Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
38, Number 12, December 2000, pp. 517-520
Study
on Open Tubular Capillary Affinity Liquid Chromatography
Mingliang
Ye, Hanfa Zou, Zhen Liu, Zhengdeng Lei, and Jianyi Ni
National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics,
Chinese Academy of Sciences, Dalian 116011, China
A
novel mode of affinity chromatography (AC) based on an open tubular capillary
column (OTAC) is demonstrated. The OTAC column is prepared by immobilizing Cibacron
blue F3GA onto the inner surface of a 50-µm-i.d. capillary column. The
AC experiment is performed on a capillary electrophoresis instrument by using
its pressure system as the driving force. Bovine serum albumin and lysozyme
(Lys) are successfully separated with stepwise gradient elution. The relative
standard deviation (RSD) for the elution time of the retained Lys is 0.08%,
and good repeatability of its peak area and peak height with an RSD value lower
than 2.12% for 10 consecutive runs is observed. The loading capacity and detection
limit for the retained Lys are approximately 36 ng and 8.6 ng, respectively.
It is also found that the amount of protein adsorbed is unaffected by the flow
rate of the loading buffer, and OTAC can be used for the fast determination
of biopolymers. Some of the advantages of OTAC over conventional modes of open
tubular capillary liquid chromatography are that the detection sensitivity and
loading capacity of a sample can be greatly improved, because the relatively
large inner diameter of the capillary can be adopted and the whole capillary
column can be used to adsorb the solute in OTAC.
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