Türkiye'de farklı yörelerde yetişen vişne meyve ekstraktlarında polifenol kapasitesi ve antosiyanin profilinin karşılaştırılması
Poliphenol content and antioxidant profile of sour cherries (Prunus cerasus L.) from Turkey
Yazar: KÜBRA AYDINLI
Danışman: DOÇ. DR. ESRA ÇAPANOĞLU GÜVEN
İstanbul Teknik Üniversitesi / Fen Bilimleri Enstitüsü / Gıda Mühendisliği Ana Bilim Dalı / Gıda Mühendisliği Bilim Dalı
Derece: Yüksek Lisans, 2018
Konu: Gıda Mühendisliği = Food Engineering

Özet:
Bu çalışma kapsamında farklı yörelerde yetişen vişne (Prunus cerasus L.) meyvesinde doğal antosiyanin düzeyi ve dağılımı ile antioksidan ve polifenol kapasitesinin fermentasyon prosesinin etkisi ile değişimlerinin belirlenmesi, polifenol ve antosiyanin düzeyi ile antioksidan kapasitesi arasındaki ilişkinin ortaya konulması amaçlanmıştır.
Türkiye'de başlıca yetişme yöresi olan Akşehir, Çay, Doğrugöz ve Şaphane olmak üzere 4 farklı yöreye ait vişne örneğinin analizleri yapılmıştır. Analiz materyali olarak kullanılan meyve ekstraktları 3 tekrarlı olarak temin edilmiş ve her grup örneğin, antioksidan bileşenleri belirlenmiştir. Meyvelerin toplam fenolik madde içerikleri, Folin-Ciocalteu yöntemiyle belirlenmiştir. Toplam flavonoid madde tayini kolorimetrik yöntemle yapılmış, antosiyanin tayini için de pH diferansiyel yöntemi kullanılmıştır. Antioksidan aktivite tayini için DPPH (2,2- Difenil-1-Pikrilhidrazil), ABTS (2,2'-azino-bis(3-etilbenztiazolin-6-sülfonik asit)), ve CUPRAC (Bakır (II) İndirgen Antioksidan kapasitesi) yöntemleri kullanılmıştır. Vişne meyvesinde bulunan başlıca fenolik madde ve antosiyanin fraksiyonlarını HPLC kromatogramında belirlemek adına 280, 312, 360 ve 512 nm dalga boylarında çalışılmıştır. 
Yapılan spektrofotometrik analizler ve HPLC analizinin çıktısı olan tüm sonuçlar, SPSS ile istatistiki olarak yorumlanmıştır. Polifenol ve antosiyanin profili ve kapasitesinin yöre, fermentasyon, yöre*fermentasyona göre anlamlı farklı olup olmadığı değerlendirilmiştir.
Ayrıca başlıca antosiyanin fraksiyonları olarak siyanidin-3-glukozit, siyanidin-3-rutinozit tanımlanmış ve nicelleştirilmiştir. Bunun yanı sıra, protokateşik asit, kateşin, epikateşin, neoklorojenik asit, kafeik asit, p-kumarik asit, kuersetin-3-rutin, kuersetin-3-glikozit tanımlanmıştır.

Abstract:
In this study, it was aimed to determine the correlation between the level and fraction of natural anthocyanin and the effect of fermentation process of antioxidant and polyphenol capacity, polyphenol and anthocyanin levels and antioxidant capacity in cherry juice grown in different regions of sour cherry (Prunus cerasus L.). Nowadays, with the increase in the awareness about the effect of the consumers on the health effects of the naturally occurring components in the content of foods, studies related to the phenolic components of the antioxidant properties have increased.

Cherry (Prunus cerasus L.) is cultivated in many countries such as Russia, Poland, Ukraine, USA. The production capacity were examined for 2016, Turkey takes place 3rd in the world with 192.500 mt / ton, produces 14% of the cherries of volume of production. When sour and sweet cherries considered together, Turkey is the largest producer in the world. The main export markets are the EU and Russia. According to the National Agricultural Statics Service, 99% of the worldwide cherry is consumed as processed fruit such as fruit juice, jam or canned sour cherry.

In recent years, the positive effects of various components in the composition of fruits and vegetables on health have attracted more attention. The increase in consumption of fruits and vegetables rich in antioxidant has gained more importance with the increase in awareness about regular and healthy nutrition. From past to present, fruits and vegetables consumed as thesis can be processed in fruit juice and canned food or consumed by freezing and drying. Like every fruit, cherry has its own anthocyanin distribution, and its antioxidant capacity, such as the color of cherry, is largely due to these compounds. Therefore, these compounds should be protected as much as possible in terms of both quality and health. However, these compounds can rapidly change form during process and storage and decrease bioavailability and bioactivity. Therefore, it is important to determine whether the anthocyanins are affected by the processes applied and to determine the conditions accordingly.

Cherry fruit contains significant amounts of phenolic antioxidants, especially anthocyanins. There are also other groups such as hydroxycinnamates, flavonols and flavan-3-oller (procyanidins). There are many factors that effect on the phenolic content and antioxidant capacity of cherry. The results obtained from various studies show that the composition and content of the phenolic compounds in cherries are affected by their species, growing season and growing location

Legumes, grains, vegetables, plants and fruits are greatly affected by phytochemicals and their bioavailability and fermentation. This leads to an increase in the properties of antioxidant components, which may be useful for the treatment and / or prevention of diseases such as cancer. Therefore, fermentation, especially with probiotics, is considered to be an important process not only for preservation, but also for the production of functional foods enriched with bioactive and antioxidant properties. 

As research material, 4 different varieties of cherries obtained from main producing regions in Turkey. This regions are Çay Region, Şaphane Region, Akşehir Region and Doğrugöz Region.  The sour cherry samples have been treated with S. cerevisiae S1 and S. cerevisiae S2 strains were treated 4 times at water and ethanol at room temperature. For extraction the solvent was obtained by evaporation of the solvent in a spray dryer under vacuum. These samples were supplied as an extract. The strains of S. cerevisiae were two and the strains were coded as S1 and S2. The brix value of the cherry fruit extracts varies between 65-70%.
The analytical properties anlaysed in fruit extracts in 3 replicates and the antioxidant and polyphenolic compounds were determined. Total phenolic content has been determined by Folin-Ciocalteu method and for antioxidant capacity, DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-Azino-di- [3-ethylbenzthiazoline sulfonate (6)]), and CUPRAC (Cupric reducing antioxidant capacity) methods have been used. Anthocyanin determinations were carried out by pH differential method and Flavonoid assays were carried out according to colorimetric assays. In order to determine the major phenolic substances and anthocyanin fractions present in cherry juice on HPLC chromatograms, anlayses conducted at wave lengths of 280, 312, 360 and 512 nm.

All the results are interpreted as statistical by SPSS. Post hoc analysis was performed by using SPSS program and the results were interpreted in confidence interval (p <0.01) and (p <0.05). It was evaluated whether the profile and capacity of polyphenol and anthocyanin were significantly different from region, fermentation, region*fermentation.

It was found that there was a high positive correlation between the total flavonoids determination, DPPH and CUPRAC with total phenolics determination and an negative correlation with the determination of anthocyanin. In the total antioxidant capacity analyzes, the highest values were obtained in CUPRAC analysis. Differentiation of values in DPPH and ABTS methods has been supported by literature. For each analysis parameter, whether there was a significant difference between the region, fermentation and local fermentation variables, significant differences have been observed in the studies. According to the total phenolic substance and flavonoid content, Akşehir region has been differentiated from all regions. Sub-groups generated for each parameter were reported in Annex B. When the effects of S1 and S2 strains were examined in total phenol, total flavonoid, CUPRAC and ABTS analyzes, it was observed that S2 strains increased the capacity.

In the HPLC analysis, 280, 312, 360 and 512 nm absorbance values were studied. The reason for the study of these wavelengths; When the literature is examined, it is stated that the polyphenols in the cherry samples give their max absorbance values at these wavelengths. When evaluating the obtained chromatograms, the maximum wavelength has been selected and reflected on the results. Cyanidin-3-glucoside, cyanidin-3-rutinoside has also been identified and quantified as major fractions of anthocyanins. In addition, protocatechic acid, catechin, epicatechin, neochlorogenic acid, caffeic acid, p-coumaric acid, quercetin-3-rutin, quercetin-3-glucoside have been identified. In addition, the correlation between the results of the monomeric total anthocyanin measurement and the results obtained from HPLC revealed a positive positive correlation (p <0.01).

The region and fermentation effects on antioxiadant capacity and/or fraction were investigated and reported. Although the variability between samples has been demonstrated, it has been possible to make a comparison to the literature on fermentation or strain due to limited information on methods of obtaining extracts obtained from cherry fruits. The effect of the fermentation process should be supported by trials under controlled conditions.