Ekrem Cem Çankırılıgil, NERMİN BERİK, Esen Alp ERBAY

Optimization of hydrolyzation procedure for amino acid analysis in fish meat with HPLC-DAD by Response Surface Methodology (RSM)

Abstract: In this research, with the aim of maximizing amino acid content by different hydrolyzation procedures of fish meat were carried out and analysed with high performance liquid chromatography (HPLC) equipped with diode array detector (DAD). Response surface methodology (RSM) was used to determine factors that used in the experiments. The RSM suggested 16 different hydrolyzation trials between the ranges of normality as 3 N - 8 N, temperature as 90°C110°C and duration as 12-24 hours were run. In the following, hydrolysed samples were analysed with HPLC-DAD and obtained data were evaluated with Box-Behnken method in RSM. Our results show that, the most effective experiment was found as hydrolysing by 3.79 N at 110°C in 24 hours for total amino acid content as well as maximized essential amino acids with 0.981 desirability. For sensitive ones, hydrolysing in 3.42 N at 106.8°C in 12.02 hours was found as most effective with maximized Asn, Gln and Trp with 0.849 desirability.

Balık etinde HPLC-DAD ile yapılan amino asit analizi hidrolizasyon prosedürünün Yanıt Yüzey Metodu (RSM) ile optimizasyonu

Bu çalışmada; diyot dedektörü (DAD) donanımlı yüksek performanslı likit kromatografi (HPLC) ile balık etinde gerçekleştirilen amino asit analizinde, amino asit miktarını maksimize etmek için farklı hidrolizasyon prosedürleri gerçekleştirilmiştir. Deneylerde kullanılan faktörleri belirlemek için yanıt yüzey metodu (RSM) kullanılmıştır 16 adet farklı hidrolizasyon deneyi RSM ile kurulmuş olup, 3N-8N arasındaki normalite değerleri, 90°C-110°C arasındaki sıcaklık değerleri ve 12-24 saat arasındaki süre değerleri dikkate alınmıştır. Elde edilen hidrolizatlar HPLC-DAD ile analiz edilmiş olup elde edilen veri RSM’deki Box-Behnken metoduna göre değerlendirilmiştir. Sonuçlara göre; toplam ve esansiyel amino asitler için en efektif hidrolizasyon metodu 3,29 normalitede, 110°C sıcaklıkta, 24 saat olarak belirlenmiş olup istenilebilirlik değeri 0,981 olarak tespit edilmiştir. Hassas amino asitler için ise; 3,42 normalitede, 106,8°C sıcaklıkta, 12,02 saat süre Asn, Gln ve Trp için maksimum değerleri vermiş olup istenilebilirlik değeri 0,849 olarak tespit edilmiştir.

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