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

Fluorescent Products

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: shintani@mail.hinocatv.ne.jp
Citation: Shintani H (2013) IFluorescent Products. Pharmaceut Anal Acta 4:e152. doi:10.4172/2153-2435.1000e152
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.
Related article at
DownloadPubmed DownloadScholar Google

Visit for more related articles at Pharmaceutica Analytica Acta

Introduction
The reaction of lipid peroxidation products, such as malonaldehyde, with amino groups of proteins, free amino acids, aminophospholipids, or nucleic acid bases, produces fluorescent lipid-peroxidation products known as conjugated Shiff bases with the general structure RN=CHCH=CHNHR’ (N,N’-disubstituted 1-amino- 3-iminopropenes) [1]. When these products are excited at 360 nm the fluorescence maximum is in the region 430-440 nm.
Protocol
Spectrofluorimetric analysis of fluorescent lipid peroxidation products is covered in detail elsewhere [2,3]. The method described here is a simple procedure for detection of lipid-soluble and watersoluble fluorescent products formed during lipid peroxidation of biological membranes.
Lipid-Soluble Fluorescent Products
1. Place membrane sample (e.g. oxidized tissue homogenate, plasma, mitochondria, microsomal suspension, or lipoproteins; 0.5 mL) and ethanol-ether (3:1, v/v, 1.5 mL) in screw-capped tubes (1.3×10 cm, Pyrex).
2. Mix on a vortex mixer for 1 min.
3. Centrifuge for 10 min at 3,000 rpm (4°C).
4. Wash the sediment at the bottom of tube twice with ethanolether (3:1, v/v, 2 mL) repeating assay steps 2 and 3.
5. Pipette the solution (1.5 mL) into a 1-mL quartz cuvette.
6. Measure fluorescence (emission) and excitation spectra by spectrofluorimetry. (The fluorescent lipid-peroxidation products have usually an excitation maximum in the region of 355ー365 nm and an emission maximum at 430-440 nm).
Water-Soluble Fluorescent Products
1. Dissolve sediments from assay of lipid-soluble fluorescent products (above) in 2.0 mL 15% SDS-PBS (phosphate-buffered saline, pH 7.4) on a vortex mixer for 5 min.
2. Centrifuge for 10 min at 3,000 rpm (room temperature).
3. Pipette the solution (1.5 mL) into a 1-mL quartz cuvette.
4. Measure fluorescence (emission) and excitation spectra by spectrofluorimetry.
Calculation
The ftuorescence intensity of quinine sulfate (1 mg mL-1) in H2SO4 solution (0.05M) is the standard for the relative ftuorescence intensity of the sample [2]. Quinine sulfate in the solution has a ftuorescence maximum at 457 nm when excited at 360nm.
Comments
Fluorescent products which bind to proteins are insoluble in common organic solvents and their structures (ftuorophores) can be disrupted, in part, during extraction of lipids with 2:1 (v/v) chloroformmethanol. They are, however, stable in 3:1 (v/v) ethanol-ether, as described elsewhere [4]. It should be noted that the lipid extracts from biological membranes and tissue homogenates contain retinol as one of interfering ftuorescent compounds. Retinol, a lipid-soluble compound, has a ftuorescence maximum at 478nm when excited at 335 nm [5] and its ftuorescence can be removed from the lipid extracts by exposing it to high-intensity ultraviolet light for 30s [2].
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: 11201
  • [From(publication date):
    August-2013 - Aug 29, 2016]
  • Breakdown by view type
  • HTML page views : 7468
  • 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