by high performance liquid chromatography[J]. Chinese Journal of Chromatography, 2021, 39 (11) : 1222-1229. doi: 10.3724/SP.J.1123.2020.11020
[17]	Li Ruoqi, Wu Rui, Sun Ying, et al. Determination of 32 kinds of component in hair dyes by HPLC-MS/MS and stability analysis of dye intermediates[J]. China Surfactant Detergent & Cosmetics, 2021, 51 (12) : 1269-1277.
[18]	Zheng Yi, Chen Lisong, Cheng Shan, et al. Simultaneous determination of ten hair dyes in hair-dyeing cosmetics by ultra performance liquid chromatography-tandem mass spectrometry[J]. Flavour Fragrance Cosmetics, 2021 (3) : 90-96.
[19]	Ju Lingyan, Li Zhaojie, Song Weide, et al. Rapid screening of 32 dyes in oxidative hair dyes by ultra-performance liquid chromatography-quadrupole/electrostatic field obitrap high-resolution mass spectrometry[J]. Journal of Instrumental Analysis, 2019, 38 (8) : 931-938.
[20]	Zhong Zhixiong, Luo Zhibin, Zhu Binghui. Development of ultrasound-assisted matrix solid-phase dispersive liquid extraction method for determination of nine intermediates in hair dyes by ion chromatography[J]. Chinese Journal of Inorganic Analytical Chemistry, 2012, 2 (3) : 31-34.
[21]	Zhu Ran, Zhang Xulong, Wang Jide, et al. GC determination of 13dyes and impurities in hair colorant[J]. Physical Testing and Chemical Analysis (Part B: Chemical Analysis), 2012, 48 (5) : 526-529.
[22]	Guerra E, Pablo L J, Llompart M, et al. Determination of oxidative hair dyes using miniaturized extraction techniques and gas chromatography-tandem mass spectrometry[J]. Microchemical Journal, 2017, 132: 308-318. doi: 10.1016/j.microc.2017.02.017