Türkiye’ye Özgü Yöresel Bir Pancar Çeşidinde Ve Atıklarında Fenolik Madde Profilinin, Antioksidan Kapasitesinin Ve Biyoyararlılığın Belirlenmesi
Türkiye’ye Özgü Yöresel Bir Pancar Çeşidinde Ve Atıklarında Fenolik Madde Profilinin, Antioksidan Kapasitesinin Ve Biyoyararlılığın Belirlenmesi
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Tarih
2015-02-25
Yazarlar
Kasapoğlu, Kadriye Nur
Süreli Yayın başlığı
Süreli Yayın ISSN
Cilt Başlığı
Yayınevi
Fen Bilimleri Enstitüsü
Institute of Science and Technology
Institute of Science and Technology
Özet
Sebze tüketimi, sebzelerin sahip olduğu sayısız besleyici içerik sayesinde sağlıklı ve geleneksel bir beslenme biçimidir. Sebzelerin ve meyvelerin bu sağlığa yararlı etkisi fitokimyasallar, vitaminler ve enzimlerin yanı sıra mineraller ve life de dayanır. Bu bileşimler kanser ve kardiyovasküler hastalık risklerine karşı içerdikleri zengin antioksidanlar sayesinde yüksek koruma sağlarlar. Polifenoller meyve ve sebzenin içinde diğer bileşenlere ek olarak antioksidan aktivite sağlarlar. Antioksidan potansiyel gösteren polifenolleri taşıyan bitkiler giderek araştırmacıların daha çok dikkatini çekmektedir. Kırmızı pancar (Beta vulgaris L.) fenoliklerin yanı sıra suda-çözünür azot ihtiva eden antioksidan, anti-mikrobiyal, anti-viral, anti-prolifetarif, anti-iltihabik, anti-kanser ve karaciğeri koruyucu etkilere sahip olduğu bilinen betalainler olarak adlandırılan pigmentleri içermektedir. Tüm bu sağlığa olumlu etkileri dolayısıyla, yeni betalain içeren gıda ürünlerine karşı artan ilgi söz konusudur. Türkiye'ye özgü bir yöresel kırmızı pancar çeşidinde iç kısım (BVf) ve kabuklar (BVp) bu çalışma kapsamında ilk kez araştırılmıştır. Bilgimize göre, bu varyeteye ait herhangi bir çalışma literatürde bulunmamaktadır. Deneyler, farklı zamanlarda ayrı yerlerden hasat edilen numunelerle üç farklı biyolojik tekrar olarak yapılmıştır. Tekrar yığınların sonuçları arasında ve kabuk ile iç kısım arasında tüm deneylere göre istatistiksel olarak önemli farklar izlenmiştir. Kabuk kısımları bütün analizlerde (toplam fenolik, toplam betalain, ve antioksidan kapasite) iç kısımlardan daha yüksek değerler vermiştir. Fenolik profil UFLC-MS/MS kullanılarak analiz edilmiştir. Kabuk ve iç kısımlarda benzer fenolik asit kromatogramları gözlenmiş olup, fenolik asit miktarları yığınlara göre farklılık göstermiştir. Sonuç olarak, sinnamik asit, klorojenik asit, vanilik asit, 4-hidroksi-benzoik asit, trans-ferulik asit ve kafeik asit hem iç hem kabuk kısımlarında farklı konstrasyonlarda izlenmiştir. Bununla birlikte, kafeik asit ve rutin sadece iz miktarda gözlenmiştir. Özellikle, klorojenik ve ferulik asit kabuk kısımları için en yüksek miktardaki fenolik bileşenlerdir. Kırmızı pancarda ise en yüksek miktardaki fenolik asit 4(p-)hidroksi-benzoik asit olarak bildirilmiştir. Bu varyetenin fenolik asit kompozisyonunun kırmızı pancar ve şeker pancarından farklı olduğu bulunmuştur. Kabuk için fenolik asit miktarları azalan sırayla: ferulik asit, klorojenik asit, şirincik asit, 4(p-)hidroksibenzoik asit, vanilik asit olarak izlenirken, iç kısım içinse: ferulik asit, şirincik asit, , vanilik asit, 4(p-)hidroksibenzoik asit, ve klorojenik asit olarak izlenmiştir. Gallik asit, elajik asit, prokateşuik asit, sinapik asit, trans-sinamik asit, ve p-kumarik asit ise tespit edilememiştir. Toplam antioksidan kapasite analizlerinde, literatürle benzer şeklide DPPH yöntemi, CUPRAC yönteminden daha düşük sonuçlar vermiştir. Ancak, antioksidan kapasitenin kırmızı pancarda olduğu kadar yüksek olmadığı görülmüştür. Toplam fenolik madde miktarı kabuk için 8.50 mg GAE/ g DW iken iç kısım için 5.45 mg GAE/ g DW olarak bulunmuştur. Bu değerler yüksek olmamakla birlikte, kırmızı pancar için literatürde bildirilen aralığa girmektedir. Spektrofotometrik yöntemler olan DPPH ve CUPRAC metodları toplam antioksidan kapasitesini belirlemek için kullanılmıştır. Literatürde antioksidan kapasite tayini için pancar numunelerinde en çok DPPH yöntemi kullanılmıştır. CUPRAC metodu ise asetat tampon çözelti sayesinde fizyolojik pH ortamında analiz sağladığı için sebze-meyve numunelerinde tercih edilmektedir. Toplam fenolik madde içeriği ise Folin-Ciocalteu metodu ile belirlenmiştir. Folin-Ciocalteu metodu, numunede bulunan fenolikleri gallik asit eşdeğeri cinsinden toplam miktar olarak tayin eder. Pancarın hem iç kısım hem de kabuklarında bulunan fenoliklerin biyoyararlılığı in vitro sindirim metodu kullanılarak ağız, mide ve ince bağırsak sindirimi fizyokimyasal ve biyokimyasal anatomi simüle edilerek incelenmiştir. Insan ve hayvan gibi canlı organizmada yapılan araştırmalar karmaşıklık, pahalılık ve etik sebeplerden dolayı tercih edilmediğinden, in vitro metodu deney ortamında sindirimi çözümleyip hızlı ve basit yöntem sunarak bitki matrislerinde değerlendirmede olanağı sağlar. In vitro sindirim metodu sonuçlarına göre, yığınların biyoyararlılığı arasındaki farklar önemlidir. Ayrıca, kabuk numunelerde fenolik madde miktarındaki azalma iç kısımlardan fazladır. Yani, iç kısımların biyoyararlılığı kabuk kısımlarından yüksektir. Fenolik asitler mide sindiriminde asidik ortam şartlarından dolayı stabildirler. Ancak, IN fraksiyonlarında, yani bağırsak içinde emilmek üzere bulunan fenolik miktarlarında azalma meydana gelmekle beraber, incelenen numunede bulunan fenolikler kullanılabilir orandadırlar. In vitro sindirim sonrası UFLC-MS/MS analizinde 4(p-)hidroksibenzoik asit IN, PG ve OUT fraksiyonlarında izlenmemiştir. Toplam betalain içeriği spektrofotometrik yöntem ile ölçülerek kırmızı-mor (betacyanin -BC) ve sarı-turuncu (betaxanthin -BX) pigmentlerin toplamı olarak ifade edilmiştir. Betaxantin ve betasiyanin içeriği kabuk için sırasıyla 1.6 ± 0.01 mg g-1 indicaxantin eşdeğeri ve 0.9 ± 0.01 mg g-1 betanin eşdeğeri olarak bulunurken, iç kısım için 0.3 ± 0.05 mg g-1 indicaxantin ve 0.9 ± 0.01 mg g-1 betanin olarak bulunmuştur. Toplam betalain içeriği kabuk için 2.5 mg/ g ve iç kısım için 1.2 mg/ g olarak bulunmuştur. Bu sonuç, kırmızı pancara göre oldukça düşük olmakla beraber, literatürde araştırılan diğer betalain içeren bitki, sebze ve meyvelere kıyasla değerlidir. Güncel olarak, sağlık bakımından pek çok fayda ihtiva eden betalain pigmenti eldesiyle ilgili pek çok çalışma yapılarak yeni kaynaklar araştırılmaktadır. Bu bağlamda, yalnızca toplam betalain içeriği değil, BX/BC pigmentlerinin oranı da farklı renk skalası elde etme bakımından önem teşkil etmektedir.
Consumption of vegetables is considered as a traditional healthy diet because of the numerous nutrient substances they possessed. The health promoting action of fruits and vegetables has been attributed to the presence of phytochemicals, vitamins and enzymes, as well as other minerals and fibers. These compounds have strong protective effects against major disease risks including cancer and cardiovascular diseases due to their rich potential as antioxidants. Polyphenols are present in fruits and vegetables in great abundance as powerful antioxidants in addition to the other constituents. Plants bearing polyphenols and other phytochemicals with antioxidant potential are gaining attention of researchers gradually. Beetroot (Beta vulgaris L.) contains water-soluble nitrogen-containing pigments called as betalains as well as polyphenolics compounds. Betalains are known to have antioxidant, antimicrobial, antiviral, antiproliferative, anti-inflammatory, hepatoprotective, and anti-cancer activities. Due to these excessive health-promoting effects, new betalain-containing food products are of interest. A local red beet variety native to Turkey has been investigated for the first time including fleshes (BVf) and peels (BVp). The experiments were carried out to three different biological replicates which were harvested at different times from various farms. The results of all assays indicated statistically significant differences between the batches and between peel and flesh samples. Peel samples showed higher values than flesh samples according to all assays performed. Phenolic profile was analyzed using UFLC-MS/MS. Similar chromatogram patterns of phenolic acids were observed for peel and flesh samples; contents differed depending on batches. As a result, the presence of chlorogenic acid, ferulic acid, syringic acid, vanillic acid and 4(p-)hydroxybenzoic acid were detected in both types of extract (BVp and BVf) at different concentrations. However, caffeic acid and rutin were present only in trace amounts. Notably, chlorogenic and ferulic acid was the major phenolic constituents for BVp samples whereas ferulic acid was the only major phenolic in BVf samples in contrast with red beetroot, which contains 4-hydroxybenzoic acid as the major phenolic acid. The phenolic acid composition was seen different from the red beet and sugar beet. The decreasing order of individual phenolic acids in peels was as follows: ferulic acid, chlorogenic acid, syringic acid, 4(p-)hydroxybenzoic acid, vanilic acid. On the other hand, the decreasing order of individual phenolic acids in fleshes was as Ferulic acid, syringic acid, vanilic acid, 4(p-)hydroxybenzoic acid, chlorogenic acid. In regards to the total antioxidant capacity methods, CUPRAC assay gave higher responses than the DPPH assay in accordance with the literature, although the antioxidant capacity of this beetroot variety was not as high as red beetroot. Total phenolic content was in a similar range with the reported values for red beetroot. DPPH Free Radical Scavenging Capacity and CUPRAC Cupric Reducing Antioxidant Capacity assays were performed to determine the antioxidant capacity and the total phenolic content was determined via Folin-Ciocalteu method. The bioaccessibility of phenolics from both flesh and peel parts was assessed by in vitro gastro-intestinal digestion procedure that simulates the physiochemical and biochemical changes that occur in the gastrointestinal tract. Since in vivo studies on humans and animals are not preferred due to the limitations such as complexity, expensiveness and ethical reasons, in vitro digestion assays provide rapid and simple methods to evaluate phytochemical stability in food matrices. Results from in vitro digestion demonstrated that differences in bioaccessibility of polyphenolics between the batches were significant and the reduction of TPC in peel samples is greater than in fleshes. In other words, the bioaccessibility of phenolics in fleshes is greater than the bioaccessibility of phenolics in fleshes. Additionally, phenolic acids were found to be stable in gastric digestion due to acidic conditions. However, their recoveries become lowered in IN fractions representing the compound passed into serum. The UFLC-MS/MS detection was done on phenolic acids from PG, IN and OUT fractions of GI digestion. Notably, no peak could be detected in IN, OUT and PG fractions for the 4(p-)hydroxybenzoic acid. Furthermore, the total betalain content was quantified spectrophotometrically and expressed as the sum of red-violet (betacyanin -BC) and yellow-orange (betaxanthin -BX) pigments. BX and BC of peel samples were found as 1.6 ± 0.01 mg g-1 indicaxanthin equivalent and 0.9 ± 0.01 mg g-1 betanin equivalent whereas 0.3 ± 0.05 mg g-1 indicaxanthin equivalent and 0.9 ± 0.01 mg g-1 betanin equivalent for flesh samples, respectively.Total betalain content of this beetroot variety was found as 2.5 mg/ g for peels and 1.2 mg/ g for fleshes, lower than the red beetroot.
Consumption of vegetables is considered as a traditional healthy diet because of the numerous nutrient substances they possessed. The health promoting action of fruits and vegetables has been attributed to the presence of phytochemicals, vitamins and enzymes, as well as other minerals and fibers. These compounds have strong protective effects against major disease risks including cancer and cardiovascular diseases due to their rich potential as antioxidants. Polyphenols are present in fruits and vegetables in great abundance as powerful antioxidants in addition to the other constituents. Plants bearing polyphenols and other phytochemicals with antioxidant potential are gaining attention of researchers gradually. Beetroot (Beta vulgaris L.) contains water-soluble nitrogen-containing pigments called as betalains as well as polyphenolics compounds. Betalains are known to have antioxidant, antimicrobial, antiviral, antiproliferative, anti-inflammatory, hepatoprotective, and anti-cancer activities. Due to these excessive health-promoting effects, new betalain-containing food products are of interest. A local red beet variety native to Turkey has been investigated for the first time including fleshes (BVf) and peels (BVp). The experiments were carried out to three different biological replicates which were harvested at different times from various farms. The results of all assays indicated statistically significant differences between the batches and between peel and flesh samples. Peel samples showed higher values than flesh samples according to all assays performed. Phenolic profile was analyzed using UFLC-MS/MS. Similar chromatogram patterns of phenolic acids were observed for peel and flesh samples; contents differed depending on batches. As a result, the presence of chlorogenic acid, ferulic acid, syringic acid, vanillic acid and 4(p-)hydroxybenzoic acid were detected in both types of extract (BVp and BVf) at different concentrations. However, caffeic acid and rutin were present only in trace amounts. Notably, chlorogenic and ferulic acid was the major phenolic constituents for BVp samples whereas ferulic acid was the only major phenolic in BVf samples in contrast with red beetroot, which contains 4-hydroxybenzoic acid as the major phenolic acid. The phenolic acid composition was seen different from the red beet and sugar beet. The decreasing order of individual phenolic acids in peels was as follows: ferulic acid, chlorogenic acid, syringic acid, 4(p-)hydroxybenzoic acid, vanilic acid. On the other hand, the decreasing order of individual phenolic acids in fleshes was as Ferulic acid, syringic acid, vanilic acid, 4(p-)hydroxybenzoic acid, chlorogenic acid. In regards to the total antioxidant capacity methods, CUPRAC assay gave higher responses than the DPPH assay in accordance with the literature, although the antioxidant capacity of this beetroot variety was not as high as red beetroot. Total phenolic content was in a similar range with the reported values for red beetroot. DPPH Free Radical Scavenging Capacity and CUPRAC Cupric Reducing Antioxidant Capacity assays were performed to determine the antioxidant capacity and the total phenolic content was determined via Folin-Ciocalteu method. The bioaccessibility of phenolics from both flesh and peel parts was assessed by in vitro gastro-intestinal digestion procedure that simulates the physiochemical and biochemical changes that occur in the gastrointestinal tract. Since in vivo studies on humans and animals are not preferred due to the limitations such as complexity, expensiveness and ethical reasons, in vitro digestion assays provide rapid and simple methods to evaluate phytochemical stability in food matrices. Results from in vitro digestion demonstrated that differences in bioaccessibility of polyphenolics between the batches were significant and the reduction of TPC in peel samples is greater than in fleshes. In other words, the bioaccessibility of phenolics in fleshes is greater than the bioaccessibility of phenolics in fleshes. Additionally, phenolic acids were found to be stable in gastric digestion due to acidic conditions. However, their recoveries become lowered in IN fractions representing the compound passed into serum. The UFLC-MS/MS detection was done on phenolic acids from PG, IN and OUT fractions of GI digestion. Notably, no peak could be detected in IN, OUT and PG fractions for the 4(p-)hydroxybenzoic acid. Furthermore, the total betalain content was quantified spectrophotometrically and expressed as the sum of red-violet (betacyanin -BC) and yellow-orange (betaxanthin -BX) pigments. BX and BC of peel samples were found as 1.6 ± 0.01 mg g-1 indicaxanthin equivalent and 0.9 ± 0.01 mg g-1 betanin equivalent whereas 0.3 ± 0.05 mg g-1 indicaxanthin equivalent and 0.9 ± 0.01 mg g-1 betanin equivalent for flesh samples, respectively.Total betalain content of this beetroot variety was found as 2.5 mg/ g for peels and 1.2 mg/ g for fleshes, lower than the red beetroot.
Açıklama
Tez (Yüksek Lisans) -- İstanbul Teknik Üniversitesi, Fen Bilimleri Enstitüsü, 2015
Thesis (M.Sc.) -- İstanbul Technical University, Instıtute of Science and Technology, 2015
Thesis (M.Sc.) -- İstanbul Technical University, Instıtute of Science and Technology, 2015
Anahtar kelimeler
Fenolik Asit,
Fenolik Profil,
Betalain,
Antioksidan Aktivite,
Biyoyararlılık,
Phenolic Acid,
Phenolic Profile,
Betalain,
Antioxidant Capacity,
Bioaccessibility