Ovoglycoprotein-Bonded HPLC Stationary Phases for Chiral Recognition

Abstract

Commercial chicken ovomucoid (OMCHI) and OMCHI, isolated by precipitation of egg whites with organic solvents, both of which as crude products, were fractionated by anion- and cation-exchange chromatography. The obtained four fractions were characterized by reversed-phase chromatography, N-terminal sequencing, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry, determination of sugar contents, and trypsin-inhibitory activities. Three fractions were OMCHI variants differing in carbohydrate composition, especially in sialic acid content, and the other fraction was tentatively termed ovoglycoprotein (OGCHI). The OMCHI and OGCHI are different in physicochemical and biochemical properties: average molecular weight 26000-27700 for OMCHI variants and 29700 for OGCHI; N-terminal amino acid, Ala for OMCHI and Thr for OGCHI; and trypsin-inhibitory activity, positive for OMCHI and negative for OGCHI. These OMCHI variants and OGCHI were bound to aminopropyl silica gels to evaluate chiral recognition ability. OMCHI is reported to have chiral recognition ability (Miwa, T.; et al. Chem. Pharm. Bull. 1987, 35, 682-686). However, neither OMCHI variant had appreciable chiral recognition ability, while the OGCHI had excellent chiral recognition properties as compared to those of the OMCHI reported previously. This reveals that the chiral recognition ability of the OMCHI reported previously comes from the OGCHI, which is present in crude OMCHI as an impurity.

Chiral separation by chromatographic and electromigration techniques. A Review

“…Phases of this type showed enantioselectivity for benzodiazepines and 2-arylpropionic acid derivatives. Haginaka et al [209] isolated a new ovoglycoprotein, OGCHI from chicken egg white and bonded it to aminopropyl-silica gel [210]. While this CSP showed enantioselectivity preferably for basic compounds, a newly developed CSP prepared from Japanese quail egg white was found to be more suitable for acidic compounds [211].…”

Resolution of Dihydropyridine Calcium Antagonist Enantiomers Using HPLC with Ovoglycoprotein as a Chiral Stationary Phase

“…12 To get OGCHI, the obtained OMCHI was further purified on SP sepharose FF and Sephadex G-25 (fine) columns as reported previously. 11 A reversed-phase chromatography 11 was employed to determine the purity of the isolated OGCHI and the amounts of OGCHI bound to activated silica gels. The purity of OGCHI was estimated to be 99%, calculated from peak area.…”

“…11 The obtained fractions were composed of three OMCHI variants differing in carbohydrate composition and a new component, ovoglycoprotein (ovoglycoprotein from chicken egg whites, OGCHI). It is found that none of the OMCHI variants give any appreciable chiral recognition ability and that the chiral recognition ability of crude OMCHI comes from OGCHI, which has an average molecular weight of 29700.…”

HPLC Chiral Stationary Phases Produced with Isolated Human Serum Albumin Fragments

“…This property has led to the development of a number of HPLC chiral stationary phases based upon albumins such as bovine serum albumin (BSA) 1 and human serum albumin (HSA), 2 glycoproteins such as α1-acid glycoprotein, 3,4 ovomucoid, 5 ovoglycoprotein, 6 avidin 7 and cellulase 8 and enzymes such as trypsin, 9 α-chymotrypsin, 10 lysozyme 11 and pepsin. 12 Disadvantages of protein-based HPLC columns have included low capacity, lack of column ruggedness in some instances and a limited understanding of the chiral recognition mechanisms.…”