İçeceklerde Askorbik ve Oksalik Asitlerin Eşzamanlı Analizi İçin HPLC-UV Yönteminin Validasyonu

Year 2024, Volume: 22 Issue: 4, 273 - 281, 31.12.2024

Gülşen Aksin Biltekin Abdullah Akdoğan , Mustafa Soylak , Ümit Divrikli

https://doi.org/10.24323/akademik-gida.1609556

Abstract

Ters faz yüksek performanslı sıvı kromatografisi (RP/HPLC-UV) yöntemi, endüstriyel ve taze sıkılmış içeceklerde askorbik ve oksalik asitlerin hızlı ve eş zamanlı analizi için geliştirilmiştir. Askorbik asidin hızlı bozulması nedeniyle, stabilitesi de araştırılmıştır. Analiz, pH 2.3’te 0.01 mol L⁻¹ KH₂PO₄ fosfat tamponundan oluşan hareketli faz ile Inertsil ODS-3 kolon kullanılarak gerçekleştirilmiştir. Tespit dalga boyları askorbik asit için 245 nm ve oksalik asit için 205 nm olarak ayarlanmıştır. Tespit limitleri (LOD), askorbik asit için 1.4 mg L⁻¹ ve oksalik asit için 1.3 mg L⁻¹ olarak belirlenmiştir. Endüstriyel meyve suyu paketinin açılmasından 48 saat sonra, askorbik asit içeriğinde %18.5’lik bir azalma tespit edilmiştir. Yöntem, gün içi tekrarlanabilirlik (n= 3) açısından askorbik asit için %4.1 ve oksalik asit için %4.6 oranında bir doğruluk göstermiştir. Günler arası hassasiyet, bağıl standart sapma (RSD) olarak ifade edilmiş ve 10 tekrar (n= 10) temelinde askorbik asit için %3.85 ve oksalik asit için %4.33 olarak hesaplanmıştır. Yöntemin doğruluğu, %93 ile %104 arasında değişen ortalama geri kazanım oranı ile doğrulanmıştır.

Keywords

Sıvı kromatografisi, Askorbik asit, Oksalik asit, İçecek, Meyve suyu

Supporting Institution

Pamukkale University

Project Number

2023FEBE020

Thanks

The authors would like to thank the Pamukkale University for financial support to this research project (Project no 2023FEBE020).

Validation of an HPLC-UV Method for Simultaneous Analysis of Ascorbic and Oxalic Acids in Beverages

Abstract

A reversed-phase high-performance liquid chromatography (RP/HPLC-UV) method was developed for the rapid and simultaneous analysis of ascorbic and oxalic acids in both industrial and freshly squeezed beverages. Due to the rapid degradation of ascorbic acid, its stability was also investigated. The analysis was performed using an Inertsil ODS-3 column with a mobile phase consisting of 0.01 mol L⁻¹ KH₂PO₄ phosphate buffer at pH 2.3. Detection wavelengths were set at 245 nm for ascorbic acid and 205 nm for oxalic acid. The limits of detection (LOD) were 1.4 mg L⁻¹ for ascorbic acid and 1.3 mg L⁻¹ for oxalic acid. After 48 hours of opening an industrial fruit juice package, an 18.5% reduction in ascorbic acid content was found. The method demonstrated within-day repeatability (n = 3) of 4.1% for ascorbic acid and 4.6% for oxalic acid. Between-day precision, expressed as relative standard deviation (RSD), was 3.85% for ascorbic acid and 4.33% for oxalic acid, based on 10 replicates (n= 10). The accuracy of the method was validated with an average recovery rate ranging from 93% to 104%.

Keywords

Liquid chromatography, Ascorbic acid, Oxalic acid, Beverage, Fruit juice

Project Number

2023FEBE020

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