基于薄层色谱原位富集显微拉曼光谱法快速检测降压保健食品中非法添加的3种西药

doi:10.7503/cjcu20170735

摘要/Abstract

摘要：

基于薄层色谱(TLC)与显微拉曼光谱(Micro-Raman spectroscopy)联用技术, 对降压类保健食品中非法添加的3种化学成分(尼莫地平、拉西地平和尼群地平)进行原位富集检测, 并对6种降压保健食品进行了测试. TLC法采用硅胶60F₂₅₄铝合金薄层板为固定相, 环己烷-乙酸乙酯(体积比5∶4)混合溶液为展开剂, 在紫外灯(254 nm)下检视定位, 使添加成分与保健食品基质进行初步分离; 再对薄层板上微量添加成分斑点用丙酮原位富集, 直接检测富集斑点的显微拉曼光谱, 激光光源波长780 nm, 显微镜倍数10倍, 扫描范围100~3300 cm^-1, 激光功率24.0 mW, 曝光时间0.05000 s, 扫描30次. 结果表明, 常用降压类保健食品基质对添加成分检测结果无干扰, 检出限为2~4 μg. 本方法准确、灵敏、简便、快速, 为降压类保健食品中非法添加化学成分的快速检测提供了新的手段.

关键词: 薄层色谱, 原位富集, 显微拉曼光谱, 降压类保健食品, 非法添加

Abstract:

A thin layer chromatography(TLC) combined with micro-Raman spectroscopy technology was applied for the detection of three chemical components(nimodipine, lacidipine and nitrendipine) added illegally into the antihypertensive health food by concentration in situ and six kinds of antihypertensive health food products were tested. The doping ingredient and substrate were separated initially by TLC with silica gel 60F₂₅₄ aluminum alloy plates as the stationary phase, [V(cyclohexane)∶V(acetic ether)=5∶4] as the developing solvent, and using UV light(254 nm) to view and locate. Then the spots of the trace of doping ingredient were concentrated on the TLC plate with acetone, and the concentrated points were detected directly by the micro-Raman spectroscopy with laser source of 780 nm, objective of 10×, spectral range of 100—3300 cm^-1, laser power of 24.0 mW, exposure time of 0.05000 s and number of scans of 30. The results show the common substrate in the antihypertensive health food products had no interference with the detecting results of doping components. The LOD was 2—4 μg. The method is accurate, sensitive, simple and rapid, and can provide a new reference method for the technology research of rapid detection of the ingredients added illegally in the antihypertensive health food.

Key words: Thin layer chromatography(TLC), Concentration in situ, Micro-Raman spectroscopy, Antihypertensive health food, Added illegally

中图分类号:

O657.37

TrendMD:

许凤, 李晓雪, 李莉, 刘吉成. 基于薄层色谱原位富集显微拉曼光谱法快速检测降压保健食品中非法添加的3种西药. 高等学校化学学报, 2018, 39(6): 1172.

XU Feng, LI Xiaoxue, LI Li, LIU Jicheng. Rapid Detection of Three Chemical Components Added Illegally into Antihypertensive Health Food by TLC Concentration in situ Combined with Micro-Raman Spectroscopy Method^†. Chem. J. Chinese Universities, 2018, 39(6): 1172.

图/表 8

Fig.1 Raman spectra of powder and TLC in situ of reference substances A, B and C(A—C)a. Powder; b. TLC in situ; c. developing solvent on silica gel 60F254 aluminum alloy plates; d. acetone on silica gel 60F254 aluminum alloy plates.

Table 1 Characteristic peaks and belonging in Raman spectrum of reference substance

Analyte	Structure	Frequency/cm^-1	Peak strength	Vibrational mode
Nimodipine(A)		1641.30	s	$ν CC$
		1345.69	s	Deformation vibration of CH
		1269.85	m	ν_ring
		1138.41	m	ν_C—C
		827.50	m	ν_skeleton
		Lacidipine(B)		1676.82	s	$ν CC$
1451.85	s			Deformation vibration of CH₂ and CH₃
1312.83	m			$β C H 2$
Nitrendipine(C)				1646.49	s	$ν CC$
		1578.24,1487.24	m	ν_{pyriding ring}
		1348.21	vs	β_CH
		1262.27	w	$ν s (N O 2)$
		1214.24	m	ν_C—O—C
		1004.44	s	Respiratory vibration of the triangle of benzene ring
		824.97	m	ν_skeleton
		807.28	m	γ_CH(1,3 disubstituted benzene ring )
		592.42	m	β_{pyriding ring}

Table 1 Characteristic peaks and belonging in Raman spectrum of reference substance

Analyte	Structure	Frequency/cm^-1	Peak strength	Vibrational mode
Nimodipine(A)		1641.30	s	$ν CC$
		1345.69	s	Deformation vibration of CH
		1269.85	m	ν_ring
		1138.41	m	ν_C—C
		827.50	m	ν_skeleton
		Lacidipine(B)		1676.82	s	$ν CC$
1451.85	s			Deformation vibration of CH₂ and CH₃
1312.83	m			$β C H 2$
Nitrendipine(C)				1646.49	s	$ν CC$
		1578.24,1487.24	m	ν_{pyriding ring}
		1348.21	vs	β_CH
		1262.27	w	$ν s (N O 2)$
		1214.24	m	ν_C—O—C
		1004.44	s	Respiratory vibration of the triangle of benzene ring
		824.97	m	ν_skeleton
		807.28	m	γ_CH(1,3 disubstituted benzene ring )
		592.42	m	β_{pyriding ring}

Table 2 Main ingredients of 6 kinds of antihypertensive health food

No.	Lot number	Main ingredient	Dose(grain/time)
1	421297004	Fish oil, gelatin, natural vitamin E, glycerin, purified water	1 grain
2	427307008	Soybean lecithin, soybean oil, gelatin, purified water, glycerin, caramel pigment	1 grain
3	BBJ60200101	Fish oil, gelatin, natural vitamin E, glycerin, water	1 grain
4	11240039	Lobute, lobute, green tea	1 bag
5	20170103	Ginkgo fruit, pingju chrysanthemum, pueraria, flavin, chicory, corn whiskers	1 bag
6	20151101	Hawthorn	3 g

Fig.2 TLC result of A, B, C(A) and Raman spectra of TLC in situ(B)RA, RB, RC: A, B, C reference substance; RA,B,C: mixture of A, B, C reference substance; +: simulated positive sample; -: negative sample.

Fig.3 TLC result(A) and Raman spectra of different deposition amount(μg) of reference substances A, B and C(B—D)

Table 3 Possible addition amount of three kinds of chemical composition in antihypertensive health food

Doping component	Pharmaceutical dosage form	Preparations minimum specification	Possible amount of doping	Sample solution	Reference solution	TLC spotting volume(sample or reference)	TLC spotting deposited amount^[20]	LOD
Nimodipine	Nimodipine tablet	20 mg/Tablet	40 mg	A dose/5 mL	40 mg/5 mL	6 μL	48 μg	4 μg
Lacidipine	Lacidipine tablet	4 mg/Tablet	4 mg	A dose/5 mL	4 mg/5 mL	6 μL	4.8 μg	2 μg
Nitrendipine	Nitrendipine tablet	10 mg/Tablet	10 mg	A dose/5 mL	10 mg/5 mL	6 μL	12 μg	4 μg

Fig.4 TLC results of 6 kinds of antihypertensive health foodRA, RB, RC: A, B, C reference substance; S1—S6: samples 1—6 of antihypertensive health food.

Fig.5 Raman spectra of samples 2, 3 and 5 in the same Rf as A reference substance(A) and B reference substance(B)

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