GC-MS和LC-MS/MS法测定毛发中的6种苯丙胺类药物

韦棋; 苏福海

doi:10.12338/j.issn.2096-9015.2021.0611

GC-MS和LC-MS/MS法测定毛发中的6种苯丙胺类药物

doi: 10.12338/j.issn.2096-9015.2021.0611

韦棋,
苏福海^,

中国计量科学研究院，北京 100029

基金项目: 国家重点研发计划( 2018YFC0807301-4) 。

详细信息

作者简介:
韦棋( 1995-)，中国计量科学研究院硕士研究生，研究方向：药物分析，邮箱：weiqi18811@163.com

通讯作者:
苏福海( 1976-) ，中国计量科学研究院副研究员，研究方向：有机分析，邮箱：sufh@nim.ac.cn

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出版历程
- 录用日期: 2022-03-31
- 网络出版日期: 2022-04-12

The Determination of 6 Amphetamine Drugs in Hair Samples by GC-MS and LC-MS/MS

WEI Qi,
SU Fuhai^,

National Institute of Metrology, Beijing 100029, China

摘要

摘要: 建立毛发样本中6种苯丙胺类药物的GC-MS和LC-MS/MS分析检测方法。选择合适的人体毛发，浸泡在药物-二甲基亚砜溶液中制备含6种苯丙胺类药物的毛发样本，采用单因素变量法优化样品前处理过程，利用GC-MS和LC-MS/MS法对制备的毛发样本进行定量分析。以甲基苯丙胺-D5为内标，采用工作曲线法进行定量分析。实验结果显示，制备的毛发样本中6种苯丙胺类药物的最佳前处理方法为：提取溶剂0.01 mol·L⁻¹ HCl，液料比100:1（mL·g⁻¹），提取温度40 ℃，提取时间50 min。GC-MS中，6种苯丙胺类药物在0.5～5.0 ng·mg⁻¹浓度范围内线性关系良好，r²>0.999，检测限为0.04～0.08 ng·mg⁻¹，回收率≥85.46%，RSDs≤ 3.6%（n=9）；LC-MS/MS中，6种苯丙胺类药物在0.06～0.50 ng·mg⁻¹浓度范围内线性关系良好，r²>0.999，检测限低至0.05～5.0 pg·mg⁻¹，回收率在86.50%～111.9%范围内，RSDs≤ 4.2%（n=9）。这两种方法应用于毛发中苯丙胺类药物的检测，灵敏度高、准确度好、专属性强，可作为毛发中苯丙胺类药物基体标准物质的定值方法，用于标准物质的研制，应用于法医滥用药物检测和实验室质量控制领域。
- 苯丙胺 /
- 毛发样本 /
- 标准物质 /
- 气相色谱串联质谱法 /
- 液相色谱串联质谱法
Abstract: A method was developed for the analytical determination of six amphetamine drugs in hair samples by GC-MS and LC-MS/MS. The hair samples were prepared by immersing in the drug-dimethyl sulfoxide solution including target analytes. The sample pretreatment process was optimized using the one-way variable method, and the prepared hair samples were quantified using GC-MS and LC-MS/MS methods. The working curve method was used for quantitative analysis with methamphetamine-D5 as the internal standard. The results showed that the optimal pretreatment methods for the six amphetamine drugs in the prepared hair samples were: 0.01 mol·L⁻¹ HCl, liquid-material ratio 100:1 (mL·g⁻¹), extraction temperature 40 ℃, extraction time 50 min. In GC-MS, the linear range of 6 amphetamine drugs was 0.5 ~ 5.0 ng·mg⁻¹, r²>0.999. The detection limits of the method were 0.04～0.08 ng·mg⁻¹, and the recovery rates were greater than 85.46%, RSDs≤3.6% (n=9). In LC-MS/MS, the 6 amphetamine drugs showed a good linear relationship within the concentration range of 0.06～0.50 ng·mg⁻¹, r²>0.999. The detection limits were 0.05～5.0 pg·mg⁻¹and the recovery of appended contrast in the blank sample ranged from 86.50% to 111.9%, RSDs≤4.2% (n=9). The established methods were used for the detection of amphetamine drugs in human hair, with high sensitivity, good accuracy, and strong specificity. These two methods can be used as the certification method of the reference material of the amphetamine drug in human hair, which could be applied for drug abuse testing and laboratory quality control in the forensic field.
- amphetamine /
- hair samples /
- reference material /
- GC-MS /
- LC-MS/MS

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图 1 不同提取溶剂对苯丙胺类药物浓度的影响

Figure 1. The effect of different extraction solvents on the concentration of amphetamines

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图 2 不同提取温度对药物浓度的影响

Figure 2. The effect of different extraction temperature on drug concentration

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图 3 不同提取时间对药物浓度的影响

Figure 3. The effect of different extraction time on drug concentration

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图 4 不同液料比对药物浓度的影响

Figure 4. The effect of different liquid-to-material ratios on drug concentration

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图 5 不同盐酸浓度对药物浓度的影响

Figure 5. The effect of different hydrochloric acid concentration on drug concentration

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图 6 头发样本中6种苯丙胺类药物MRM色谱图

Figure 6. MRM chromatograms of 6 amphetamine drugs in hair samples

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图 7 头发样本中6种苯丙胺类药物的含量

Figure 7. The levels of 6 amphetamine drugs in hair samples

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表 1 实验仪器信息表

Table 1. Experimental instrument information sheet

仪器名称	型号	生产厂家
液质联用仪	TQS	Waters，美国
气质联用仪	7890A-7000	Agilent，美国
超声波清洗机	Branson 8510	Danbury，美国
氮吹仪	N-EVAP 112	Organomation，美国
超纯水机	Milli-Q	Millipore，美国
电热恒温鼓风干燥箱	DGG-9123AD	上海森信实验仪器有限公司
多用途冷冻离心机	CF16RXII	HITACHI公司
电子天平	XP205	Mettler Toledo公司，瑞士

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表 2 实验试剂信息表

Table 2. Laboratory reagent information sheet

试剂名称	纯度	生产厂家
甲醇	色谱纯	Merck KGaA，德国
乙腈	色谱纯	Merck KGaA，德国
盐酸	分析纯	北京化学试剂研究所有限责任公司，中国
二甲基亚砜	>99.5%	Honeywell，美国
甲酸	色谱纯	Honeywell，美国
乙酸铵	色谱纯	Honeywell，美国

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表 3 GC-MS中化合物的保留时间和特征离子

Table 3. Retention time and characteristic ions of amphetamine compounds in GC-MS

药物名称	SIM离子	保留时间（min）		RSD （n=3, %）
药物名称	SIM离子	标准溶液	头发样本	RSD （n=3, %）
AP	44*,91,117,65	5.69	5.70	0.07
MA	58*,91,56	6.45	6.46	0.09
MDEA	72*,135,136,77	11.60	11.60	0.02
MDMA	58*,77,59,135	11.10	11.10	0.01
MDA	44*,136,77,51	10.40	10.40	0.06
麻黄碱	58*,77,105	9.03	9.03	0.02
甲基苯丙胺-D5	62*,66,92,139	6.41	6.41	0.02
注：*为定量离子

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表 4 LC-MS/MS梯度洗脱程序表

Table 4. LC-MS/MS gradient elution program table

时间（min)	流动相A（%）	流动相B（%）
0	85	15
4	72	28
5	72	28
10	70	30
13	55	45
13.5	5	95
14.5	85	15
20	85	15

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表 5 LC-MS/MS中6种苯丙胺类化合物质谱参数表

Table 5. Mass spectrometry parameters of 6 amphetamine compounds in LC-MS/MS

药物名称	分子量	母离子 m/z	子离子 m/z	锥孔电压（V）	CE (V)	内标
AP	135.00	136.00	91.00*	55	25	甲基苯丙胺-D5
AP	135.00	136.00	119.00	55	15
MA	149.23	148.24	133.24*	64	18
MA	149.23	148.24	117.26	64	16
MDEA	207.00	208.22	105.23*	28	22
MDEA	207.00	208.22	132.85	28	24
MDMA	194.00	194.22	163.27*	32	12
MDMA	194.00	194.22	105.23	32	22
MDA	179.20	180.00	135.00	70	26
MDA	179.20	180.00	163.00*	70	13
麻黄碱	148.00	148.16	114.91	42	16
麻黄碱	148.00	148.16	133.24*	42	16
注：*为定量离子

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表 6 6种苯丙胺类化合物的检测限和定量限

Table 6. LOD and LOQ of 6 amphetamine compounds

药物名称	LOD（ng·mg⁻¹)		LOQ（ng·mg⁻¹)
药物名称	GC-MS	LC-MS	GC-MS	LC-MS
AP	0.04	0.005	0.08	0.02
MA	0.04	0.00005	0.08	0.0003
MDEA	0.04	0.00005	0.08	0.0003
MDMA	0.04	0.00005	0.08	0.0003
MDA	0.04	0.005	0.08	0.01
麻黄碱	0.08	0.00005	0.1	0.0003

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表 7 GC-MS中6种苯丙胺类化合物的线性与范围

Table 7. Linearity and range of 6 amphetamine compounds in GC-MS

药物名称	线性范围(ng·mg⁻¹)	回归方程	r²
AP	0.5～5.0	y=0.5272x−0.0009	0.9995
MA	0.5～5.0	y=0.206x+0.0358	0.9992
MDEA	0.5～5.0	y=0.6042x−0.0248	0.9993
MDMA	0.5～5.0	y=0.5188x−0.011	0.9997
MDA	0.5～5.0	y=0.2284x−0.0275	0.9998
麻黄碱	0.5～5.0	y=0.2303x−0.1461	0.9990

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表 8 LC-MS/MS种6种苯丙胺类化合物的线性与范围

Table 8. Linearity and range of 6 amphetamine compounds in LC-MS/MS

药物名称	线性范围(ng·mg⁻¹)	回归方程	r²
AP	0.06～0.50	y=0.1779x+0.0163	0.9991
MA	0.06～0.50	y=4.5592x+0.1073	0.9992
MDEA	0.06～0.50	y=10.369x+0.2026	0.9992
MDMA	0.06～0.50	y=15.063x+0.4445	0.9990
MDA	0.06～0.50	y=0.0566x+0.0062	0.9992
麻黄碱	0.06～0.50	y=5.4438x+0.1192	0.9998

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表 9 6种苯丙胺类化合物重复性实验结果（n=6）

Table 9. Results of reproducible experiments of amphetamine compounds（n=6）

药物名称	RSD (%)
药物名称	GC-MS	LC-MS/MS
AP	0.68	2.5
MA	1.1	1.0
MDEA	1.9	1.2
MDMA	0.32	0.75
MDA	1.9	2.7
麻黄碱	2	0.41

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表 10 苯丙胺类化合物的空白加标回收率（GC-MS）

Table 10. Results of blank spike recovery of amphetamine compounds（GC-MS）

药物名称	添加浓度/ng·mg⁻¹	测得浓度/ng·mg⁻¹	回收率/%	RSD/%
AP	0.8776	0.9240	105.3	1.5
	1.564	1.796	114.8	1.0
	3.048	3.090	101.4	2.6
MA	0.9900	0.8583	86.69	2.0
	1.764	1.691	95.86	1.6
	3.438	3.760	109.4	1.6
MDEA	0.9853	0.8421	85.46	0.66
	1.756	1.586	90.31	0.72
	3.422	3.163	92.44	2.4
MDMA	0.9814	0.8857	90.25	3.6
	1.749	1.798	102.8	0.21
	3.408	3.403	99.85	3.6
MDA	1.004	0.8742	87.10	0.78
	1.789	2.124	118.7	1.5
	3.486	4.122	118.3	1.7
麻黄碱	0.9022	0.9246	102.5	1.2
	1.608	1.706	106.1	2.0
	3.133	2.872	91.66	0.48

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表 11 苯丙胺类化合物的空白加标回收率（LC-MS/MS）

Table 11. Results of blank spike recovery of amphetamine compounds（LC-MS/MS）

药物名称	添加浓度/ng·mg⁻¹	测得浓度/ng·mg⁻¹	回收率/%	RSD/%
AP	0.07370	0.07916	107.4	4.2
	0.1750	0.1514	86.50	2.0
	0.3487	0.3207	91.96	2.6
MA	0.06520	0.05649	86.64	1.1
	0.1792	0.1840	102.7	1.0
	0.3569	0.3604	101.0	1.7
MDEA	0.06421	0.05989	93.27	1.8
	0.1764	0.1899	107.6	0.37
	0.3515	0.3662	104.2	1.0
MDMA	0.06397	0.05565	87.00	2.5
	0.1758	0.1967	111.9	1.4
	0.3502	0.3545	101.2	1.5
MDA	0.06283	0.06698	106.6	0.53
	0.1726	0.1525	88.32	1.2
	0.3439	0.3228	93.87	1.6
麻黄碱	0.06251	0.05480	87.67	1.6
	0.1718	0.1747	101.7	1.4
	0.3421	0.3492	102.1	0.69

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