高效液相色谱法测定氧化型染发产品中47种染发剂

doi:10.3969/j.issn.2097-2806.2023.06.016

摘要/Abstract

摘要：

建立了测定氧化型染发产品中47种染发剂含量的高效液相色谱分析方法。样品经无水乙醇-10 g/L亚硫酸氢钠水溶液（体积比7∶3）超声提取后，加入2 g/L亚硫酸氢钠水溶液定容至25 mL。采用Shim-pack GIST C₁₈ （4.6 mm×250 mm，5 μm）色谱柱分离，分别以乙腈-20 mmol/L磷酸二氢钾溶液（pH7.5）、甲醇-20 mmol/L磷酸二氢钾溶液（pH7.5）为流动相梯度洗脱，流速1.0 mL/min，柱温25 ℃，检测波长230和280 nm，标准曲线法定量。结果表明，47种染发剂在相应的浓度范围线性关系良好，相关系数均大于0.999，检出限（LOD）为1~35 μg/g。在3种加标水平下的平均加标回收率为80.4%~110.7%，相对标准偏差为0.25%~5.08%。与现行标准检验方法相比，该方法增加了15种准用染发剂的测定，具有较高的准确度和精密度，能满足实际检测需求，适用于氧化型染发产品中47种染发剂的测定。

关键词: 高效液相色谱法, 氧化型染发产品, 染发剂, 含量测定

Abstract:

A high performance liquid chromatographic （HPLC） method was developed for the determination of 47 kinds of dyestuffs in oxidative hair dye. The samples were extracted ultrasonically with anhydrous ethanol-10 g/L sodium bisulfite solution （7∶3）, and then 2 g/L sodium bisulfite solution was added to a constant volume of 25 mL. If necessary, the extractions were centrifuged at 10 000 r/min for 5 minutes. Chromatographic separation was carried on a Shim-pack GIST C₁₈ （4.6 mm×250 mm, 5 μm） column by gradient elution. Acetonitrile-20 mmol/L dipotassium hydrogen phosphate （pH7.5） and methanol-20 mmol/L dipotassium hydrogen phosphate （pH7.5） were used as the mobile phase, respectively. The flow rate was 1.0 mL/min and the column temperature was 25 ℃. The analysis of 47 kinds of dyestuffs was performed by HPLC with diode array detector at the wavelength of 230, 280 nm, respectively. Quantification analysis was performed by the calibration curve method. The linear ranges of 47 kinds of dyestuffs show good linearity with the correlation coefficients more than 0.999. The limits of detection （LOD） are in the ranges of 1-35 μg/g. The mean recoveries at three different concentration levels are 80.4%-110.7% with the RSDs of 0.25%-5.08%. Compared with the current standard, this method increases the determination of 15 permitted dyes. This method has high accuracy and precision, and can meet the actual detection needs. It is suitable for the determination of 47 kinds of dyestuffs in oxidized hair dyes.

Key words: high performance liquid chromatography, oxidized hair dyes, dyestuff, content determination

中图分类号:

TQ658

高瑞芳, 柴格, 李祥胜. 高效液相色谱法测定氧化型染发产品中47种染发剂[J]. 日用化学工业（中英文）, 2023, 53(6): 725-732.

Gao Ruifang, Chai Ge, Li Xiangsheng. Determination of 47 kinds of dyestuffs in oxidative hair dye by high performance liquid chromatography[J]. China Surfactant Detergent & Cosmetics, 2023, 53(6): 725-732.

图/表 6

表1

47种染发剂标准品来源"

分组编号	标准品名称	来源	分组编号	标准品名称	来源
A1	1-羟乙基-4,5-二氨基吡唑硫酸盐	PANPHY	B1	四氨基嘧啶硫酸盐	Dr.Ehrenstorfer GmbH
A2	2-氨基-3-羟基吡啶	TRC	B2	间苯二胺	Dr.Ehrenstorfer GmbH
A3	对苯二胺	Dr.Ehrenstorfer GmbH	B3	氢醌	Dr.Ehrenstorfer GmbH
A4	羟乙基对苯二胺硫酸盐	PANPHY	B4	间氨基苯酚	Dr.Ehrenstorfer GmbH
A5	对氨基苯酚	Dr.Ehrenstorfer GmbH	B5	2,4-二氨基苯氧基乙醇盐酸盐	Be Pure
A6	2,6-二氨基吡啶	Dr.Ehrenstorfer GmbH	B6	邻苯二胺	Dr.Ehrenstorfer GmbH
A7	甲苯-2,5-二胺硫酸盐	Dr.Ehrenstorfer GmbH	B7	邻氨基苯酚	Dr.Ehrenstorfer GmbH
A8	4-氨基间甲酚	Dr.Ehrenstorfer GmbH	B8	对甲基氨基苯酚硫酸盐	PANPHY
A9	间苯二酚	Dr.Ehrenstorfer GmbH	B9	2-氨基-6-氯-4-硝基苯酚	First Standard
A10	2-氯对苯二胺硫酸盐	PANPHY	B10	N,N-双（2-羟乙基）对苯二胺硫酸盐	PANPHY
A11	2-甲基间苯二酚	SIGMA-ALDRICH	B11	4-硝基邻苯二胺	TRC
A12	2-硝基对苯二胺	Dr.Ehrenstorfer GmbH	B12	甲苯-3,4-二胺	Dr.Ehrenstorfer GmbH
A13	苯基甲基吡唑啉酮	PANPHY	B13	6-氨基间甲酚	First Standard
A14	羟苯并吗啉	PANPHY	B14	2,6-二羟乙基氨甲苯	First Standard
A15	4-氨基-2-羟基甲苯	TRC	B15	3-硝基对羟乙氨基酚	First Standard
A16	4-氨基-3-硝基苯酚	TRC	B16	2-甲基-5-羟乙氨基苯酚	PANPHY
A17	4-氯间苯二酚	TRC	B17	5-氨基-6-氯-邻甲酚	First Standard
A18	6-羟基吲哚	SIGMA-ALDRICH	B18	4-羟丙氨基-3-硝基苯酚	First Standard
A19	碱性红51号	PANPHY	B19	羟乙基-3,4-亚甲二氧基苯胺盐酸盐	Be Pure
A20	1,5-萘二酚	PANPHY	B20	2,7-萘二酚	TRC
A21	5-氨基-4-氯邻甲酚	Be Pure	B21	N,N-二乙基对苯二胺硫酸盐	Dr.Ehrenstorfer GmbH
A22	羟乙基-2-硝基对甲苯胺	First Standard	B22	HC黄2号	Be Pure
A23	1-萘酚	Dr.Ehrenstorfer GmbH	B23	N,N-二乙基甲苯-2,5-二胺盐酸盐	Dr.Ehrenstorfer GmbH
			B24	N-苯基对苯二胺	PANPHY

表1

表2

图1

图2

表3

47种染发剂的保留时间、线性方程、相关系数和检出限"

化合物	保留时间/min	线性方程	相关系数	检出限/（μg/g）	化合物	保留时间/min	线性方程	相关系数	检出限/（μg/g）
A1	3.290	A=45.80ρ-0.058 4	0.999 9	22	B1	3.647	A=128.2ρ-0.100 9	0.999 9	3
A2	4.807	A=102.7ρ+0.055 3	0.999 9	6	B2	9.080	A=51.04ρ+0.018 6	0.999 9	5
A3	6.083	A=213.7ρ-0.431 1	0.999 9	4	B3	10.433	A=67.19ρ-0.030 7	0.999 9	4
A4	6.967	A=89.77ρ-0.363 7	0.999 7	13	B4	12.280	A=64.43ρ+0.034 9	0.999 9	5
A5	7.720	A=223.9ρ-0.493 5	0.999 9	4	B5	13.347	A=19.53ρ+0.042 5	0.999 9	13
A6	9.767	A=186.1ρ-0.269 2	0.999 9	6	B6	16.020	A=74.24ρ+0.026 6	0.999 9	5
A7	11.690	A=102.4ρ-0.409 9	0.999 9	20	B7	18.947	A=86.87ρ+0.017 3	0.999 9	5
A8	15.623	A=181.0ρ-0.110 6	0.999 9	11	B8	23.803	A=20.01ρ+0.027 9	0.999 9	30
A9	20.740	A=91.29ρ+0.037 2	0.999 9	23	B9	24.730	A=152.5ρ+0.047 1	0.999 9	3
A10	22.610	A=87.49ρ-0.251 8	0.999 8	25	B10	26.223	A=30.92ρ-0.014 2	0.999 9	35
A11	24.203	A=113.0ρ+0.000 7	0.999 9	11	B11	32.267	A=115.4ρ+0.420 7	0.999 9	3
A12	25.447	A=322.0ρ-0.205 6	0.999 9	6	B12	35.823	A=52.01ρ-0.012 1	0.999 9	6
A13	26.637	A=150.9ρ-0.469 0	0.999 9	14	B13	38.187	A=66.12ρ+0.008 0	0.999 9	4
A14	27.310	A=163.0ρ+0.163 8	0.999 9	4	B14	40.100	A=16.08ρ+0.020 4	0.999 9	9
A15	29.223	A=207.3ρ+0.016 3	0.999 9	4	B15	40.530	A=96.93ρ+0.006 3	0.999 9	2
A16	30.617	A=391.7ρ-0.251 7	0.999 9	2	B16	40.997	A=41.98ρ+0.030 0	0.999 9	5
A17	34.027	A=144.1ρ+0.007 7	0.999 9	5	B17	41.850	A=43.44ρ+0.030 2	0.999 9	5
A18	35.410	A=259.8ρ+1.394 0	0.999 5	2	B18	43.470	A=89.11ρ+0.035 1	0.999 9	2
A19	36.067	A=29.01ρ-0.068 3	0.999 8	12	B19	44.700	A=37.88ρ-0.005 8	0.999 9	6
A20	36.503	A=404.6ρ+0.489 6	0.999 8	1	B20	45.097	A=82.31ρ+0.045 4	0.999 9	2
A21	38.080	A=184.4ρ+0.070 8	0.999 9	1	B21	46.617	A=26.30ρ+0.010 8	0.999 9	7
A22	39.280	A=342.2ρ+0.201 7	0.999 9	1	B22	47.560	A=95.85ρ+0.052 1	0.999 9	2
A23	42.403	A=756.7ρ+0.067 3	0.999 9	1	B23	51.250	A=32.67ρ-0.065 5	0.999 9	28
					B24	52.200	A=300.5ρ+0.075 5	0.999 9	1

表3

表4

样品的加标回收率及相对标准偏差（n=3）"

化合物	平均加标回收率/%			RSD/%			化合物	平均加标回收率/%			RSD/%
化合物	低	中	高	低	中	高	化合物	低	中	高	低	中	高
A1	103.0	97.0	92.4	0.85	2.48	2.19	B1	89.8	96.2	88.9	3.65	3.92	2.07
A2	90.4	92.6	92.4	1.72	2.21	3.79	B2	99.2	100.2	100.2	1.54	0.62	0.49
A3	105.5	103.6	103.1	0.47	1.42	1.35	B3	92.3	93.9	83.1	3.57	4.39	2.51
A4	105.4	101.5	107.7	1.35	0.60	1.75	B4	98.9	100.5	101.4	0.96	0.83	0.79
A5	99.8	99.5	99.6	2.66	2.66	3.38	B5	91.0	90.3	89.3	0.47	1.74	1.49
A6	105.8	106.7	104.8	2.52	1.22	1.59	B6	97.4	99.7	100.7	1.37	0.36	0.89
A7	107.2	101.2	98.6	1.61	1.79	1.37	B7	98.6	99.9	100.8	1.31	0.92	0.82
A8	93.1	100.3	96.0	4.42	4.87	1.69	B8	90.8	100.9	91.8	4.81	4.95	5.00
A9	102.7	104.2	104.9	1.25	0.99	0.54	B9	97.1	98.7	101.0	0.47	2.29	0.78
A10	110.7	104.4	101.3	0.55	1.43	3.10	B10	93.1	97.4	97.9	2.76	0.98	0.46
A11	107.0	106.9	106.5	0.95	1.62	1.90	B11	87.5	94.4	99.3	1.87	1.61	0.36
A12	106.1	105.0	104.3	0.38	1.00	0.70	B12	95.6	99.7	99.8	2.79	0.37	0.91
A13	99.0	94.9	91.8	2.16	3.65	2.39	B13	98.8	99.4	99.8	2.16	0.52	0.64
A14	102.5	102.0	100.5	1.31	0.74	0.86	B14	97.8	99.5	100.8	1.29	1.61	0.67
A15	103.3	102.9	101.1	1.08	1.13	0.70	B15	99.7	100.4	101.6	2.08	1.58	1.04
A16	100.5	103.0	103.4	2.63	0.95	0.43	B16	101.3	102.2	103.5	1.08	1.80	1.01
A17	99.9	101.8	101.5	1.90	0.88	0.33	B17	96.7	96.0	97.4	1.29	3.34	1.63
A18	89.3	104.1	107.8	3.38	0.94	0.51	B18	97.6	99.4	100.6	1.77	1.33	0.87
A19	106.1	107.9	100.0	2.07	4.50	4.33	B19	100.3	101.0	100.4	2.98	0.25	0.79
A20	80.4	83.3	82.1	1.72	2.92	4.72	B20	86.9	89.7	93.0	2.14	5.08	4.54
A21	100.2	102.5	102.5	2.46	0.86	0.51	B21	96.8	98.3	101.1	1.51	1.25	0.87
A22	101.9	104.5	104.4	1.54	0.91	0.59	B22	98.7	99.8	102.6	1.98	1.20	1.18
A23	88.4	97.6	96.4	3.99	1.36	1.08	B23	102.6	97.6	96.5	2.60	2.57	0.32
							B24	97.9	98.5	101.0	2.08	2.30	1.16

表4

参考文献 14

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方法	1,5-萘二酚含量/%
方法	0 h	3 h	8 h	14 h	20 h
自建方法	100	99.0	98.1	97.5	96.6
《规范》方法	100	79.4	34.4	6.6	0.4

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