Molecularly Imprinted Hollow Spheres for the Solid Phase Extraction of Protein†

doi:10.7503/cjcu20180327

Abstract

Abstract:

Molecularly imprinted polymer was widely used as the stuffing of solid phase extraction(SPE) due to its high selectivity. Hb isolation from complex matrix was established based on molecularly imprinted solid phase extraction(MISPE). Mono-dispersed molecularly imprinted hollow spheres(MIHS) of hemoglobin(Hb) were synthesized using silica nanospheres as the sacriﬁcial matrix. It is proved that the MIHS with good monodispersion and uniform particle size by TEM scanning was suitable for SPE enrichment matrix. Therefore, the extraction of Hb, bovine serum albumin(BSA) and cytochrome C(Cyt C) by MIHSs was investigated by SPE combined with liquid chromatographic method. The recovery of Hb was higher than 80% in phosphate buffer solution(pH=7) while it was lower than 50% for RNase A and Cyt C. Meanwhile, the degree of precision(RSD) was lower than 5%. Compared to the surface imprinted silica(SIS), the MIHS had good recognition and fast adsorption-desorption dynamics for Hb. Moreover, this method was also confirmed in real sample, and the adsorption of Hb was 82% in the spiked bovine serum.

Key words: Solid phase extraction, Hemoglobin(Hb), Molecular imprinting, Hollow sphere

CLC Number:

O657

TrendMD:

MENG Zihui,WANG Yifei,XIE Tengsheng,CHEN Wei,QIU Lili,XUE Min. Molecularly Imprinted Hollow Spheres for the Solid Phase Extraction of Protein^†[J]. Chem. J. Chinese Universities, 2019, 40(1): 62.

Figures/Tables 10

Scheme 1 Schematic diagram for the fabrication of SIS

Scheme 2 Schematic diagram for the fabrication of MIHS

Fig.1 TEM images of silica particle(A), SIS(B)and MIHS(C, D)

Fig.2 Adsorption of Hb on MIHS(a), NIHS(b), SIS(c) and NIS(d)

Fig.3 Recoveries of Hb on MIHS columns using different eluatesa. Formic acid/ethanol solution; b. 10% SDS solution; c. 10% TritonX-100 solution; d. 10% Tris solution.

Fig.4 Recoveries of Hb with different flow rate

Table 1 Recoveries of protein by MIHS-SPE and SIS-SPE

Protein type	Sample concentration/ (mg·mL^-1)	SIS		MIHS
Protein type	Sample concentration/ (mg·mL^-1)	Recovery(%)	RSD(%)	Recovery(%)	RSD(%)
Hb	5	85.4	4.75	82.3	4.56
RNase A	5	43.2	3.82	47.3	4.61
Cyt C	5	37.1	4.87	40.5	3.84
Hb	10	82.6	4.68	81.5	5.64
RNase A	10	25.6	3.32	32.8	3.15
Cyt C	10	34.2	2.37	26.4	4.12
Hb	25	60.4	4.68	61.3	4.02
RNase A	25	25.3	4.81	33.1	4.03
Cyt C	25	32.5	4.64	37.4	3.05

Fig.5 Liquid chromatograms of Hb, RNase A and Cyt C before and after SPE

Fig.6 Liquid chromatograms of bovine serum(A), bovine serum spiked with Hb(B) and extracted solution(C)

Table 2 Recoveries of Hb real sample by MIHS-SPE

Sample concentration/(mg·mL^-1)	Recovery(%)	RSD(%)
5	82.9	4.23
10	82.3	5.45
25	60.3	3.91

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