Investigation of Hypericum Scabrum L.'s Methanol Extract with Disposable Electrochemical DNA Biosensors.

Abstract

In this study, a voltammetric sensor was used to determine the interactions of the methanol extract of the plant Hypericum scabrum L., with the nucleic acid molecule "DNA". Hypericum scabrum L., which has high antioxidant activity is also used in the treatment of diseases such as ulcers, constipation, hemorrhoids, rheumatism, and menstrual diseases. Our biosensor system has been designed using differential pulse voltammetry (DPV) and cyclic voltammetry (CV) methods. In the study; A 3-electrode system consisting of a pencil graphite electrode, auxiliary electrode, and reference electrode will be used. In the study, firstly, an electrochemical analysis of the plant extract was performed and electroactivity was determined. In the second part, the single-stranded (ssDNA) and double-stranded (dsDNA) DNA structures were attached to the electrode surface and then, the DNA-modified electrodes were interacted with Hypericum scabrum's methanol extract solutions. To determine the effect of the bioactive substances contained in the extract on DNA structures, the change in guanine oxidation signal, which is one of the most active bases of DNA and is oxidized at approximately +1.0 V, was measured account before and after the interaction. After the analysis of synthetic DNA fragments was completed, the effect of H. scabrum's methanol extract on DNA was analyzed in a short time and in a very simple way with the designed electrochemical biosensor. As a result of the studies, the differences between the responses of the bioactive substances contained in the plant and the differences in the oxidation response of the guanine base were determined using the DPV technique. In the determination, whether the flavonoids [1, 2], which are thought to be included in the methanol extract, have an electrochemical response (whether they are active or not) was investigated using the cyclic voltammetry technique (CV), one of the electrochemical determination methods. Differences in experimental parameters (environment and pH, substance concentration, DNA concentration, etc.) were examined with the DPV technique.

Keywords

• Hypericum scabrum
• DNA
• Biosensors
• Electrochemistry

Hypericum scabrum L.'nin Metanol Ekstresinin Tek Kullanımlık Elektrokimyasal DNA Biyosensörleri ile İncelenmesi.

Öz

Bu çalışmada, halk arasında ülser, konstipasyon, hemoroit, romatizma ve menstrüal hastalıklar gibi rahatsızlıkların tedavisinde kullanılan, yüksek antioksidan aktiviteye sahip Hypericum scabrum L. bitkisine ait metanol ekstresinin nükleik asit molekülü “DNA” ile etkileşimlerinin tayini için Diferansiyel Puls Voltametrisi (DPV) ve Dönüşümlü Voltametri (CV) yöntemleri kullanılarak biyosensör sistemleri tasarlanmıştır. Çalışmada; kalem grafit elektrot, yardımcı elektrot ve referans elektrottan oluşan 3’lü elektrot sistemi kullanılacaktır. Çalışmada ilk olarak bitki ekstresinin elektrokimyasal incelenmesi yapılarak elektroaktivite tayini sağlanmıştır. İkinci kısımda elektrot yüzeyine tutturulan tek zincirli (ssDNA) ve çift zincirli (dsDNA) DNA yapıları ile bitkiden hazırlanan metanol ekstresinin etkileşimi sağlanmıştır. Ekstrede yer alan biyoaktif maddelerin DNA yapılarına etkisinin tayini için, etkileşim öncesi ve sonrasında DNA’nın en aktif bazlarından biri olan ve yaklaşık +1,0 V’da yükseltgenen guanin bazı sinyallerindeki değişim dikkate alınmıştır. Sentetik DNA parçalarının analizleri tamamlandıktan sonra H. scabrum metanol ekstresinin DNA üzerine olan etkisi tasarlanan elektrokimyasal biyosensör ile kısa sürede ve oldukça basit bir şekilde analizlenmiştir. Yapılan çalışmalar sonucunda bitki içeriğinde yer alan biyoaktif maddelerin verdiği yanıtlar arasındaki farklılıklar ve guanin bazının yükseltgenme yanıtındaki farklılıklar DPV tekniği kullanılarak tayin edilmiştir. Tayinde ayrıca metanol ekstresinin yer aldığı düşünülen flavonoitlerin [1, 2] elektrokimyasal olarak yanıtlarının olup olmadığı (aktif olup olmadıkları) elektrokimyasal tayin yöntemlerinden dönüşümlü voltametri tekniği (CV) kullanılarak araştırılmıştır. Deneysel parametrelerdeki farklılıkların (ortam ve pH, madde konsantrasyonu, DNA derişimi vb.) DPV tekniğiyle incelenmesi yapılmıştır.

Anahtar Kelimeler

• Biosensör
• Hypericum scabrum
• DNA
• Elektrokimya

Kaynakça

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