alexa HPLC Analysis of Vitamin A and Carotenoids | OMICS International
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 Analysis of Vitamin A and Carotenoids

Hideharu Shintani*

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

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

Received date: January 28, 2013; Accepted date: March 15, 2013; Published date: March 20, 2013

Citation:Shintani H (2013) HPLC Analysis of Vitamin A and Carotenoids. Pharmaceut Anal Acta 4:218. doi: 10.4172/2153-2435.1000218

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

Introduction

Retinol and carotenoids are extracted from plasma with ethanol containing butyl-hydroxytoluene (BHT). Retinol is determined by HPLC with spectrofiuorimetric detection (λex 340 nm; λcm 460 nm). Carotenoids are also determined by HPLC with UV detection (A453 nm) or with an electrochemical detector (ECD, 900mV). Figure 1 shows a typical chromatogram from human blood plasma carotenoids [1,2].

pharmaceutica-analytica-acta-Typical-HPLC-chromatogram

Figure 1: Typical HPLC chromatogram of carotenoids from blood plasma Detection was by ECD at 900 mV.

Protocol

1. Add BHT in ethanol (0.015%, 1 ml) to plasma (0.2 ml) and shake.

2. Add n-hexane (5 ml) and shake.

3. Centrifuge for 10 min at 3000 rpm.

4. Evaporate upper layer (4 ml) under N2 gas.

5. Add ethanol (100 ml) and analyse by HPLC.

HPLC was performed with an lrica Instruments (Kyoto, Japan) Σ-871 chromatograph equipped with a 250 mm × 4 mm i.d. lrica RP- 18 column and fluorescence detection (λex 340 nm; λem 460 nm). The mobile phase was 95:5 (v/v) ethanoi-H2O at a flow rate of 0.7 ml/min. The external standard was retinal prepared from retinyl palmitate

Discussion

ECD is highly sensitive and is used when small amounts of biological sample are available. The more the applied voltage increased, the lesser the selectivity. However, the sensitivity increased with increased applied voltage. ECD is in general 10 to 100 times more sensitive than UV at 220 nm. The sensitivity of fluorescence detection was almost identical to ECD.

In figure 1 separation of the interest compounds from the admixtures must be further improved. For that purpose, change of constituent of the mobile phase, selection of column, gradient elution and solid phase extraction must be considered. These studies may result in appropriate separation.

References

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: 12220
  • [From(publication date):
    March-2013 - Dec 12, 2017]
  • Breakdown by view type
  • HTML page views : 8382
  • PDF downloads : 3838
 

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 & Aquaculture Journals

Dr. Krish

[email protected]

1-702-714-7001Extn: 9040

Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001Extn: 9040

Clinical Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

Food & Nutrition Journals

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

General Science

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics & Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Materials Science Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Nursing & Health Care Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

Ann Jose

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001Extn: 9042

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