UPLC-MS/MS法测定化妆品常用植物提取物中16种香豆素类化合物

doi:10.3969/j.issn.1001-1803.2023.02.014

摘要/Abstract

摘要：

建立了同时测定常用化妆品植物提取物原料中16种香豆素类化合物的UPLC-MS/MS法，为评估植物提取物作为化妆品原料的安全性和规范植物提取物在化妆品中的应用提供理论依据。采用液质联用法，应用Waters ACQUITY HPLC BEH C₁₈柱，以乙腈-0.1%甲酸溶液为流动相，梯度洗脱分离，质谱采用正离子模式，多反应监测，流速为0.3 mL/min。16种香豆素类化合物在各自质量浓度范围内呈现良好的线性关系，相关系数均大于0.995。在低、中、高3个加标水平的平均回收率为85.7%~108.6%，相对标准偏差为1.5%~5.8%，16种香豆素类化合物检出限为0.3~28.0 μg/kg，定量限为1.0~93.0 μg/kg，收集的14批植物提取物中，4批检出香豆素类化合物。该方法准确、分析速度快、操作简便、灵敏度高，能满足化妆品植物提取物原料中16种香豆素类化合物的定性筛查和定量检测要求，为探究植物提取物致敏风险物质基础提供一种思路，建议关注化妆品植物提取物原料带入致敏成分的安全风险情况，建立化妆品植物提取物原料质量控制和使用规范。

关键词: 超高效液相色谱-串联质谱法, 化妆品, 植物提取物, 香豆素类化合物

Abstract:

An analytical method based on UPLC-MS/MS for the determination of 16 coumarin compounds in commonly cosmetics plant extract raw materials was established. The study aimed to evaluate the safety of plant extract as raw materials for cosmetics and to provide theoretical basis for regulating the application of plant extract in cosmetics. The separation was performed on a Waters ACQUITY HPLC BEH C₁₈ column. Acetonitrile and 0.1% formic acid were used as mobile phases with gradient elution at a flow rate of 0.30 mL/min. A triple quadrupole mass spectrometer operating in positive electrospray ionization mode with multiple reaction monitoring was used. The results of 16 coumarin compounds show good linear relationships within their own rangers, the correlation coefficients （r） are above 0.995, and the mean recoveries at three spiked levels range from 85.7% to 108.6, with the relative standards of 1.5%-5.8%. The limits of detections （LODS） for the 16 coumarin compounds range from 0.3 μg/kg to 28.0 μg/kg. The limits of quantitation （LOQS） range from 1.0 μg/kg to 93.0 μg/kg. Coumarin compounds are detected in 4 of the 14 batches of plant extracts collected. The method not only is convenient, rapid, sensitive and accurate, but also can be used for the qualitative screening and quantitative determination of 16 coumarin compounds in cosmetics plant extract raw materials, which provides a way to explore the material basis of allergenic risk of the plant extract raw materials. It is suggested to pay attention to the safety risk of allergenic components from the plant extract raw materials, to establish the product quality and use specification of the plant extract raw materials.

Key words: UPLC-MS/MS, cosmetic, plant extract, coumarin compound

中图分类号:

TQ658

张艳萍, 周宗洲, 黄婉锋, 陈艺华, 张敏, 何作民. UPLC-MS/MS法测定化妆品常用植物提取物中16种香豆素类化合物[J]. 日用化学工业（中英文）, 2023, 53(2): 226-232.

Zhang Yanping, Zhou Zongzhou, Huang Wanfeng, Chen Yihua, Zhang Min, He Zuomin. Determination of 16 coumarin compounds in commonly cosmetics plant extract raw materials by UPLC-MS/MS[J]. China Surfactant Detergent & Cosmetics, 2023, 53(2): 226-232.

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参考文献 12

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序号	名称	来源	批号	纯度/%
1	香豆素	Bepure	D0009958	99.9
2	7-羟基香豆素	Chemservice	7642400	99.5
3	补骨脂素	Bepure	A0014475	99.6
4	7-甲基香豆素	Bepure	E0019273	99.5
5	异补骨脂素	Bepure	C0013514	99.8
6	8-甲氧基补骨脂素	Bepure	D0016009	99.9
7	欧前胡素	Bepure	B0014639	99.0
8	异欧前胡素	Bepure	B0009737	99.2
9	6，7-二甲氧基香豆素	Bepure	C0007144	99.7
10	新补骨脂异黄酮	中检院	520052-201401	99.6
11	补骨脂二氢黄酮	中检院	520053-201401	99.4
12	4-甲基-7-羟基香豆素	CATO	1018-RC-0004	100.0
13	4-甲基-7-乙氧基香豆素	Bepure	E0026326	99.9
14	5-甲氧基补骨脂素	Bepure	A0014497	99.6
15	2，4，8-三甲基补骨脂素	Bepure	C0013508	99.0
16	环香豆素	Dr.E	1063029	99.0

序号	名称	移取体积/mL	质量浓度/（μg/mL）
1	香豆素	4.00	80.0
2	7-羟基香豆素、补骨脂素、7-甲基香豆素、异补骨脂素、8-甲氧基补骨脂素	2.00	40.0
3	欧前胡素、异欧前胡素、6，7-二甲氧基香豆素、新补骨脂异黄酮、补骨脂二氢黄酮	0.40	8.0
4	4-甲基-7-羟基香豆素、4-甲基-7-乙氧基香豆素、5-甲氧基补骨脂素、2，4，8-三甲基补骨脂素、环香豆素	0.20	4.0

序号	名称	保留时间/min	母离子（m/z）	子离子（m/z）	锥孔电压/V	碰撞电压/V
1	香豆素	5.43	146.9	90.9*	32	20
1	香豆素	5.43	146.9	76.9	32	22
2	7-甲基香豆素	8.35	160.9	104.9*	34	20
2	7-甲基香豆素	8.35	160.9	90.8	34	20
3	7-羟基香豆素	4.00	162.9	106.9 *	34	22
3	7-羟基香豆素	4.00	162.9	76.8	34	28
4	4-甲基-7-羟基香豆素	4.76	176.9	120.9*	38	22
4	4-甲基-7-羟基香豆素	4.76	176.9	104.9	38	20
5	补骨脂素	8.21	186.9	130.9*	34	26
5	补骨脂素	8.21	186.9	76.8	34	36
6	异补骨脂素	8.47	186.9	130.8*	34	22
6	异补骨脂素	8.47	186.9	76.9	34	34
7	4-甲基-7-乙氧基香豆素	9.20	205.0	176.9*	32	20
7	4-甲基-7-乙氧基香豆素	9.20	205.0	104.9	32	22
8	6，7-二甲氧基香豆素	5.06	207.0	106.9*	36	34
8	6，7-二甲氧基香豆素	5.06	207.0	150.9	36	22
9	8-甲氧基补骨脂素	8.54	217.0	201.9*	36	22
9	8-甲氧基补骨脂素	8.54	217.0	89.5	36	36
10	5-甲氧基补骨脂素	8.92	217.0	202.0*	34	24
10	5-甲氧基补骨脂素	8.92	217.0	117.9	34	32
11	2，4，8-三甲基补骨脂素	9.73	229.1	141.8*	42	24
11	2，4，8-三甲基补骨脂素	9.73	229.1	173.0	42	22
12	异欧前胡素	10.14	271.1	203.0*	16	12
12	异欧前胡素	10.14	271.1	90.8	16	40
13	欧前胡素	9.79	271.1	146.9*	18	34
13	欧前胡素	9.79	271.1	202.9	18	12
14	环香豆素	10.44	323.2	120.9*	24	44
14	环香豆素	10.44	323.2	251.0	24	18
15	新补骨脂异黄酮	9.35	323.2	267.0*	32	18
15	新补骨脂异黄酮	9.35	323.2	255.0	32	20
16	补骨脂二氢黄酮	9.47	325.2	64.8*	34	44
16	补骨脂二氢黄酮	9.47	325.2	148.9	34	24

序号	名称	线性范围/（μg/mL）	回归方程	相关系数r	LOD/（μg/kg）	LOQ/（μg/kg）
1	香豆素	0.20~4.0	Y=16 241.9X+104.8	0.998 8	18.0	60.0
2	7-羟基香豆素	0.10~2.0	Y=45 779.9X-236.9	0.997 8	13.0	43.0
3	补骨脂素	0.10~2.0	Y=99 642.9X+561.4	0.999 4	28.0	93.0
4	7-甲基香豆素	0.10~2.0	Y=77 821.3X+452.9	0.998 1	23.0	77.0
5	异补骨脂素	0.10~2.0	Y=100 678X+231.7	0.999 4	27.0	90.0
6	8-甲氧基补骨脂素	0.10~2.0	Y=145 635X+298.1	0.999 5	10.0	33.0
7	欧前胡素	0.02~0.40	Y=172 310X+431.8	0.999 2	3.0	10.0
8	异欧前胡素	0.02~0.40	Y=81 886.4X+326.1	0.996 1	1.0	3.3
9	6，7-二甲氧基香豆素	0.02~0.40	Y=108 121X-542.3	0.998 0	1.0	3.3
10	新补骨脂异黄酮	0.02~0.40	Y=205 241X+412.6	0.999 8	0.7	2.2
11	补骨脂二氢黄酮	0.02~0.40	Y=136 484X-326.1	0.997 5	1.3	4.5
12	4-甲基-7-羟基香豆素	0.01~0.20	Y=47 302.3X+103.7	0.999 5	0.6	2.0
13	4-甲基-7-乙氧基香豆素	0.01~0.20	Y=527 688X+523.9	0.998 9	0.7	2.2
14	5-甲氧基补骨脂素	0.01~0.20	Y=388 853X-239.4	0.998 7	0.8	2.8
15	2，4，8-三甲基补骨脂素	0.01~0.20	Y=493 485X+329.5	0.998 4	0.3	1.0
16	环香豆素	0.01~0.20	Y=738 339X-204.2	0.998 6	0.6	2.0

化合物	理论含量/μg			平均回收率/%			RSD/%
化合物	低	中	高	低	中	高	低	中	高
香豆素	4.25	8.49	16.98	85.7	90.3	91.4	4.6	3.9	5.1
7-羟基香豆素	2.12	4.23	8.46	91.6	95.7	92.7	2.8	3.8	3.1
补骨脂素	2.22	4.44	8.88	90.4	92.1	94.7	5.8	4.3	1.9
7-甲基香豆素	2.20	4.40	8.80	88.4	89.4	101.5	2.8	3.7	4.2
异补骨脂素	2.18	4.36	8.71	96.1	95.8	99.7	2.9	4.1	2.0
8-甲氧基补骨脂素	2.09	4.18	8.37	86.7	88.9	93.5	2.4	2.8	1.7
欧前胡素	0.43	0.85	1.71	92.7	95.7	102.8	4.8	3.5	2.8
异欧前胡素	0.42	0.83	1.66	96.8	94.7	98.3	4.5	3.8	3.5
6，7-二甲氧基香豆素	0.42	0.84	1.68	87.4	90.8	98.5	3.6	2.9	3.4
新补骨脂异黄酮	0.42	0.84	1.67	93.4	91.5	96.2	4.7	3.7	2.8
补骨脂二氢黄酮	0.44	0.88	1.76	94.8	96.4	108.6	3.4	3.5	3.0
4-甲基-7-羟基香豆素	0.22	0.45	0.90	89.6	92.8	102.3	2.9	3.7	1.5
4-甲基-7-乙氧基香豆素	0.22	0.45	0.90	98.4	101.5	99.7	5.3	3.7	3.8
5-甲氧基补骨脂素	0.21	0.42	0.84	86.9	95.1	98.5	4.5	3.8	2.9
2，4，8-三甲基补骨脂素	0.20	0.41	0.81	92.8	102.7	101.4	4.7	3.2	2.4
环香豆素	0.20	0.40	0.81	92.4	96.5	98.1	5.4	3.7	3.0