Developing and validation of an HPLC-DAD method for the determination of eight phenolic constituents in extract of different wine species

Abstract

ÖZAmaç: Bu çalışmada amaç bazı yerel şarap ve meyve şarabı örneklerinde gallik asit, klorojenik asit, epigallokateşin, kafeik asit, vanilin, p-kumarik asit, rutin ve kersetin fenolik bileşiklerinin tayini için yeni bir HPLC yöntemi geliştirilmesi ve geçerli kılınmasıdır. Gereç ve Yöntemler: Analizler, gradyan sistem kullanılarak bir Zorbax Eclipse C18 kolon (4.6 x 150 mm, 3.5-μm partikül boyutlu) üzerinde yürütülmüştür. Akış hızının 1 mL/min olduğu sistemde hareketli faz A 10-mM fosforik asit ve hareketli faz B metanol olarak belirlenmiştir. Fenolik bileşikler, üç farklı dalga boyunda bir DAD kullanılarak izlenmiştir. Bulgular: Geliştirilen ve geçerli kılınan yöntemin 1-100 ppm derişim aralığında doğrusal cevap verdiği gözlenmiştir. Geri kazanım değerleri %95-105 değerleri arasındadır ve tekrarlanabilir sonuçlardır. Geliştirilen yöntem çeşitli şarap örneklerinde fenolik profili incelemek için uygulanmıştır. Sonuç: Yapılan çalışma sonucunda doğru, hassas ve güvenilir ölçüm yapan bir HPLC-DAD yöntemi geliştirilmiş ve şarap özütlerinde, şarabın antioksidan özellik göstermesini sağlayan fenolik maddelerin derişimlerini tayin etmek için başarıyla kullanılmıştır. Anahtar kelimeler: Fenolik bileşikler, tayin, şarap, HPLC, validasyonObjectives: A new HPLC method was developed and validated for the determination of some phenolic compounds; gallic acid, chlorogenic acid, epigallocatechin, caffeic acid, vanillin, p-coumaric acid, rutin, and quercetin in some local wine and fruit wine samples. Materials and Methods: Analyses were performed on a Zorbax Eclipse C18 column (4.6 x 150 mm, 3.5-μm particle size) using a gradient system. Mobile phase A was a 10-mM phosphoric acid solution and mobile phase B was methanol using a flow rate of 1 mL/min. Phenolic components were monitored using a DAD at three different wavelengths. Results: The developed and validated method was generally linear between the 1-100 ppm concentration range. Recovery values were obtained in the range of 95-105% and repetitive. The method was successfully applied to investigate the phenolic profiles of different wine samples. Conclusion: As a result of the study, an accurate, sensitive and reliable HPLC-DAD method was developed. The method was successfully used to determine the concentrations of antioxidant phenolic constituents from some local wine extracts.

Development of a Multifunction Set Yogurt Using Rubus suavissimus S. Lee (Chinese Sweet Tea) Extract

“…The identification and quantification of the individual phenolic compounds were carried out using an Agilent-1260 HPLC (USA) equipped with a C18 column (Kinetex ® 5 µm EVO 100 × 4.6 mm, Phenomenex, Torrance, CA, USA) [ 19 ]. In brief, 5 g of Chinese sweet tea extract was dissolved in 50 mL of methanol and centrifuged at 6000× g for 10 min.…”

Development of HPLC Method for Catechins and Related Compounds Determination and Standardization in Miang (Traditional Lanna Fermented Tea Leaf in Northern Thailand)

“…Column B (deactivated, endcapped monomeric C18, high-purity silica) was tested using methanol in the presence and absence of ethyl acetate mixed with either phosphoric acid or acetic acid as mobile phases modified from a previous report [ 33 ]. The separation was performed with ethyl acetate added to the mobile phase gave sharper peak shapes (C, EGCG, and GCG) than those with ethyl acetate absent [ 34 , 35 , 36 ].…”

A Concise Profile of Gallic Acid—From Its Natural Sources through Biological Properties and Chemical Methods of Determination

“…Currently, many different techniques are used to isolate GA. An overview of the representative GA isolation conditions is presented in Table 3 . These include classical approaches, such as distillation [ 109 ], maceration [ 88 , 89 , 90 , 91 , 92 , 94 , 97 , 100 , 101 , 103 , 106 , 108 , 109 , 112 , 116 , 118 , 120 , 123 ], reflux extraction [ 27 , 93 , 103 , 106 , 107 , 114 ], reactive extraction [ 102 ], enzyme-assisted or polyamide membranes extraction [ 101 ] as well as newer and more advanced ones, such as ultrasound-assisted extraction [ 98 , 99 , 115 , 122 ], microwave-assisted extraction [ 103 , 111 ], and simpler and miniaturized techniques such as LLME [ 118 ], SPE [ 114 ], or MSPD [ 113 ].…”

“…Determination of gallic acid by column chromatography is most often performed using high-performance liquid chromatography (HPLC) [ 26 , 29 , 64 , 83 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 ], although its more advanced form, i.e., ultra-performance liquid chromatography (UPLC), is also used [ 28 ]. These techniques allow the determination of very low analyte concentrations in the presence of sometimes many co-eluting components, often with the properties very similar to these of GA. Their great advantage in the analysis of compounds in plant and biological matrices is the wide range of commercially available columns, which allows one to control the selectivity of the separation process depending on the needs of a given analytical challenge and, owing to the high separation efficiency, the use of smaller amounts of solvents.…”

“…These techniques allow the determination of very low analyte concentrations in the presence of sometimes many co-eluting components, often with the properties very similar to these of GA. Their great advantage in the analysis of compounds in plant and biological matrices is the wide range of commercially available columns, which allows one to control the selectivity of the separation process depending on the needs of a given analytical challenge and, owing to the high separation efficiency, the use of smaller amounts of solvents. It should also be remembered that liquid column chromatography does not usually require derivatization of the substances to be determined because, due to the natural ability of most biologically active compounds of natural origin to absorb electromagnetic radiation, separations are most often monitored using a UV [ 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 103 ] or DAD [ 88 , 99 , 102 ] detector. Considering all the above unique properties, HPLC is recognized as one of the standard methods for the analysis of many compounds, including gallic acid analysis.…”