Penggunaan darah kering untuk pengukuran asam lemak tran

Penggunaan darah kering untuk pengukuran asam lemak tran

Latar Belakang

Pengukuran asam lemak asam lemak trans khususnya telah memperoleh kepentingan dalam beberapa kali. Di antara biomarker tersedia berbagai, jaringan adiposa dianggap yang terbaik untuk asupan makanan jangka panjang tetapi sifat invasif aspirasi jaringan mengurangi utilitas. Proses mengeluarkan darah merupakan metode yang jauh kurang invasif koleksi sampel ketika sejumlah besar peserta yang terlibat dalam studi ini dan karena itu alternatif, paling cocok untuk studi populasi yang besar berbasis. Dalam penelitian ini asam lemak (dengan penekanan khusus pada asam lemak trans) ekstraksi dari darah melihat dan dikeringkan pada kertas filter dilakukan untuk menyederhanakan prosedur sampel pengumpulan dan transportasi.

Metode

Sampel darah diambil dari 19 relawan yang sehat. Darah itu terlihat (30 titik dari 10 ml masing-masing) pada kertas filter, dikeringkan pada suhu kamar dan disimpan pada suhu 4 ° C dalam zip-lock tas poli. Untuk darah perbandingan seluruh disimpan pada -70 ° C secara bersamaan dianalisis.

Hasil
Sebuah perjanjian yang baik terlihat antara trans nilai asam lemak yang diperoleh dalam darah kering dan seluruh darah yang terlihat dari koefisien korelasi pearson (‘r’ untuk monounsaturated (trans) 0.70 dan polyunsaturated (trans) 0,692 masing-masing). Korelasi intrakelas koefisien untuk trans tak jenuh tunggal adalah 0,805 dan untuk trans polyunsarurated adalah 0,776.

Kesimpulan
Bercak darah kering dapat digunakan untuk analisis asam lemak trans.

Drug Development Services: Dried Blood Spot (DBS)

BASi has emphasized the importance of reducing animal numbers and refining data. We are continuing this tradition as we move forward with utilizing Dried Blood Spot (DBS) technologies and have a number of initiatives that capture the advantages dried sample spots offer.

Dried Blood Spot sampling offers advantages for the collection, storage and shipping of blood samples for drug analysis. The small volumes utilized in DBS are a good fit for multiple small volume collections from animals such as mice and rats. Use of DBS technologies for this purpose make it possible to reduce the overall number of experimental animals required for development. However, this collection technique goes beyond early discovery and can be a low-stress sampling option for mice as well as man.

Bioanalytical DBS Services

• Collaborative efforts between our bioanalytical labs in the US and the UK have allowed us to successfully evaluate the bioanalysis of dried blood spots and have developed DBS analysis expertise at each of our three bioanalytical facilities.
• Our staff and programs are flexible to generate both the study design and timelines you need.
• We offer complementary services in preclinical development, toxicology, bioanalysis, and TK reporting.

Culex® Automated DBS Sampling

• BASi is the designer and manufacturer of theCulex^® Family of instruments for automated blood sampling and has designed a solution to integrate DBS into the blood collection system.
• Culex^® will collect accurate and reliable Dried Blood Spot samples at user-programmed time points for direct delivery onto DBS paper.
• Our unique sample collection card allows drawing of up to 32 DBS samples with 1-4 samples per time point and the ability to retain separate replicate samples.
• Culex^® reduces human intervention during sample collection, resulting in reduced stress when compared to sampling manually.
  

The BASi Advantage

• Integration of DBS sampling on Culex^® for small and large animal models complements our existing DBS bioanalytical capability.
• The combination of our in-house team of scientists and engineers ensures we will continue to make improvements to the collection and analysis of Dried Blood Spot samples.
• We provide our customers with DBS solutions that encompass the drug development process from discovery to clinical trials.

Related Articles

• The Culex® Automated In Vivo Sampling System: A Novel Technique for Dried Blood Spot (DBS) Collection in Preclinical Studies
  Peter White, David Hopper, Robin Lane, Tyler McGraw, Bhavna Mistry, Amanda Plumb and Candace Rohde
   
• Validation of an LC-MS/MS Method for the Determination of Quetiapine, Norquetiapine and Quetiapine Sulphoxide in Human Whole Blood Using the Dried Blood Spot TechniqueNeil Doleman, Robin Lane
  BASi, UK


• Evaluation of the Dried Blood Spot Technique for the Quantification of Diltiazem in Human Blood
  Neil Doleman, BASi, UK

SOURCE:https://www.basinc.com/services/DBS.html

WHAT IS DRIED BLOOD SPOT SAMPLING ?

Dried Blood Spot (DBS) sampling is an easy and inexpensive way of collecting, shipping and storing blood samples. It represents an alternative to the body fluid based method (plasma, liquid blood or serum). Originally developed for the purpose of neonatal screening, it has been around for almost 50 years. As a microsampling technique it has gained increasing importance since this method offers strong advantages compared to the conventional collection and analysis of blood or plasma samples.

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These advantages include the need for remarkably lower blood volumes and easier shipping and storage, often at ambient temperatures. This leads to a simplification of the blood collection process and a reduction of the costs involved.
For DBS analysis, blood samples are spotted onto commercially available DBS cards, dried and shipped to the laboratory for mass spectrometry (MS) analysis. But prior to MS analysis, the blood sample needs to be extracted from the DBS card. Until now, this represented a major drawback of the technique as DBS sample extraction required several manually performed process steps, and therefore was time consuming and costly. The fully automated CAMAG DBS-MS 500 extraction system makes DBS analysis remarkably easier and offers the efficient alternative compared to conventional extraction techniques.
Furthermore, in preclinical studies the number of test animals can be reduced (due to the need for lower blood volumes) which is in accordance with the 3R requirement of animal studies (replacement, reduction, refinement).

SOURCE:http://www.camag.com/en/dbs/what_is_dried_blood_spot_sampling.cfm

Dried blood spot testing (DBS)

Dried blood spot testing (DBS) is a form of biosampling where blood samples are blotted and dried on filter paper. The dried samples can easily be shipped to an analytical laboratory and analysed using various methods such as DNA amplification or HPLC.

History

The concept that capillary blood, obtained from pricking the heel or finger and blotted onto filter paper, could be 

used to screen for metabolic diseases in large populations of neonates was introduced in Scotland by Robert 

Guthrie in 1963. Neonatal screening for phenylketonuria became nationwide in 1969-70. Since then, Guthrie card 

samples have been collected routinely from infants in over 20 countries to screen for phenylketonuria and more 

recently for congenital hypothyroidism, sickle cell disorders and HIV infection. The limitations of sensitivity and 

specificity when screening such small volumes of blood restricted the use of dried blood spots for many years. 

However, recent advances such as the production of monoclonal antibodies, expression of synthetic proteins, and

 the introduction of the polymerase chain reaction have overcome many of these problems.

This type of blood testing is now available for use at home by consumers in the U.S. Available blood tests include

 vitamin D, estrogen, testosterone, cortisol, TSH and lipids. New York is the only state that prohibits home blood

 spot testing.

Procedure

Dried blood spot specimens are collected by applying a few drops of blood, drawn by lancet from the finger, heel or

 toe, onto specially manufactured absorbent filter paper. The blood is allowed to thoroughly saturate the paper and

is air dried for several hours.^[Specimens are stored in low gas-permeability plastic bags with desiccant added to 

reduce humidity, and may be kept at ambient temperature, even in tropical climates.

Once in the laboratory, technicians separate a small disc of saturated paper from the sheet using an automated or

 manual hole punch, dropping the disc into a flat bottomed microtitre plate. The blood is eluted out in phosphate 

buffered saline containing 0.05% Tween 80 and 0.005% sodium azide, overnight at 4°C. The resultant plate 

containing the eluates forms the "master" from which dilutions can be made for subsequent testing.

As an alternative to punching out a paper disc, recent automation solutions extract the sample by flushing an

 eluent through the filter without punching it out.

Dried blood spot testing for HIV infection

The technology holds promise for expanding diagnostic services to HIV-infected infants in resource-poor settings 

due to the samples' longer lifespan with reduced need for refrigeration and the less invasive nature of the test 

compared with other methods. Unlike ELISA testing for HIV-antibodies in the blood, which may be transmitted to

 infants in pregnancy independently of the virus itself, dried blood spot testing can be used to detect genetic 

material of the actual virus, thereby avoiding the likelihood of a false positive result. DBS specimens also pose less

 of a biohazard risk to handlers, and are easier to transport or store than liquid blood specimens.

Principle

The reason for stability of DNA, RNA or protein could be attributed to the fact that the biological material binds to 

the matrix of the filter paper and the process of drying excludes water which is an important factor necessary for 

protease or nuclease to act. Binding of the biological material also binds several inhibitors which may interfere with

 various nucleic acid amplification methods.

SOURCE: http://en.wikipedia.org/wiki/Dried_blood_spot