磺丁基醚-β-环糊精修饰毛细管电色谱拆分地平类药物对映体

doi:10.7503/cjcu20140099

摘要/Abstract

摘要：

采用甲基丙烯酸缩水甘油酯原位聚合物基质, 将磺丁基醚-β-环糊精修饰到毛细管内壁, 制得了一种毛细管电色谱手性柱(SECDP), 并通过红外光谱(IR)和扫描电子显微镜(SEM)表征了其结构. 磺酸基可提供足够稳定的正向电渗流(EOF), 基于磺丁基醚-β-环糊精在固定相和流动相中的协同作用, 通过优化手性添加剂浓度、 pH值、施加电压、温度及有机调节剂含量等条件, 利用该开管电色谱柱拆分了氨氯地平、尼莫地平和尼卡地平等10种地平类手性药物对映体. 优化的流动相组成为20 mmol/L NaH₂PO₄(pH=4.0), 含4.0 mmol/L 磺丁基醚-β-环糊精, 乙腈的体积分数为10%~25%, 施加电压15~25 kV, 温度为15 ℃, 电动进样2 kV×5 s, 检测波长为236 nm. 在上述条件下, 分离度(R_S)可达3.62, 柱效达61011块/m, 分析时间一般为6~15 min. 基于色谱分离数据, 探讨了相关的手性分离机理.

关键词: 毛细管电色谱法, 甲基丙烯酸缩水甘油酯原位聚合物基质, 磺丁基醚-β, -环糊精, 手性分离, 地平类药物

Abstract:

A novel sulfobutyl ether-β-cyclodextrin(SBE-β-CD) capillary electrochromatographic column(SECDP) was prepared by glycidyl methacrylate polymer matrix in situ and its modification. Its basic structure was characterized by infrared spectroscopy(IR) and scanning electron microscopy(SEM). The sulfonic acid group could provide a enough and stable positive electroosmotic flow(EOF). Enantioseparations of 10 kinds of chiral dihydropyridine drugs including amlodipine, nimodipine, nicardipine and so on were achieved by synergistic effects of SBE-β-CD in the stationary and mobile phases. Some effect factors such as the chiral additive concentration, pH value, the applied voltage, temperature and organic modifier content were optimized. The optimal separation conditions were as follows: the mobile phase consisting of 20 mmol/L NaH₂PO₄(pH=4.0), containing 4.0 mmol/L of SBE-β-CD, acetonitrile volume content of 10% to 25%, an applied voltage of 15—25 kV, the temperature at 15 ℃, electrokinetic injection 2 kV×5 s and the detection wavelength at 236 nm. Under the above conditions, the separation resolutions(R_s) of dihydropyridine enantiomers were up to 3.62, the column efficiency were about 61011/m, and the analysis time were generally between 6—15 min. Based on the data of chromatographic separations, the related chiral separation mechanism was also discussed. The preparation method of SECDP and chiral separation method were rapid, simple, reprodu-cible and suitable for fast enantioseparations and montoring of chiral dihydropyridine drugs.

Key words: Capillary electrochromatography, Glycidyl methacrylate polymer matrix in situ, Sulfobutyl ether-β, -cyclodextrin, Chiral separation, Dihydropyridine drug

中图分类号:

O657

TrendMD:

聂桂珍, 李来生, 程彪平, 周仁丹, 张宏福. 磺丁基醚-β-环糊精修饰毛细管电色谱拆分地平类药物对映体. 高等学校化学学报, 2014, 35(7): 1414.

NIE Guizhen, LI Laisheng, CHENG Biaoping, ZHOU Rendan, ZHANG Hongfu. Enantioseparations of Dihydropyridine Drugs by Sulfobutyl Ether-β-Cyclodextrin Modified Capillary Electrochromatography^†. Chem. J. Chinese Universities, 2014, 35(7): 1414.

图/表 10

Fig.1 Chemical structures of dihydropyridine drugs

Fig.2 Preparation route of open-tublar capillary column modified with sulfbutyl ether-β-cyclodextrin

Fig.3 FTIR spectra of SECDP(a) and GMA-EDMA(b) before modification

Fig.4 SEM images of inner wall in capillary column before(A) and after(B) modification

Fig.5 Effects of pH value(A) and organic solvent content(B) on the EOF

Table 1 Effects of pH values on the separationparameters

pH	t₁	t₂	α	R_S
2.5	9.03	10.14	1.46	1.85
3.5	7.34	7.88	1.45	1.84
4.0	6.38	7.55	1.46	1.84
4.5	4.22	4.63	1.18	1.24
6.5	3.90	3.90	1.00	0

Fig.6 Effect of acetonitrile content on the enantioseparation of amlodipine besylate a. 18%; b. 22%; c. 25%. d. 30%. Conditions: 20 mmol/L NaH2PO4(pH=4.0), 4.0 mmol/L SBE-β-CD, applied voltage 20 kV and temperature 15 ℃, injection: 2 kV×5 s.

Table 2 Reproducibilities of amlodipine enantiomers about retention time and peak area(n=5)

	RSD(%)
	t₁	t₂	A₁	A₂
Intra-day	1.09	1.24	4.35	4.48
Inter-day	1.21	1.56	5.54	5.65

Table 3 Enantioseparation results of dihydropyridine drugs*

Compound	t₁/min	t₂/min	R_s	N₁/(Plates·m^-1)	N₂/(Plates·m^-1)	ACN(%)	V/kV
Amlodipine besylate	6.38	7.55	1.84	22976	20054	22	20
Amlodipine mesylate	4.64	4.93	1.75	30261	29851	22	25
Nimodipine	7.04	7.87	3.62	61011	60034	20	25
Nicardipine	5.33	6.03	3.52	50142	49867	15	25
Manidipine	10.97	11.75	1.52	37072	35721	25	15
Benidipine	6.23	6.64	1.54	54853	52014	10	20
Cilnidipine	7.37	8.16	1.50	13795	12876	25	25
Nitredipine	11.88	12.56	1.15	25603	20230	10	15
Felodipine	15.90	16.76	1.14	12065	10274	18	25
Lercanidipine	5.70	6.13	0.63	11254	10235	12	25
Lacidipine	29.12	29.83	0.32	10242	10112	12	20

Fig.7 Electrochromatograms of some dihydropyridine enantiomers on SECDP (A) Amlodipine besytle; (B) amlodipine mesylate; (C) nimodipine; (D) nicardipine; (E) manidipine; (F) benidipine; (G) cilnidipine.

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