alexa Eggplant Peel Ethanolic Extract: A Novel and Alternative Stain for Chromosome Banding | OMICS International
ISSN: 2155-9821
Journal of Bioprocessing & Biotechniques

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

Eggplant Peel Ethanolic Extract: A Novel and Alternative Stain for Chromosome Banding

Hayfaa A Al-Shammary1*, Rasha Salh2 and Haidr Sabah2
1Medical Analysis Department, College of Science, University of the Qar, Iraq
2Biology Department, College of Science, University of the Qar, Iraq
Corresponding Author : Hayfaa A Al-Shammary
College of Science Medical Analysis Department
University of The Qar, Iraq
Tel: 078-017-897-65
E-mail:[email protected]
Received October 16, 2014; Accepted January 22, 2015; Published January 26, 2015
Citation: Al-Shammary HA, Salh R, Sabah H (2015) Eggplant Peel Ethanolic Extract: A Novel and Alternative Stain for Chromosome Banding. J Bioprocess Biotech 5:201 doi: 10.4172/2155-9821.1000201
Copyright: © 2015 Al-Shammary HA, et al. 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 Journal of Bioprocessing & Biotechniques

Abstract

Rat and mouse have become important animal models to study various human diseases such as cancer. Here, we use eggplant peel ethanolic extract to visualise chromosome banding. This extract is easy to prepare, a novel, available alternative. The colour of this extract is bright dark green colour. This stain gave us good result for show the chromosomes banding, we used Thin Layer Chromatography (TLC) examination, Infrared (IR) spectroscopy and the spectrum ultraviolet UV to investigate about the compound and functional groups in this extract. Use of this extract represents a new method for chromosome banding and a viable alternative to existing chromosome staining methods.

Keywords
Ethanolic extract; Eggplant; Chromosomes bandings
Introduction
A wide variety of stains are used to visualise chromosomes under the microscope. The aceto-orcein, acetocarmine, gentian violet, and haematoxylin readily stain chromatin are classical cytological stains allowing for easy visualisation of chromosomes under a standard light microscope [1].
Numerous methods are available for identifying chromosomes and preparing karyotypes for clinical and research purposes. The most common methods of dye-based chromosome banding are Giemsa- (G), reverse- (R), centromere- (C), and quinacrine- (Q) banding. Q- and G-banding introduced a new era in which individual chromosomes could be definitively identified. With this capability, it also became possible to localize regions of variable size and staining to specific chromosomes. In particular, Q- and C-banding can reveal distinct classes of heteromorphisms that are not necessarily detectable in non-banded chromosomes. The most distinctive heteromorphism revealed by Q-banding was the brightly fluorescent distal long arm of the Y chromosome. The size of this brightly fluorescent segment varies from being almost negligible in size to being the longest segment on the Y long arm. Q-banding also revealed variations in staining of chromosomes 3, 4, 13-15, and 21-22 of the human karyotype [2- 6]. Although G-banding techniques have become widely used for chromosome identification, C-banding also revealed size variations of heterochromatin around the centromeres of every chromosome; these could be more easily quantified in banded than in non-banded chromosomes. The heterochromatin regions of chromosomes 1, 9, and 16, and in the distal long arm of the Y, evident in non-banded chromosomes, were especially visible by C-banding [6-10].Since banding may be a reflection of the difference in the structure along the length of a chromosome, studying the mechanism of banding can improve our understanding of chromosome structure [11-13]. Banding can be used for chromosome identification (karyotyping), and for identifying abnormalities of chromosome number, translocations of material from one chromosome to another, and deletions, inversions or amplifications of chromosomes. Q-banding was discovered by Caspersson et al. [2] who applied quinacrine mustard dihydrochloride (QM) staining to human chromosome X. This resulted in the discovery that the end of the long arm of the Y chromosome was brightly fluorescent: bright enough that the human Y chromosome could be easily detected in interphase as well as in metaphase cells [14].
G-banding is the most widely used banding technique in clinical laboratories. The G-banding method uses acid fixation with saline treatment, followed by Giemsa staining [15]. Application of proteolytic enzymes such as trypsin [16,17] or pancreatin [18,19] improved the banding pattern. Dark-staining G-bands indicate AT-rich regions of chromosomal DNA that are more condensed replicate their DNA later than less condensed GC-rich regions [20].
R-banding was discovered by Utrillaux and Lejeune [21]. Due to technical difficulties and fluorescent requirements, it is not a widely used method of banding.
The observation by Pardue and Gall that the centromeres of mouse chromosomes stained darker than other chromosomal regions led to the discovery of C-banding [22]. Arrighi and Hsu [23,24] developed a modified technique C-banding by applying Giemsa staining; other modified versions of C-banding have since been developed.
The technique Cd-banding was described by Eiberg [25]. The G-11 technique and finally Silver Staining was developed by Howell and Black [27].
Eggplant, also called aubergine or brinjal (Solanum melongena L.) is an edible fruit, which is cultivated globally, but particularly in Asia and Europe [28,29]. Two kinds of anthocyanin have been isolated and identified from the purple cultivar [30,31]. The role of anthocyanin pigments as medicinal agents is well-accepted dogma in folk medicine throughout the world; indeed, eggplant is used in traditional medicines [30]. The anthocyanin delphinidin-3-rutinoside was identified from eggplant by HPLC-DAD-MS3 analyses [32]. A second anthocyanin, delphinidin-3-(p-coumaroylrutinoside)-5-glucoside (nasunin), was isolated as purple coloured crystals from eggplant peels [33].
Materials and Methods
Ethanolic extract
Eggplant peel was removed and dried in the shied. 25 grams of the resulting powder was extracted in 250 ml 96% ethanol alcohol by using the socholite for 6 hours. The colour of this extract is bright dark green colour.
Thin layer chromatography (TLC) examination
TLC was carried out using ethyl acetate: hexane (1: 9) solvent. Chemical analysis with ethyl acetate: hexane (6: 4) (1: 9) determined the presence of one separated compound (Figure 1).
Infrared (IR) spectroscopy
IR spectroscopy (250-4500 cm) indicated the presence of C-O, C=O, C=C, C-H and CHO=O bonds, and the presence of benzene ring (Figure 2).
The spectrum ultraviolet UV of Eggplant ethanolic extract
Scan spectrum curve show the presence of absorption at the wavelength packages (max = 200.00-780.00 nm) has shown in Figure 3
Cytogenetic examinations
Rat chromosomes were prepared for cytogenetic examination. The animals were sacrificed and chromosome aberrations relieved using the method of Giri et al. [34]. Prepared slides were rinsed in ammonium alum solution for 2-3 minutes and stained for 15 min in an eggplant alcohol extraction solution. The staining solution was filtered immediately before use. Stained slides were air-dried before being examined and photographed under a light microscope.
Results
As shown in Figure 4, stained chromosomes, with banding, could be clearly visualised after treatment with eggplant peel ethanolic extract, and Figure 5 which stained with Gemza shows no banding.
Discussion
Chromosome staining is used to enhance the contrast between different cellular components. Each chromosome arm is divided into regions, or cytogenetic bands, that can be seen using a microscope and special stains. The cytogenetic bands are labelled p1, p2, p3, q1, q2, q3, etc., counting from the centromere out toward the telomeres. These bands provide further information about the chromosomes. Since each chromosome number produces unique bands, this method can be used to identify individual chromosomes [35].
The result of this study showed that staining with eggplant alcohol extract is a viable method of staining rat chromosomes, and produced visible banding regions. In normal chromosomal stain procedure, metaphase chromosomes are treated with trypsin and stained with Giemsa stain. The major bands regions in chromosome are constitutive heterochromatin, facultative heterochromatin, and euchromatin. Heterochromatic regions, which tend to be ATrich and relatively gene-poor, stain more darkly in G-banding. In contrast, less condensed chromatin, which tends to be GC-rich and more transcriptionally active, incorporates less Giemsa stain, and hence these regions appear as light bands in G-banding. In this study ammonium alum solution has been used. This compound may bind to the DNA through intercalation, minor or major groove binding, or external binding, and thus have an effect on the nucleotides in the DNA. The mode of binding depends on the nature of the interaction between the stain and the DNA, which may be either covalent or noncovalent [35].
Using IR spectroscopy (IR ), we detected the presence of functional groups including C-O, C=O, C=C ,C-H, COH=O and benzene ring . This range of functional groups could produce more than one type of DNA-stain interaction, suggesting that our eggplant alcohol extract stain can bind to DNA through several different binding modes [36]. The type of bands produced may also depend on the extent of denaturation undergone by the chromosome structure.
Acknowledgements
We would like to thank the college of science from the University of The Qar for facilitates and our families for the support they gave us.
References

Figures at a glance

 

Figure Figure Figure Figure Figure
Figure 1 Figure 2 Figure 3 Figure 4 Figure 5
Select your language of interest to view the total content in your interested language
Post your comment

Share This Article

Relevant Topics

Article Usage

  • Total views: 12680
  • [From(publication date):
    February-2015 - Nov 15, 2018]
  • Breakdown by view type
  • HTML page views : 8836
  • PDF downloads : 3844
 

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 2018-19
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri and 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- 2018 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version