二脲基桥联β-环糊精手性液相色谱键合相的制备与性能评价

doi:10.3724/SP.J.1123.2019.08006

摘要/Abstract

摘要：

采用六亚甲基二异氰酸酯与6-脱氧-6-羟乙基胺基-β-环糊精反应,合成二脲基桥联β-环糊精,并将其键合到硅胶表面,制备一种新型的二脲基桥联β-环糊精固定相(UBCDP)。通过红外光谱、质谱、元素分析等进行结构表征,以黄烷酮类、氨基酸类、三唑类手性药物和农药为探针,评价其手性色谱性能,探讨相关分离机理。进行色谱条件优化,并与单β-环糊精固定相(CDCSP)比较。试验了多种手性化合物,其中25个被拆分,2'-羟基黄烷酮、己唑醇、丹磺酰亮氨酸等分离度(R_s)达到1.52~4.35,且能拆分体积较大的橙皮甙。这与桥联β-环糊精的协同包结作用有关。而CDCSP仅能拆分少量的对映体,且分离度较低。UBCDP手性拆分能力更强,在手性药物和农药监测中有应用价值。

关键词: 高效液相色谱法, 二脲基桥联β-环糊精固定相, 手性药物与农药拆分, 色谱性能评价, 色谱分离机理

Abstract:

A diureido bridged β-cyclodextrin dimer was synthesized by the reaction of 6-deoxy-6-hydroxyethylamino-β-cyclodextrin with hexamethylene diisocyanate. The dimer was then bonded onto silica to obtain a novel diureido bridged β-cyclodextrin-bonded stationary phase (UBCDP). The structure of diureido bridged stationary phase was characterized by infrared spectroscopy, mass spectrometry, and elemental analysis. By using racemic drugs and pesticides as probes (flavanones, triazoles, and dansylated amino acids), the chromatographic properties of UBCDP were evaluated. The separation mechanism was elucidated by comparison with a single β-cyclodextrin chiral stationary phase (CDCSP). The composition, pH value of the mobile phase and column temperature were optimized. The results showed that the UBCDP can resolve 25 chiral compounds, especially 2'-hydroxyflavanone, hexaconazole and dansyl leucine with high resolutions (R_s, 1.52-4.35), and the larger volume hesperidin could also be separated. This trend was related to the synergistic inclusion of bridged cyclodextrins. CDCSP can separate only a small number of enantiomers with low resolutions. UBCDP exhibited better enantio-separation ability and hence had potential application in chiral drugs and chiral pesticide residues analysis.

Key words: high performance liquid chromatography (HPLC), diureido bridged β-cyclodextrin-bonded stationary phase (UBCDP), chiral drug and pesticide separations, chromatographic performance evaluation, chromatographic separation mechanism

中图分类号:

O658

双亚洲, 王惠, 张天赐, 李来生. 二脲基桥联β-环糊精手性液相色谱键合相的制备与性能评价[J]. 色谱, 2020, 38(4): 464-475.

SHUANG Yazhou, WANG Hui, ZHANG Tianci, LI Laisheng. Preparation and evaluation of diureido bridged β-cyclodextrin-bonded chiral stationary phase by high-performance liquid chromatography[J]. Chinese Journal of Chromatography, 2020, 38(4): 464-475.

图/表 14

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Compound	k₁	k₂	k₃	R_s1	R_s2	Elution order	CSP
Nitroaniline	1.80	1.96	5.13	1.45	17.78	m-o-p	UBCDP
	0.80	1.27	3.84	2.40	11.44	m-o-p	CDCSP
Phenylenediamine	0.16	0.66	1.41	4.58	3.68	m-p-o	UBCDP
	0.20	/	/	/	/	/	CDCSP
Phthalic acid	0.69	2.67	8.11	17.15	23.68	m-p-o	UBCDP
	1.70	2.37	7.42	8.93	10.57	m-p-o	CDCSP

Compound	k₁	k₂	k₃	R_s1	R_s2	Elution order	CSP
Nitroaniline	1.80	1.96	5.13	1.45	17.78	m-o-p	UBCDP
	0.80	1.27	3.84	2.40	11.44	m-o-p	CDCSP
Phenylenediamine	0.16	0.66	1.41	4.58	3.68	m-p-o	UBCDP
	0.20	/	/	/	/	/	CDCSP
Phthalic acid	0.69	2.67	8.11	17.15	23.68	m-p-o	UBCDP
	1.70	2.37	7.42	8.93	10.57	m-p-o	CDCSP

Compound	k₁	k₂	α	R_s	Mobile phase	Column
Flavanone	3.57	4.02	1.12	1.47	1%HCOOH/ACN (90/10, v/v)	UBCDP
	3.13	/	/	0	1%HCOOH/ACN (80/20, v/v)	CDCSP
2'-Hydroxy flavanone	2.33	3.27	1.40	4.35	1%HCOOH/MeOH (75/25, v/v)	UBCDP
	3.34	3.53	1.05	1.67	1%HCOOH/MeOH (85/15, v/v)	CDCSP
4'-Hydroxy flavanone	3.61	4.38	1.22	3.47	1%HCOOH/ACN (90/10, v/v)	UBCDP
	9.59	10.01	1.04	0.66	1%HCOOH/MeOH (85/15, v/v)	UBCDP
	4.88	/	/	0	1%HCOOH/ACN (80/20, v/v)	CDCSP
6-Hydroxy flavanone	5.18	5.59	1.08	1.14	1%HCOOH/MeOH (85/15, v/v)	UBCDP
	9.89	/	/	0	1%HCOOH/MeOH (90/10, v/v)	CDCSP
6-Methoxy flavanone	6.88	7.33	1.07	1.06	1%HCOOH/MeOH (70/30, v/v)	UBCDP
	6.57	/	/	0	1%HCOOH/MeOH (90/10, v/v)	CDCSP
Naringin	2.61	3.18	1.22	2.73	1%HCOOH/MeOH (80/20, v/v)	UBCDP
	4.12	4.54	1.10	1.12	1%HCOOH/MeOH (80/20, v/v)	CDCSP
Hesperidin	2.45	2.81	1.15	1.34	1%HCOOH/ACN (90/10, v/v)	UBCDP
	4.83	/	/	0	1%HCOOH/MeOH(80/20, v/v)	CDCSP
Catechin	3.29	3.82	1.16	2.46	1%HCOOH/ACN (90/10, v/v)	UBCDP
	4.16	4.62	1.19	1.36	1%HCOOH/ACN (85/15, v/v)	CDCSP

Compound	k₁	k₂	α	R_s	Mobile phase	Column
Flavanone	3.57	4.02	1.12	1.47	1%HCOOH/ACN (90/10, v/v)	UBCDP
	3.13	/	/	0	1%HCOOH/ACN (80/20, v/v)	CDCSP
2'-Hydroxy flavanone	2.33	3.27	1.40	4.35	1%HCOOH/MeOH (75/25, v/v)	UBCDP
	3.34	3.53	1.05	1.67	1%HCOOH/MeOH (85/15, v/v)	CDCSP
4'-Hydroxy flavanone	3.61	4.38	1.22	3.47	1%HCOOH/ACN (90/10, v/v)	UBCDP
	9.59	10.01	1.04	0.66	1%HCOOH/MeOH (85/15, v/v)	UBCDP
	4.88	/	/	0	1%HCOOH/ACN (80/20, v/v)	CDCSP
6-Hydroxy flavanone	5.18	5.59	1.08	1.14	1%HCOOH/MeOH (85/15, v/v)	UBCDP
	9.89	/	/	0	1%HCOOH/MeOH (90/10, v/v)	CDCSP
6-Methoxy flavanone	6.88	7.33	1.07	1.06	1%HCOOH/MeOH (70/30, v/v)	UBCDP
	6.57	/	/	0	1%HCOOH/MeOH (90/10, v/v)	CDCSP
Naringin	2.61	3.18	1.22	2.73	1%HCOOH/MeOH (80/20, v/v)	UBCDP
	4.12	4.54	1.10	1.12	1%HCOOH/MeOH (80/20, v/v)	CDCSP
Hesperidin	2.45	2.81	1.15	1.34	1%HCOOH/ACN (90/10, v/v)	UBCDP
	4.83	/	/	0	1%HCOOH/MeOH(80/20, v/v)	CDCSP
Catechin	3.29	3.82	1.16	2.46	1%HCOOH/ACN (90/10, v/v)	UBCDP
	4.16	4.62	1.19	1.36	1%HCOOH/ACN (85/15, v/v)	CDCSP

Compound	k₁	k₂	α	R_s	Mobile phase	Column
Dns-Leu	1.52	1.79	1.17	1.69	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.16	1.21	1.04	0.79	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Phe	5.13	6.42	1.25	2.07	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.75	1.82	1.04	0.87	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Asp	2.63	2.97	1.12	1.32	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.24	1.34	1.08	0.82	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Met	1.91	2.14	1.12	1.25	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.49	1.58	1.06	0.70	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Tyr	1.11	1.18	1.07	0.78	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.37	/	/	/	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Thr	2.08	2.37	1.62	0.73	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.37	/	/	<0.50	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Ser	4.01	4.07	1.01	0.63	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	1.47	/	/	<0.50	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP
Dns-Ala	3.34	3.37	1.01	0.85	1% TEAA/MeOH/ACN (70/25/5, v/v/v, pH 4.0)	UBCDP
	2.19	2.21	1.01	0.65	1% TEAA/MeOH/ACN (80/15/5, v/v/v, pH 4.0)	CDCSP