液相色谱-四极杆/飞行时间质谱法快速筛查测定药物中芬太尼类物质

doi:10.3724/SP.J.1123.2019.11010

摘要/Abstract

摘要：

基于液相色谱-四极杆/飞行时间质谱（LC-QTOF-MS）建立了药物中27种新型毒品芬太尼类物质（芬太尼及其类似物）的同时快速筛查、确证和定量分析方法。样品经5 mL 75%（v/v）乙腈水溶液和5 mL乙腈提取，HLB固相萃取柱净化，电喷雾电离，正离子扫描，TOF-MS和信息依赖性采集（IDA）-MS/MS采集模式检测，外标法定量；建立一级精确质量数据库、保留时间和二级质谱库，实现了固体压片、粉末、液体药物及透皮贴剂中27种芬太尼类物质的快速筛查和确证。在5.00~100 μg/L范围内，27种目标化合物的线性关系良好（r²>0.99）。27种化合物的定量限均为10.0 μg/kg。在维生素C压片、头痛粉、止咳水、透皮贴剂样品中3个添加水平的平均回收率范围分别82.9%~106%、84.8%~106%、86.9%~109%和83.1%~106%，相对标准偏差（RSD）为0.38%~8.71%（n=6）。本方法操作简便，耗时短，灵敏度高，稳定性好，适用于液体和固体药物中芬太尼类物质的筛查检测及定量分析，可显著降低检测成本，具有实际应用价值。

关键词: 液相色谱-四极杆/飞行时间质谱, 高分辨质谱, 芬太尼类物质, 药物, 筛查

Abstract:

A method based on liquid chromatography coupled with high-resolution quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) was developed for the simultaneous screening and determination of fentanyl and its 26 analogs in liquid and solid powder drugs. The established method involves successive extraction by 5 mL 75% (v/v) acetonitrile aqueous solution and 5 mL acetonitrile, followed by clean-up using the hydrophilic lipophilic balance (HLB) solid-phase extraction method. Detection was achieved by electrospray ionization (ESI) in the positive mode, TOF-MS, and information-dependent acquisition (IDA)-MS/MS acquisition; the external standard method was adopted for quantification. Two databases of accurate mass and fragment ions were created. The standard matrix-matched calibration curves of the 27 target compounds were linear in the range of 5.00-100 μg/L, with the correlation coefficients (r²)>0. 99. The limits of quantification for the 27 target compounds were 10.0 μg/kg. The recoveries for all the target compounds in vitamin C tablet, headache powder, cough syrup, and transdermal patch samples were in the ranges of 82.9%-106%, 84.8%-106%, 86.9%-109%, and 83.1%-106%, respectively, with relative standard deviations ranging from 0.38% to 8.71% (n=6). The results demonstrated that the developed method is rapid and sensitive for the simultaneous monitoring and determination of fentanyl and 26 its analogs in liquid and solid powder drugs.

Key words: liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS), high resolution mass spectrometry (HRMS), fentanyl and its analogues, drugs, screening

邓慧芬, 张建莹, 卞学海, 罗耀, 赵祥龙. 液相色谱-四极杆/飞行时间质谱法快速筛查测定药物中芬太尼类物质[J]. 色谱, 2020, 38(8): 906-913.

DENG Huifen, ZHANG Jianying, BIAN Xuehai, LUO Yao, ZHAO Xianglong. Rapid screening and determination of fentanyl and its analogues in drugs by liquid chromatography- quadrupole time-of-flight mass spectrometry[J]. Chinese Journal of Chromatography, 2020, 38(8): 906-913.

图/表 7

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No.	Compound	Chemical formula	Parent ions		Mass accuracy/10^-6 (ppm)	Fragment 1	Fragment 2	t_R/min
No.	Compound	Chemical formula	Theoretical mass	Experimental mass	Mass accuracy/10^-6 (ppm)	Fragment 1	Fragment 2	t_R/min
1	N-phenethyl-4-piperidone	C₁₃H₁₇NO	204.138	204.1381	-0.9	77.222	103.169	1.71
2	despropionylfentanyl (4-ANPP)	C₁₉H₂₄N₂	281.201	281.2009	-1	105.169	188.111	9.06
3	acetyl fentanyl	C₂₁H₂₆N₂O	323.212	323.2116	-0.6	105.183	132.183	8.13
4	acrylfentanyl	C₂₂H₂₆N₂O	335.212	335.2114	-1.2	188.097	105.151	10.34
5	fentanyl	C₂₂H₂₈N₂O	337.227	337.2271	-1	105.169	188.111	10.93
6	acetyl-α-methylfentanyl	C₂₂H₂₈N₂O	337.227	337.2272	-0.8	259.169	208.183	9.27
7	thiofentanyl	C₂₀H₂₆N₂OS	343.184	343.1835	-1	194.111	111.111	9.64
8	cis-3-methylfentanyl	C₂₃H₃₀N₂O	351.243	351.2429	-0.7	105.169	202.111	12.90
9	isobutyryl fentanyl	C₂₃H₃₀N₂O	351.243	351.2427	-1	105.151	188.111	13.19
10	α-methyl fentanyl	C₂₃H₃₀N₂O	351.243	351.2428	-0.9	202.169	91.222	11.98
11	β-hydroxy fentanyl	C₂₂H₂₈N₂O₂	353.222	353.2219	-1.4	192.040	285.111	8.91
12	p-fluorofentanyl	C₂₂H₂₇FN₂O	355.218	355.2178	-0.7	105.169	188.111	11.50
13	3-methylthiofentanyl	C₂₁H₂₈N₂OS	357.200	357.1994	-0.4	111.111	160.111	11.66
14	α-methylthiofentanyl	C₂₁H₂₈N₂OS	357.200	357.1991	-1.1	91.151	119.097	10.70
15	β-hydroxythiofentanyl	C₂₀H₂₆N₂O₂S	359.179	359.1787	-0.3	186.111	204.111	7.75
16	valeryl fentanyl	C₂₄H₃₂N₂O	365.259	365.2580	-2.1	105.151	188.111	15.36
17	ohmefentanyl	C₂₃H₃₀N₂O₂	367.238	367.2377	-0.7	200.111	218.040	10.54
18	butyryl fentanyl	C₂₃H₂₉FN₂O	369.234	369.2333	-0.9	105.183	188.111	13.77
19	4-fluoro-isobutyryl fentanyl	C₂₃H₂₉FN₂O	369.234	369.2336	-0.3	105.169	188.111	14.07
20	p-fluorobutyryl fentanyl	C₂₃H₂₉FN₂O	369.234	369.2336	-0.3	105.151	188.111	14.07
21	ocfentanil	C₂₂H₂₇FN₂O₂	371.213	371.2126	-0.8	105.183	188.111	8.21
22	furanyl fentanyl	C₂₄H₂₆N₂O₂	375.207	375.2064	-0.9	105.183	188.111	11.76
23	remifentanil	C₂₀H₂₈N₂O₅	377.207	377.2068	-0.8	285.111	317.111	6.44
24	tetrahydrofuran fentanyl	C₂₄H₃₀N₂O₂	379.238	379.2376	-1	188.111	105.165	9.60
25	sufentanil	C₂₂H₃₀N₂O₂S	387.210	387.2095	-1.4	111.111	238.040	14.55
26	carfentanyl	C₂₄H₃₀N₂O₃	395.233	395.2327	-0.6	246.111	335.151	12.83
27	alfentanil	C₂₁H₃₂N₆O₃	417.261	417.2605	-0.9	197.111	268.111	10.85

No.	Compound	Chemical formula	Parent ions		Mass accuracy/10^-6 (ppm)	Fragment 1	Fragment 2	t_R/min
No.	Compound	Chemical formula	Theoretical mass	Experimental mass	Mass accuracy/10^-6 (ppm)	Fragment 1	Fragment 2	t_R/min
1	N-phenethyl-4-piperidone	C₁₃H₁₇NO	204.138	204.1381	-0.9	77.222	103.169	1.71
2	despropionylfentanyl (4-ANPP)	C₁₉H₂₄N₂	281.201	281.2009	-1	105.169	188.111	9.06
3	acetyl fentanyl	C₂₁H₂₆N₂O	323.212	323.2116	-0.6	105.183	132.183	8.13
4	acrylfentanyl	C₂₂H₂₆N₂O	335.212	335.2114	-1.2	188.097	105.151	10.34
5	fentanyl	C₂₂H₂₈N₂O	337.227	337.2271	-1	105.169	188.111	10.93
6	acetyl-α-methylfentanyl	C₂₂H₂₈N₂O	337.227	337.2272	-0.8	259.169	208.183	9.27
7	thiofentanyl	C₂₀H₂₆N₂OS	343.184	343.1835	-1	194.111	111.111	9.64
8	cis-3-methylfentanyl	C₂₃H₃₀N₂O	351.243	351.2429	-0.7	105.169	202.111	12.90
9	isobutyryl fentanyl	C₂₃H₃₀N₂O	351.243	351.2427	-1	105.151	188.111	13.19
10	α-methyl fentanyl	C₂₃H₃₀N₂O	351.243	351.2428	-0.9	202.169	91.222	11.98
11	β-hydroxy fentanyl	C₂₂H₂₈N₂O₂	353.222	353.2219	-1.4	192.040	285.111	8.91
12	p-fluorofentanyl	C₂₂H₂₇FN₂O	355.218	355.2178	-0.7	105.169	188.111	11.50
13	3-methylthiofentanyl	C₂₁H₂₈N₂OS	357.200	357.1994	-0.4	111.111	160.111	11.66
14	α-methylthiofentanyl	C₂₁H₂₈N₂OS	357.200	357.1991	-1.1	91.151	119.097	10.70
15	β-hydroxythiofentanyl	C₂₀H₂₆N₂O₂S	359.179	359.1787	-0.3	186.111	204.111	7.75
16	valeryl fentanyl	C₂₄H₃₂N₂O	365.259	365.2580	-2.1	105.151	188.111	15.36
17	ohmefentanyl	C₂₃H₃₀N₂O₂	367.238	367.2377	-0.7	200.111	218.040	10.54
18	butyryl fentanyl	C₂₃H₂₉FN₂O	369.234	369.2333	-0.9	105.183	188.111	13.77
19	4-fluoro-isobutyryl fentanyl	C₂₃H₂₉FN₂O	369.234	369.2336	-0.3	105.169	188.111	14.07
20	p-fluorobutyryl fentanyl	C₂₃H₂₉FN₂O	369.234	369.2336	-0.3	105.151	188.111	14.07
21	ocfentanil	C₂₂H₂₇FN₂O₂	371.213	371.2126	-0.8	105.183	188.111	8.21
22	furanyl fentanyl	C₂₄H₂₆N₂O₂	375.207	375.2064	-0.9	105.183	188.111	11.76
23	remifentanil	C₂₀H₂₈N₂O₅	377.207	377.2068	-0.8	285.111	317.111	6.44
24	tetrahydrofuran fentanyl	C₂₄H₃₀N₂O₂	379.238	379.2376	-1	188.111	105.165	9.60
25	sufentanil	C₂₂H₃₀N₂O₂S	387.210	387.2095	-1.4	111.111	238.040	14.55
26	carfentanyl	C₂₄H₃₀N₂O₃	395.233	395.2327	-0.6	246.111	335.151	12.83
27	alfentanil	C₂₁H₃₂N₆O₃	417.261	417.2605	-0.9	197.111	268.111	10.85

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