超高效液相色谱-串联质谱法测定化妆品中36种兴奋剂

doi:10.3969/j.issn.2097-2806.2024.03.016

摘要/Abstract

摘要：

利用超高效液相色谱-串联质谱仪（UPLC-MS/MS），建立了一种同时测定化妆品中蛋白同化制剂、β2激动剂、糖皮质激素、刺激剂、大麻酚类等36种兴奋剂的检测方法。化妆品样品经甲醇超声提取后，以0.1%乙酸水溶液和甲醇-乙腈混合溶液（体积比1∶9）作为流动相，使用Waters ACQUITY UPLC BEH C18色谱柱（3.0 mm×150 mm，1.7 μm）进行梯度分离。目标物在电喷雾正负离子模式下，以多反应监测（MRM）方式进行测定，用外标法定量。结果表明，在1.0~20 ng/mL的质量浓度范围内，36种兴奋剂组分的线性关系良好（r>0.995），检出限为1.0~3.0 μg/kg，定量限为3.3~9.9 μg/kg。3种基质样品在低、中、高3个加标水平下的平均回收率为80.6%~115.3%，相对标准偏差为0.5%~12.9% （n=5）。该方法前处理操作简便，快速，灵敏度高，结果准确，适用于常见化妆品中多类兴奋剂组分的快速检测。

关键词: 超高效液相色谱-串联质谱法, 兴奋剂, 化妆品

Abstract:

An ultra performance liquid chromatography-tandem mass spectrometric （UPLC-MS/MS） method was developed for the simultaneous determination of 36 stimulants including anabolic agents, β2 agonists, glucocorticoids, irritants, and cannabinoids in cosmetics. In this study, pretreatment operation and instruments parameters were also optimized. Cosmetic samples were ultrasonically extracted by methanol, and then the separation of 36 stimulants was carried out on a Waters ACQUITY UPLC BEH C18 column （3.0 mm×150 mm, 1.7 μm） by gradient elution, using 0.1% acetic acid solution and methanol-acetonitrile （1∶9, by volume） as the mobile phase. The 36 target compounds were analyzed in positive and negative electrospray ionization mode under multiple reaction monitoring （MRM） mode. Meanwhile, the analytes were quantified by external standard method, and the linear regression analysis data show that there are good linear relationships for 36 target compounds in the mass concentration range of 1.0-20 ng/mL, with their correlation coefficients （r） greater than 0.995. The limits of detection （LOD） and limits of quantitation （LOQ） of the method are in the ranges of 1.0-3.0 μg/kg and 3.3-9.9 μg/kg, respectively. The recoveries for 36 target compounds in three kinds of matrix samples at three spiked levels are between 80.6% and 115.3%, with the relative standard deviations （RSD） of 0.5%-12.9% （n=5）. The method is simple, rapid, high sensitivity, accurate and suitable for the rapid detection of many stimulants in cosmetics.

Key words: ultra performance liquid chromatography-tandem mass spectrometry, stimulant, cosmetics

中图分类号:

TQ658

徐炜, 邹坡, 李长于, 杨铭, 鹿燕, 李慧良. 超高效液相色谱-串联质谱法测定化妆品中36种兴奋剂[J]. 日用化学工业（中英文）, 2024, 54(3): 360-368.

Wei Xu, Po Zou, Changyu Li, Ming Yang, Yan Lu, Huiliang Li. Determination of 36 stimulants in cosmetics by ultra performance liquid chromatography-tandem mass spectrometry[J]. China Surfactant Detergent & Cosmetics, 2024, 54(3): 360-368.

图/表 7

表1

36种兴奋剂的保留时间及质谱参数"

序号	化合物名称	保留时间/min	电离方式	母离子（m/z）	子离子（m/z）	去簇电压/V	碰撞电压/V
1	沙丁胺醇（Salbutamol）	5.86	ESI+	240.2	148.2^*/166.2	78	26/19
2	丙卡特罗（Procaterol）	6.42	ESI+	291.2	273.3^*/231.1	81	21/29
3	去甲乌药碱（Higenamine）	6.74	ESI+	272.1	107.0^*/255.1	50	31/20
4	麻黄碱（Ephedrine）	7.27	ESI+	166.0	117.0^*/148.3	84	26/17
5	左去氧麻黄碱（Levmetamfetamine）	8.61	ESI+	150.1	91.1^*/119.1	70	23/17
6	士的宁（Strychnine）	8.83	ESI+	335.2	184.2^*/156.2	90	50/60
7	克仑特罗（Clenbuterol）	10.75	ESI+	277.0	203.0^*/132.0	51	23/37
8	妥洛特罗（Tulobuterol）	10.79	ESI+	228.0	154.0^*/119.0	85	23/39
9	海洛因（Heroin）	11.09	ESI+	370.2	268.2^*/165.0	90	38/60
10	可卡因（Cocaine）	11.69	ESI+	304.2	182.3^*/150.1	60	28/35
11	泼尼松（Prednisone）	14.03	ESI+	359.2	147.2^*/341.2	80	35/15
12	泼尼松龙（Prednisolone）	14.07	ESI+	371.2	343.2^*/147.2	80	14/34
13	氢化可的松（Hydrocortisone）	14.24	ESI+	373.2	121.1^*/105.0	80	31/68
14	可的松（Cortisone）	14.28	ESI+	371.2	163.2^*/121.1	80	34/47
15	甲基泼尼松龙（Methylprednisolone）	15.58	ESI+	375.2	339.2^*/161.2	66	14/28
16	倍他米松（Betamethasone）	15.70	ESI+	393.2	355.2^*/337.3	80	15/19
17	地塞米松（Dexamethasone）	15.84	ESI+	393.2	373.4^*/355.2	80	15/15
18	倍氯米松（Beclomethasone）	16.17	ESI+	409.2	391.2^*/279.3	80	15/29
19	氟甲睾酮（Fluoxymesterone）	16.21	ESI+	337.2	241.2^*/131.2	160	35/42
20	群勃龙（Trenbolone）	16.46	ESI+	271.2	253.2^*/199.2	80	26/32
21	曲安奈德（Triamcinolone acetonide）	16.48	ESI+	435.2	339.2^*/397.2	80	15/15
22	氟尼缩松（Flunisolide）	16.69	ESI+	435.2	339.1^*/321.1	80	18/19
23	地夫可特（Deflazacort）	17.78	ESI+	442.3	124.1^*/142.1	80	65/45
24	美雄酮（Metandienone）	17.81	ESI+	301.2	121.1^*/149.2	100	28/23
25	沙美特罗（Salmeterol）	17.82	ESI+	416.2	398.2^*/380.1	100	20/26
26	睾酮（Testosterone）	18.34	ESI+	289.2	97.1^*/109.0	113	29/30
27	孕三烯酮（Gestrinone）	18.97	ESI+	309.1	241.1^*/199.2	120	29/43
28	布地奈德（Budesonide）	19.03	ESI+	431.2	413.2^*/147.1	80	15/42
29	甲睾酮（Methyltestosterone）	19.28	ESI+	303.2	108.9^*/97.0	110	30/29
30	螺内酯（Spironolactone）	19.70	ESI+	341.2	107.2^*/91.0	120	45/79
31	坎利酮（Canrenone）	19.87	ESI+	341.2	107.2^*/91.0	120	45/79
32	雄酮（Androsterone）	22.10	ESI+	291.2	273.3^*/255.0	70	13/21
33	大麻二酚（Cannabidiol）	29.06	ESI-	313.1	245.1^*/179.1	-100	-30/-25
34	环索奈德（Ciclesonide）	30.88	ESI+	541.2	323.2^*/147.0	80	23/35
35	大麻酚（Cannabinol）	30.97	ESI-	309.1	279.1^*/222.1	-120	-40/-55
36	Δ⁹-四氢大麻酚（Δ⁹-tetrahydrocannabinol）	31.90	ESI-	313.1	245.1^*/191.1	-130	-34/-34

表1

图1

图2

图3

表2

36种兴奋剂的线性回归方程、相关系数、检出限和定量限"

序号	化合物名称	线性方程	r	检出限/（μg/kg）	定量限/（μg/kg）
1	沙丁胺醇	y=182 701 x+34 609.0	0.999 0	1.0	3.3
2	丙卡特罗	y=65 487 x+13 728.5	0.998 9	2.8	9.3
3	去甲乌药碱	y=15 446 x-5 724.6	0.999 8	1.0	3.3
4	麻黄碱	y=30 735 x+8 600.2	0.998 7	1.0	3.3
5	左去氧麻黄碱	y=361 921 x+94 797.6	0.998 1	1.0	3.3
6	士的宁	y=46 669 x+10 620.5	0.999 6	1.0	3.3
7	克仑特罗	y=312 267 x+61 973.0	0.998 4	1.0	3.3
8	妥洛特罗	y=346 559 x+74 408.5	0.999 2	1.0	3.3
9	海洛因	y=28 115 x+3 889.7	0.999 9	1.1	3.6
10	可卡因	y=534 736 x+102 549.0	0.999 5	1.7	5.7
11	泼尼松	y=22 469 x+5 151.9	0.998 3	1.0	3.4
12	泼尼松龙	y=112 145 x+25 809.5	0.999 0	2.0	6.7
13	氢化可的松	y=31 039 x+6 999.1	0.999 2	2.0	6.5
14	可的松	y=46 518 x+12 035.4	0.999 3	2.1	7.0
15	甲基泼尼松龙	y=51 899 x+8 369.7	0.999 8	1.4	4.8
16	倍他米松	y=55 382 x+9 860.9	0.999 0	1.0	3.3
17	地塞米松	y=86 958 x+17 910.5	0.998 4	1.0	3.3
18	倍氯米松	y=147 290 x+38 076.2	0.998 9	1.0	3.3
19	氟甲睾酮	y=24 862 x+6 182.6	0.999 2	1.0	3.4
20	群勃龙	y=83 150 x+26 785.4	0.996 9	1.0	3.3
21	曲安奈德	y=26 502 x+7 257.9	0.996 9	1.0	3.3
22	氟尼缩松	y=47 515 x+10 044.9	0.998 1	1.0	3.3
23	地夫可特	y=20 043 x+5 025.8	0.999 5	1.0	3.3
24	美雄酮	y=172 129 x+46 234.9	0.998 9	1.0	3.3
25	沙美特罗	y=258 896 x+28 276.8	0.999 0	1.0	3.3
26	睾酮	y=138 465 x+21 686.9	0.999 1	1.0	3.3
27	孕三烯酮	y=82 263 x+18 661.3	0.998 3	1.0	3.3
28	布地奈德	y=98 681 x+28 670.2	0.998 0	2.9	9.7
29	甲睾酮	y=65 812 x+15 059.5	0.998 1	1.7	5.5
30	螺内酯	y=24 197 x+5 817.9	0.999 5	2.9	9.7
31	坎利酮	y=43 612 x+6 307.8	0.999 8	1.0	3.3
32	雄酮	y=52 912 x+12 100.1	0.998 9	3.0	9.9
33	大麻二酚	y=85 066 x+1 469.1	0.998 5	1.0	3.3
34	环索奈德	y=38 549 x+12 054.4	0.996 6	2.5	8.3
35	大麻酚	y=86 412 x+11 753.7	0.996 8	1.0	3.3
36	Δ⁹-四氢大麻酚	y=9 957 x+1 878.3	0.995 6	1.0	3.3

表2

表3

36种兴奋剂的回收率和精密度（n=5）"

序号	化合物名称	水剂		膏霜		蜡基
序号	化合物名称	回收率/%	RSD/%	回收率/%	RSD/%	回收率/%	RSD/%
1	沙丁胺醇	86.3~100.0	0.9~8.9	83.8~85.6	1.5~2.0	84.6~92.6	0.9~2.4
2	丙卡特罗	86.5~101.1	0.7~7.8	86.8~95.6	2.8~6.3	85.1~93.9	2.9~4.4
3	去甲乌药碱	83.6~103.4	3.0~12.9	81.3~88.1	4.2~12.5	80.9~88.9	2.9~3.4
4	麻黄碱	88.7~98.9	1.6~8.2	85.3~89.1	0.5~2.8	89.5~95.2	1.0~1.8
5	左去氧麻黄碱	96.1~101.2	2.1~3.2	84.6~105.0	1.4~3.7	93.5~100.3	0.9~2.0
6	士的宁	87.7~100.4	1.7~8.9	86.7~95.0	1.2~3.2	91.1~105.7	0.8~3.1
7	克仑特罗	100.9~106.1	1.3~4.4	84.3~89.5	1.1~2.6	92.1~98.8	1.1~1.7
8	妥洛特罗	99.0~102.4	1.1~3.6	81.3~86.0	1.8~3.1	92.5~99.2	0.8~2.0
9	海洛因	88.8~95.2	1.1~3.9	86.5~95.2	0.9~4.1	86.4~102.6	1.8~5.7
10	可卡因	95.4~101.4	1.2~4.9	86.6~90.2	0.7~2.2	95.5~99.7	0.5~1.7
11	泼尼松	94.3~104.1	1.3~3.6	86.7~110.5	2.3~3.6	94.3~105.6	2.5~4.7
12	泼尼松龙	94.8~102.1	1.6~4.6	85.3~104.7	1.5~2.3	94.8~98.4	1.7~2.7
13	氢化可的松	94.3~104.6	3.4~6.4	86.9~106.1	2.2~3.2	93.1~93.6	1.8~3.7
14	可的松	94.8~101.7	1.8~3.7	90.4~105.9	0.9~3.8	89.6~102.2	3.3~3.6
15	甲基泼尼松龙	93.2~100.6	2.4~4.6	87.0~109.7	1.1~2.9	93.1~99.6	1.5~2.3
16	倍他米松	97.0~99.7	1.6~3.0	83.8~111.6	1.6~4.6	92.8~101.6	1.2~2.3
17	地塞米松	92.9~100.1	2.4~3.5	83.5~104.8	1.2~4.3	94.6~98.8	1.6~2.3
18	倍氯米松	94.7~98.9	1.2~3.3	86.5~109.3	1.7~2.7	92.7~101.9	0.7~2.5
19	氟甲睾酮	87.3~100.2	3.8~4.6	87.6~102.4	3.9~8.1	93.1~96.6	2.2~4.5
20	群勃龙	93.8~103.9	1.4~2.5	86.0~106.0	2.1~4.8	95.9~98.7	1.1~2.8
21	曲安奈德	96.2~106.3	2.8~3.0	88.9~111.9	2.5~5.6	94.8~103.9	0.5~4.2
22	氟尼缩松	97.1~104.7	2.6~4.6	82.8~108.8	2.8~4.9	88.1~97.4	2.0~4.7
23	地夫可特	91.6~95.7	3.6~4.8	86.5~103.9	3.5~7.1	90.7~97.6	1.7~3.4
24	美雄酮	88.5~102.5	1.4~4.0	86.6~111.3	2.6~3.9	91.5~97.0	1.8~2.9
25	沙美特罗	95.1~102.8	0.7~5.9	90.2~107.3	2.1~3.4	93.1~100.9	1.5~2.1
26	睾酮	91.8~100.2	2.0~2.6	90.0~111.7	1.6~2.7	93.5~98.1	1.8~2.9
27	孕三烯酮	93.9~103.5	1.9~4.0	82.6~90.1	2.4~4.9	94.8~98.2	2.3~3.2
28	布地奈德	88.7~101.5	1.4~4.0	83.5~96.1	2.3~4.2	91.4~100.1	2.2~3.7
29	甲睾酮	89.4~99.3	3.2~5.2	89.7~106.5	3.3~5.0	92.9~101.5	3.7~5.9
30	螺内酯	90.3~98.8	1.7~6.1	103.5-114.4	2.1~7.5	85.6~95.2	4.0~5.6
31	坎利酮	90.2~104.3	1.2~4.0	94.6~100.6	1.6~2.7	88.1~96.9	3.7~5.2
32	雄酮	94.1~114.7	1.9~5.7	103.6~111.3	3.2~10.6	80.9~87.4	3.9~5.1
33	大麻二酚	80.6~99.3	2.9~5.0	90.1~113.9	1.3~5.5	93.1~96.8	1.7~2.7
34	环索奈德	100.0~101.7	2.2~4.8	88.9~100.5	2.5~7.3	92.2~102.5	1.5~3.0
35	大麻酚	86.4~106.4	3.0~8.7	97.4~115.3	2.9~11.7	84.3~95.6	2.4~5.7
36	Δ⁹-四氢大麻酚	90.9~102.6	4.7~9.2	94.0~114.1	3.6~10.6	88.1~98.4	2.4~6.8

表3

图4

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