ISSN: 2167-7689
Pharmaceutical Regulatory Affairs: Open Access
Make the best use of Scientific Research and information from our 700+ peer reviewed, Open Access Journals that operates with the help of 50,000+ Editorial Board Members and esteemed reviewers and 1000+ Scientific associations in Medical, Clinical, Pharmaceutical, Engineering, Technology and Management Fields.
 
Meet Inspiring Speakers and Experts at our 3000+ Global Conferenceseries Events with over 600+ Conferences, 1200+ Symposiums and 1200+ Workshops on
Medical, Pharma, Engineering, Science, Technology and Business

Immobilized Enzyme Column Combined with HPLC and Column Switching Method for the Analysis of Complicated Matrix Such As Body Fluids

Hideharu Shintani*
Faculty of Science and Engineering, Chuo University, Tokyo, Japan
Corresponding Author : Hideharu Shintani
Faculty of Science and Engineering
Chuo University, 1-13-27, Kasuga, Bunkyo
112-8551, Tokyo, Japan
Tel: +81425922336
Fax: +81425922336
E-mail: shintani@mail.hinocatv.ne.jp
Received October 10, 2014; Accepted October 11, 2014; Published October 14, 2014
Citation: Shintani H (2014) Immobilized Enzyme Column Combined with HPLC and Column Switching Method for the Analysis of Complicated Matrix Such As Body Fluids. Pharmaceut Reg Affairs 3:e142. doi: 10.4172/2167-7689.1000e142
Copyright: © 2014 Shintani H. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Visit for more related articles at Pharmaceutical Regulatory Affairs: Open Access

Enzymes are bio-catalysts which affect transformation of substrates to products with high specificity. The usage of enzymes in domestic and industrial applications is well known and has been well documented since the early history of civilization. With the advances in understanding of enzymology, usage of enzymes in industrial and biotechnological processes and molecular medicine has proliferated.
One of the key factors in the widespread application of enzymes in modern technologies is the development of enzyme immobilization techniques (Figure 1), which overcome certain practica1, functional and economic constraints. Many natural enzymes can be stabilized by immobilization on solid matrices, with most of the activity retained for a variety of applications. An important application of immobilized enzymes is in liquid chromatography. In the last decade, post-column enzyme detection has become established as an important discipline in liquid chromatography. The new detection approach offers more sensitive and specific ways for measuring major classes of biomolecules (Figure 2).
Reactors are fabricated by packing the immobilized enzymes into small columns, which can be placed immediately after or prior to an HPLC column. When an analyte, which is a natural substrate of the enzyme, passes through the reactor, it will be transformed into a product in the presence of a coenzyme. The enzymatic reaction can be designed in such a way that one of the reaction products will be amenable to optical (UV or visible) or electro-chemical detection (Figure 3). Hence、compounds that are otherwise difficult to detect by conventional approaches are rendered readily detectable with improved sensitivity. Alternatively, certain reactors can be placed immediately before the injector to transform an analyte into a product that can be chromatographed for subsequent detection. The specificity of the reactor, in general, is dependent on the nature of the immobilized enzymes. Some enzymes have broad specificity, catalysing the transformation of a specific functionality of an entire family of compounds、while other enzymes have absolute specificity, reacting with a single substrate (Figure 4).
The enzyme reaction detection liquid chromatography systems offer new bio-analytical techniques to pharmaceutical scientists and biochemists for effective and sensitive measurement of drugs and conjugated metabolites in pharmaceutical formulations and physiological metabolites. They also offer the broader analytical community a new technique with great potential [1] (Figure 5).
This editorial will describe the analytical application of immobilized enzyme reactors posterior to [1] or prior to HPLC [2] for selective analysis of specific compound in the complicated matrix. The integration of the reactors into detection systems in liquid chromatography provides new ways of measuring biomolecules with very high sensitivities. In this editorial one example of urea analysis combined with immobilized urease and indophenol or OPA detection are provided [1,2]. Immobilized urease can be available commercially and this procedure is also presented as Figure 1. This information may help to prepare immobilized enzymes to the researchers who are studying for the analysis in the complicated matrix.
References
Select your language of interest to view the total content in your interested language
Post your comment

Share This Article

Relevant Topics

Recommended Conferences

Article Usage

  • Total views: 11183
  • [From(publication date):
    December-2014 - Jul 26, 2016]
  • Breakdown by view type
  • HTML page views : 7450
  • PDF downloads :3733
 
 

Post your comment

captcha   Reload  Can't read the image? click here to refresh

OMICS International Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
 
 
OMICS International Conferences 2016-17
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings
 
 

Contact Us

Agri, Food, Aqua and Veterinary Science Journals

Dr. Krish

agrifoodaquavet@omicsinc.com

1-702-714-7001 Extn: 9040

Clinical and Biochemistry Journals

Datta A

clinical_biochem@omicsinc.com

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

business@omicsinc.com

1-702-714-7001Extn: 9042

Chemical Engineering and Chemistry Journals

Gabriel Shaw

chemicaleng_chemistry@omicsinc.com

1-702-714-7001 Extn: 9040

Earth & Environmental Sciences

Katie Wilson

environmentalsci@omicsinc.com

1-702-714-7001Extn: 9042

Engineering Journals

James Franklin

engineering@omicsinc.com

1-702-714-7001Extn: 9042

General Science and Health care Journals

Andrea Jason

generalsci_healthcare@omicsinc.com

1-702-714-7001Extn: 9043

Genetics and Molecular Biology Journals

Anna Melissa

genetics_molbio@omicsinc.com

1-702-714-7001 Extn: 9006

Immunology & Microbiology Journals

David Gorantl

immuno_microbio@omicsinc.com

1-702-714-7001Extn: 9014

Informatics Journals

Stephanie Skinner

omics@omicsinc.com

1-702-714-7001Extn: 9039

Material Sciences Journals

Rachle Green

materialsci@omicsinc.com

1-702-714-7001Extn: 9039

Mathematics and Physics Journals

Jim Willison

mathematics_physics@omicsinc.com

1-702-714-7001 Extn: 9042

Medical Journals

Nimmi Anna

medical@omicsinc.com

1-702-714-7001 Extn: 9038

Neuroscience & Psychology Journals

Nathan T

neuro_psychology@omicsinc.com

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

John Behannon

pharma@omicsinc.com

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

social_politicalsci@omicsinc.com

1-702-714-7001 Extn: 9042

 
© 2008-2016 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version