高效液相色谱法同时测定化妆品中24种香豆素类化合物及质谱确证

doi:10.3969/j.issn.2097-2806.2024.01.016

摘要/Abstract

摘要：

采用高效液相色谱法，建立了同时测定化妆品中24种香豆素类化合物含量的分析方法。乳液、膏霜、水基类样品和唇膏、粉饼等固态类样品分别采用甲醇、四氢呋喃-甲醇（1∶1）混合溶液超声提取后，经色谱柱分离，以甲醇-0.02 mol/L磷酸二氢钠缓冲液（pH 3.5）为流动相进行梯度洗脱，流速1.0 mL/min，二极管阵列检测器多波长检测，外标法定量。经高效液相色谱法检出的疑似阳性样品，采用液相色谱-串联质谱法进行确证。结果表明，24种香豆素类化合物在各自的质量浓度范围内呈良好的线性关系，相关系数r均≥0.999 6，检出限为2~19 mg/kg，定量限为5~63 mg/kg。在低、中、高三个加标水平下，方法回收率为94.4%~106.4%，相对标准偏差为0.5%~3.3%。该方法操作简便、准确性好，可用于化妆品中香豆素类化合物的定性筛查和定量测定。

关键词: 香豆素, 化妆品, 高效液相色谱法, 液相色谱-串联质谱法

Abstract:

An analytical method for simultaneous determination of 24 coumarin compounds in cosmetics was established by high performance liquid chromatography. Samples with certain fluidity, such as lotions, creams, and water-based samples, were directly extracted with methanol by ultrasonic treatment, while solid samples （lipsticks, pressed powders, etc.） were ultrasonically extracted with mixed solvent of tetrahydrofuran-methanol （1∶1）. The compounds were separated on a CAPCELL CORE MGII C₁₈ （4.6×250 mm, 5 μm） column, and gradient elution was carried out with methanol and 0.02 mol/L sodium dihydrogen phosphate buffer （pH 3.5） as the mobile phase. The flow rate was 1.0 mL/min. The quantification was done by external standard method, with PDA detector at multi-wavelength, and the suspected positive samples were confirmed by liquid chromatography-tandem mass spectrometry. The results showed that the 24 coumarin compounds showed good linearity, with correlation coefficient≥0.999 6 in their respective mass concentration range. The limits of detection and limits of quantification were 2-19 mg/kg and 5-63 mg/kg, respectively. At the three spiked levels of low, medium and high, the average recoveries ranged from 94.4% to 106.4%, and the relative standard deviations were between 0.5% and 3.3%. This method was simple, accurate and suitable for the determination of 24 coumarin compounds in cosmetics.

Key words: coumarin, cosmetic, high performance liquid chromatography, liquid chromatography-tandem mass spectrometry

中图分类号:

TQ658

龙慧端, 鲁毅翔, 覃江兰, 张科明. 高效液相色谱法同时测定化妆品中24种香豆素类化合物及质谱确证[J]. 日用化学工业（中英文）, 2024, 54(1): 114-122.

Huiduan Long, Yixiang Lu, Jianglan Qin, Keming Zhang. Simultaneous determination of 24 coumarin compounds in cosmetics by high performance liquid chromatography and verification by liquid chromatography-tandem mass spectrometry[J]. China Surfactant Detergent & Cosmetics, 2024, 54(1): 114-122.

图/表 6

表1

表2

表3

图1

表4

表5

24种香豆素类化合物的加标回收率及精密度（n = 6）"

序号	名称	添加量/μg	乳液		霜膏		水基		固态基
序号	名称	添加量/μg	回收率/ %	RSD/ %	回收率/ %	RSD/ %	回收率/ %	RSD/ %	回收率/ %	RSD/ %
1	6-羟基-7-甲氧基香豆素	21.68	97.7	1.7	103.7	1.5	104.4	1.4	98.4	2.2
		43.36	97.9	2.2	105.3	2.3	102.1	1.6	101.1	1.8
		216.8	99.9	1.8	103.7	1.2	104.4	1.4	101.1	2.1
2	7-羟基-6-甲氧基香豆素	22.14	101.8	2.1	106.3	1.9	100.3	2.2	103.9	1.3
		44.28	105.6	2.0	99.4	1.9	107.4	3.0	102.3	2.3
		221.4	102.6	1.8	102.1	2.8	104.3	1.6	104.8	2.2
3	7-羟基香豆素	20.45	103.2	1.4	107.2	1.4	100.9	0.9	99.9	1.7
		40.90	100.4	1.2	106.8	1.9	101.8	2.1	102.4	2.4
		204.5	103.1	1.1	101.6	1.4	100.9	2.0	103.9	1.1
4	6，7-二甲氧基香豆素	30.60	98.5	2.7	105.6	1.1	100.7	0.9	99.9	1.7
		61.24	100.1	1.0	105.8	1.0	105.6	1.3	97.2	2.5
		306.0	96.9	2.3	101.8	1.8	100.7	0.9	101.1	2.1
5	香豆素	11.26	96.2	1.6	100.3	1.3	99.4	0.9	98.6	1.8
		22.52	101.6	0.8	101.8	1.9	102.6	1.2	100.4	2.2
		112.6	99.7	2.1	102.7	2.0	102.9	1.1	102.2	1.6
6	二氢香豆素	229.9	97.6	2.2	102.3	2.7	99.2	2.0	98.3	1.5
		459.7	100.9	1.7	103.7	2.5	104.5	3.3	105.3	2.5
		2 299	103.4	2.4	100.2	1.1	105.0	2.0	101.3	2.3
7	羟甲基香豆素	27.16	101.0	1.7	103.0	2.4	103.5	2.2	101.9	1.6
		54.32	97.3	0.9	108.2	1.1	98.8	2.4	102.4	2.7
		271.6	99.0	1.2	103.0	1.2	103.5	2.2	101.9	1.4
8	7-甲氧基香豆素	21.09	103.7	1.6	104.1	1.1	102.9	2.0	100.2	1.6
		42.18	100.4	1.9	97.6	0.9	101.4	1.7	101.7	1.9
		210.9	104.8	2.2	102.3	1.7	105.3	0.9	102.0	2.1
9	六氢香豆素	217.4	102.3	2.2	102.3	2.7	99.2	2.0	98.3	1.5
		434.8	99.2	2.0	102.0	2.2	102.8	1.5	98.4	1.9
		2 174	103.2	2.0	100.5	2.2	98.2	1.9	102.9	1.3
10	补骨脂素	22.14	107.2	1.0	102.2	1.9	100.4	1.6	105.3	0.8
		44.28	102.3	1.3	101.4	0.9	99.9	1.6	101.4	1.0
		221.4	103.3	1.4	102.6	2.0	101.7	2.0	100.9	1.4
11	7-甲基香豆素	23.07	104.1	2.0	103.6	1.1	104.4	1.6	102.6	2.0
		46.14	102.2	1.6	101.7	1.8	97.8	2.0	103.9	1.7
		230.7	99.6	1.8	100.6	0.9	101.3	1.6	104.4	1.8
12	6-甲基香豆素	31.84	101.2	1.2	100.3	1.6	101.2	1.5	100.4	0.9
		63.68	100.7	1.4	104.2	2.2	101.8	1.5	102.2	1.1
		318.4	98.6	1.9	102.5	2.1	103.3	2.1	99.3	1.4
13	异补骨脂素	13.34	103.4	2.3	99.5	2.4	103.9	1.9	102.5	2.0
		26.68	100.3	1.9	104.9	1.6	103.0	2.5	104.2	2.1
		133.4	101.3	2.3	98.5	1.6	103.6	1.4	101.7	2.4
14	8-甲氧基补骨脂素	31.88	102.6	1.0	99.7	1.1	104.6	1.4	103.6	2.0
		63.76	103.4	1.4	100.4	1.0	103.2	1.3	101.8	1.3
		318.8	101.8	1.6	102.6	1.9	101.8	2.6	100.5	2.4
15	7-甲氧基-5，11，12-三羟基香豆素	32.14	104.9	1.7	101.3	1.6	98.3	1.4	106.3	1.7
		64.28	103.0	0.9	98.6	1.6	103.2	1.2	102.6	1.9
		321.4	102.5	1.6	100.4	2.1	101.5	1.9	103.3	1.2
16	5-甲氧基补骨脂素	12.84	102.9	1.7	106.0	2.5	103.2	1.7	101.2	2.0
		25.67	97.9	1.1	105.5	1.5	102.1	1.6	103.7	3.4
		128.4	99.5	1.1	103.6	2.1	103.2	1.7	100.3	1.8
17	7-乙氧基-4-甲基香豆素	11.47	102.6	1.1	96.8	2.6	102.6	1.8	102.5	2.3
		22.93	97.8	1.1	102.0	2.1	103.3	1.4	103.7	1.1
		114.7	98.4	1.4	102.8	2.2	102.6	1.8	99.2	1.4
18	醋硝香豆素	11.29	99.2	2.2	106.2	3.5	102.7	2.0	103.0	2.9
		22.58	95.6	1.2	103.0	1.0	99.3	2.2	103.0	1.5
		112.9	99.0	1.8	103.5	2.5	102.7	2.0	100.2	1.8
19	双香豆素	15.48	104.3	2.4	100.3	2.1	103.3	2.1	102.5	1.5
		30.96	103.9	3.2	103.5	1.5	102.4	1.9	100.7	2.3
		154.8	95.8	1.9	101.2	2.3	104.3	2.5	105.4	2.1
20	苯丙香豆素	10.21	103.9	2.1	106.4	2.2	102.7	2.1	101.6	2.0
		20.42	99.6	2.4	105.2	1.1	103.5	2.3	101.6	2.5
		102.1	97.0	2.4	101.8	1.3	102.7	2.1	104.2	1.5
21	欧前胡素	10.33	99.3	3.2	99.0	2.5	98.7	2.8	101.5	2.3
		20.66	100.4	2.6	105.3	3.0	98.5	2.4	103.5	2.6
		103.3	98.0	1.1	96.8	1.5	98.7	2.8	99.4	3.3
22	三甲沙林	8.753	102.4	2.2	107.0	0.7	102.9	2.3	103.5	2.4
		17.51	103.0	2.7	107.5	1.7	103.5	2.3	102.4	1.4
		87.53	98.1	1.2	102.4	1.9	102.9	2.4	99.3	1.1
23	异欧前胡素	11.63	93.8	1.7	94.4	2.3	100.0	2.4	96.4	2.6
		23.27	97.1	2.1	93.0	1.4	98.4	2.1	95.8	2.0
		116.3	94.0	1.0	100.4	1.8	103.2	1.6	97.5	2.7
24	4，6-二甲基-8-特丁香豆素	21.96	102.9	1.5	98.0	2.6	102.7	1.7	102.5	1.5
		43.92	100.8	1.2	94.8	1.9	100.4	2.5	103.3	1.3
		219.6	98.4	1.7	102.3	1.3	102.7	1.7	99.8	0.9

表5

参考文献 23

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序号	名称	保留时间/min	母离子（m/z）	去簇电压/V	子离子（m/z）	碰撞电压/V
1	6-羟基-7-甲氧基香豆素	6.57	193.0	80	178.1^*/132.8	29/26
2	7-羟基-6-甲氧基香豆素	6.46	193.0	90	133.0^*/150.1	23/36
3	7-羟基香豆素	7.12	163.0	80	106.9^*/ 90.8	29/26
4	6，7-二甲氧基香豆素	8.54	207.1	80	151.1^*/163.1	28/31
5	香豆素²	8.68	147.1	60	90.9^*/102.8	28/22
6	二氢香豆素¹	11.28	149.0	120	107.0^*/ 79.1	23/32
7	羟甲基香豆素	8.78	176.9	100	77.1^*/105.0	48/26
8	7-甲氧基香豆素¹	10.40	177.1	80	121.1^*/132.9	29/24
9	六氢香豆素¹	12.46	153.0	65	93.1^*/ 77.2	18/40
10	补骨脂素	10.91	187.1	160	131.0^*/115.1	33/29
11	7-甲基香豆素¹	11.87	161.1	80	104.9^*/ 91.0	29/30
12	6-甲基香豆素	12.02	161.1	80	104.9^*/ 91.0	28/27
13	异补骨脂素	12.30	187.1	160	131.0^*/115.1	30/29
14	8-甲氧基骨补脂素	11.87	217.0	90	201.9^*/174.1	27/37
15	7-甲氧基-5，11，12-三羟基香豆素	13.66	315.1	120	161.0^*/297.0	48/32
16	5-甲氧基骨补脂素	14.49	217.0	80	201.9^*/174.1	29/37
17	7-乙氧基-4-甲基香豆素¹	16.26	205.1	80	177.0^*/120.8	25/34
18	醋硝香豆素¹	17.82	354.2	100	162.9^*/295.9	22/27
19	双香豆素¹	18.86	335.1	-20	160.9^*/117.0	-18/-56
20	苯丙香豆素¹	19.12	281.1	100	203.2^*/174.9	24/33
21	欧前胡素¹	18.73	271.0	105	203.1^*/147.0	16/42
22	三甲沙林	19.60	229.1	120	142.0^*/173.0	30/30
23	异欧前胡素	20.02	271.0	80	203.1^*/147.0	14/41
24	4，6-二甲基-8-特丁香豆素¹	20.29	231.0	100	215.0^*/174.8	42/27

序号	名称	线性范围/（mg/L）	回归方程	相关系数	LOD/（mg/kg）	LOQ/（mg/kg）
1	6-羟基-7-甲氧基香豆素	2~160	Y=18 799X-5 482	0.999 8	7	21
2	7-羟基-6-甲氧基香豆素	2~160	Y=18 556X-1 895	0.999 9	6	18
3	7-羟基香豆素	2~160	Y=31 472X-5 022	0.999 9	6	18
4	6，7-二甲氧基香豆素	3~240	Y=12 707X+19 892	0.999 7	9	30
5	香豆素	1~80	Y=29 937X-4 469	0.999 9	3	9
6	二氢香豆素	20~1 600	Y=19 310X-7 654	0.999 7	67	209
7	羟甲基香豆素	2~160	Y=30 102X-12 972	0.999 8	7	23
8	7-甲氧基香豆素	2~160	Y=31 525X-24 107	0.999 7	7	23
9	六氢香豆素	20~1 600	Y=4 807X-524	0.999 7	71	224
10	补骨脂素	2~160	Y=27 116X-5 282	0.999 9	7	21
11	7-甲基香豆素	2~160	Y=37 836X-32 702	0.999 8	8	27
12	6-甲基香豆素	3~240	Y=19 722X-11 963	0.999 8	11	33
13	异补骨脂素	1~80	Y=29 500X-586	0.999 9	3	9
14	8-甲氧基补骨脂素	3~240	Y=34 568X-31 916	0.999 7	10	33
15	7-甲氧基-5，11，12-三羟基香豆素	3~240	Y=31 655X-12 251	0.999 9	11	33
16	5-甲氧基补骨脂素	1~80	Y=32 955X-2 520	0.999 9	3	9
17	7-乙氧基-4-甲基香豆素	1~80	Y=27 002X-514	0.999 9	3	9
18	醋硝香豆素	1~80	Y=29 121X-1 254	0.999 8	3	8
19	双香豆素	1.5~120	Y=12 414X-3 461	0.999 7	5	16
20	苯丙香豆素	1~80	Y=29 348X+2 129	0.999 9	3	8
21	欧前胡素	1~80	Y=29 343X-149	0.999 8	3	8
22	三甲沙林	1~80	Y=31 472X-12 188	0.999 7	3	9
23	异欧前胡素	1~80	Y=27 831X+694	0.999 9	3	9
24	4，6-二甲基-8-特丁香豆素	2~160	Y=12 145X-2 186	0.999 9	6	19