Malaysian Journal of Analytical Sciences Vol 26 No 2 (2022): 360 - 369

 

 

 

 

DEVELOPMENT AND OPTIMIZATION OF A RAPID RESOLUTION LIQUID CHROMATOGRAPHY METHOD FOR CYANIDIN-3-O-GLUCOSIDE IN RAT PLASMA

 

(Pembangunan dan Pengoptimuman Kaedah Kromatografi Cecair Resolusi Pantas untuk Sianidin-3-O-Glukosida Klorida di dalam Plasma Tikus)

 

Nadiratul Asyikin Sauji¹, Wan Amir Nizam Wan Ahmad¹, Liza Nordin², Ruzilawati Abu Bakar³*

 

¹Biomedicine Program, School of Health Sciences

²Department of Physiology, School of Medical Sciences

³Department of Pharmacology, School of Medical Sciences

Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

 

*Corresponding authors:  ruzila@usm.my

 

 

Received: 16 January 2022; Accepted: 15 March 2022; Published:  28 April 2022

 

 

Abstract

The growing interest in anthocyanins in plants has brought about the importance of investigating their pharmacological properties. Sensitive and specific analytical methods are required to accurately analyze the anthocyanins present in samples. One of the anthocyanins found in plants is cyanidin-3-O-glucoside. The objective of this study was to develop and optimize a rapid resolution liquid chromatography (RRLC) method for cyanidin-3-O-glucoside determination in rat plasma. Spectrophotometric analysis was performed to determine the best ultraviolet (UV) absorbance wavelength. Liquid-liquid extraction (LLE) and solid-phase extraction (SPE) methods were compared to determine the best extraction method for cyanidin-3-O-glucoside in rat plasma samples. The effects of varying the type and proportion of organic solvents, the type and concentration of buffer solutions, flow rates, column temperatures, and UV wavelengths were examined. The optimized chromatographic method for RRLC analysis of cyanidin-3-O-glucoside was a mobile phase composition of 0.1% trifluoroacetic acid aqueous solution and acetonitrile in a ratio of 81:19, respectively, with a 0.5 mL/min flow rate, at 30°C column temperature and 525 nm detection wavelength. SPE was our choice of final extraction method. Our findings revealed that the optimized RRLC method can be used to determine cyanidin-3-O-glucoside in rat plasma.

 

Keywords:  rapid resolution liquid chromatography, method development, cyanidin-3-o-glucoside, rat plasma

 

Abstrak

Kajian antosianin dalam rosel yang semakin meluas mengetengahkan kepentingan analisis sebatian tersebut untuk mengkaji ciri-ciri farmakologinya. Kaedah analisis yang sensitif dan spesifik diperlukan untuk menganalisis antosianin yang terdapat dalam sampel dengan tepat. Salah satu antosianin yang terdapat dalam tumbuhan ialah sianidin-3-O-glukosida. Objektif kajian ini adalah untuk membangunkan dan mengoptimumkan kaedah kromatografi cecair resolusi pantas (RRLC) untuk sianidin-3-O-glukosida di dalam plasma tikus. Analisis spektrofotometri dilakukan untuk memilih penyerapan ultraungu yang terbaik. Kaedah pengekstrakan cecair-cecair (LLE) dan pengekstrakan fasa pepejal (SPE) juga dijalankan untuk menilai kaedah pengekstrakan terbaik bagi antosianin daripada sampel plasma tikus. Kesan mempelbagaikan jenis dan peratusan pelarut organik, jenis dan kepekatan larutan penimbal, kadar aliran fasa bergerak, suhu turus dan panjang gelombang pengesan ultraungu telah diuji. Kaedah pengoptimuman kromatografi menunjukkan komposisi fasa bergerak bagi larutan akueus asid trifluoroasettik 0.1% dan asetonitril dalam nisbah 81:19, dengan kadar aliran 0.5 mL/min, pada suhu turus 30°C dan panjang gelombang pengesanan 525 nm adalah sesuai untuk analisis sianidin-3-O-glukosida. Kaedah SPE dipilih sebagai kaedah pengekstrakan terbaik kerana ia menghasilkan puncak kromatogram yang lebih baik berbanding kaedah LLE. Kesimpulannya, kaedah RRLC yang dibangunkan dalam kajian ini boleh digunakan untuk menentukan sianidin-3-O-glukosida dalam plasma tikus.

 

Kata kunci:  kromatografi cecair resolusi pantas, pembangunan kaedah, sianidin-3-O-glukosida, plasma tikus

 

 

 

Graphical Abstract

 

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