alexa HPLC Separation of Amino Acids is Appropriate | Open Access Journals
ISSN : 2153-2435
Pharmaceutica Analytica Acta
Like us on:
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

HPLC Separation of Amino Acids is Appropriate

Hideharu Shintani*

School of Science, Chuo University, 1-13-27, Kasuga Bunkyo 112-0003, Tokyo, Japan

*Corresponding Author:
Hideharu Shintani
School of Science, Chuo University
1-13-27, Kasuga Bunkyo 112-0003
Tokyo, Japan
Tel: +81425922336
E-mail: [email protected]

Received date: November 16, 2013; Accepted date: November 19, 2013; Published date: November 22, 2013

Citation: Shintani H (2013) HPLC Separation of Amino Acids is Appropriate? Pharmaceut Anal Acta. 4:e158. doi: 10.4172/2153-2435.1000e158

Copyright: © 2013 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 Pharmaceutica Analytica Acta

Concerning amino acid analysis, it is so often used post column derivatization method using ninhydrin or ortho-phthal aldehyde with gradient elution. Ninhydrin and ortho-phtal aldehyde are identical. Problem is that 21 sorts of amino acids cannot always baseline separation (Figure 1).


Figure 1: Several sorts of amino acids are not base-line separation and require at least Rs (resolution) more than 1.5 for baseline separation.

From the Figure, several sorts of amino acids are not base-line separation and require at least Rs (resolution) more than 1.5 for baseline separation. Amino acid chromatographies done by Shimazdu, Hitachi or Nihon Bunko were almost identical. They require baseline separation; otherwise reproducible determination cannot be attained. For that purpose of base-line separation, gradient elution condition must be innovated. Most innovated point is the use of column with higher theoretical plate number. So, I suppose why they do not use capillary-column with higher theoretical plate number.

The above presented chromatogram is representative chromatogram, so analytical validation is further required and gradient condition and elution condition must be innovated to attain base-line separation chromatogram.

The separation condition obtained chromatogram is presented in Table 1.

Time(min) Event Conditions
0.0 Sample injection Na-E buffer,48° C
8.5 Start temp.gradient 48° C to 60° C in 8 min
24.5 Buffer change Na-E to Na-F
41.0 Buffer change Na-F to Na-D
78.0 Reagent pump Ninhydrin to water
79.0 Buffer change Na-D to Na-R
80.0 Buffer change Na-R to Na-E
82.5 Temperature change 60° C to 48°C
84.0 Reagent pump Water to ninhydrin
97.0 Recycle (Start next run)  

Table 1: Standard amino acid analysis.

The elution order of amino acids is from acidic amino acid, neutral amino acid. and then basic amino acid, indicating cation chromatography-column will be used. This selection is normal, but gradient condition is required innovation for base-line separation. Most of acidic amino acid and neutral amino acid are not separated in based-line. This is. further, requires analytical validation to reproducible analytical results.

Select your language of interest to view the total content in your interested language
Post your comment

Share This Article

Recommended Conferences

Article Usage

  • Total views: 11825
  • [From(publication date):
    November-2013 - Sep 26, 2017]
  • Breakdown by view type
  • HTML page views : 8017
  • PDF downloads :3808

Post your comment

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

Peer Reviewed Journals
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri, Food, Aqua and Veterinary Science Journals

Dr. Krish

[email protected]

1-702-714-7001 Extn: 9040

Clinical and Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals


[email protected]

1-702-714-7001Extn: 9042

Chemical Engineering and Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001 Extn: 9040

Earth & Environmental Sciences

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

General Science and Health care Journals

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics and Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001 Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Informatics Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Material Sciences Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Mathematics and Physics Journals

Jim Willison

[email protected]

1-702-714-7001 Extn: 9042

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001 Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

John Behannon

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001 Extn: 9042

© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version