Malaysian Journal of Analytical Sciences Vol 22 No 5 (2018): 785 - 793

SEPARATION OF L-ARGININE AND L-CITRULLINE IN RED AND YELLOW CRIMSON WATERMELON (Citrullus lanatus) JUICES EXTRACT USING HPLC GRADIENT MODE

(Pemisahan L-Arginina dan L-Sitrulina dalam Ekstrak Jus Tembikai (Citrullus lanatus) Merah dan Kuning Menggunakan Mod Cerun KCPT)

Rasdin Ridwan¹, Hairil Rashmizal Abdul Razak², Mohd Ilham Adenan^3,4, Wan Mazlina Md Saad¹*

¹Centre of Medical Laboratory Technology, Faculty of Health Sciences,

Universiti Teknologi MARA Selangor, Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor, Malaysia

²Centre for Diagnostic Nuclear Imaging, Faculty of Medicine & Health Sciences,

Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

³Faculty of Applied Sciences,

Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia

⁴Atta-ur-Rahman Institute for Natural Product Discovery, Aras 9, Bangunan FF3,

Universiti Teknologi MARA Selangor, Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor, Malaysia

*Corresponding author: wanmaz755@salam.edu.uitm.my

Received: 29 August 2017; Accepted: 28 August 2018

Abstract

Watermelon (Citrullus lanatus) is a nutritious fruit that has attracted scientific interest due to its phytonutrient content including carotenoids and amino acids. Watermelon juice is the richest known source of amino acids including L-arginine and L-citrulline. For a complete separation of amino acids in watermelon juices, a gradient mode of reverse phase high performance liquid chromatography (RP-HPLC) with UV detection was utilised owing to its supremacy over isocratic mode. This present study was aimed at achieving good separation of chromatography profiles and baseline between L-arginine and L-citrulline in red and yellow crimson watermelon juice extract. The edible parts of watermelons were juiced, stored at – 80 ^oC and freeze dried to obtain dried juice powder. Samples were analysed by gradient mode of RP-HPLC using 0.1% orthophosphoric acid in water (v/v) and acetonitrile as mobile phase A and B. The chromatograph separation was performed using Gemini C₁₈ column at a flow rate of 1.0 mL/min, maintained temperature at 25 ^oC while detection was made at four different wavelengths; 195 nm, 200 nm, 207 nm and 210 nm with total analysis time of 35 min. Excellent chromatographic separation was achieved at conditions of gradient elution (0%B - 95%B, 0-18 minutes and 95%B, 18-23 minutes, 95%B-0%B, 23-25 minutes, 0%B, 25-35 minutes) with baseline between L-arginine and L-citrulline in both watermelon juice extracts. The gradient mode used in this RP-HPLC successfully achieved efficient separation of chromatographic profile for amino acids, L-arginine and L-citrulline in the red and yellow crimson watermelon juice extracts.

Keywords: separation, L-arginine, L-citrulline, watermelon, juices extract

Abstrak

Tembikai (Citrullus lanatus) adalah buah yang berkhasiat yang menarik perhatian saintifik kerana kandungan fitonutriennya seperti karotenoid dan asid amino. Jus tembikai mempunyai asid amino yang tinggi termasuk L-arginina dan L-sitrulina. Untuk pemisahan lengkap asid amino dalam ekstrak jus tembikai, mod cerun fasa terbalik kromatografi cecair prestasi tinggi (FT-KCPT) dengan pengesanan UV dijalankan kerana kehebatannya berbanding mod isokratik. Kajian ini bertujuan mencapai pemisahan yang baik bagi profil kromatografi dan garis dasar antara L-arginina dan L-sitrulina dalam ekstrak jus tembikai merah dan tembikai kuning. Isi buah tembikai ditapis, disimpan di – 80 ^oC dan diletakkan dalam proses pengeringan-beku untuk mendapatkan serbuk jus kering. Sampel dianalisis menggunakan mod cerun FT-KCPT dengan 0.1% asid ortofosforik dicairkan dalam air (v/v) dan asetonitril sebagai fasa bergerak A dan B. Pemisahan kromatografi dilakukan dengan menggunakan turus Gemini C₁₈ pada kadar aliran 1.0 mL/min, suhu yang dikekalkan pada 25 ^oC dan pengesanan dibuat pada empat panjang gelombang yang berbeza; 195 nm, 200 nm, 207 nm dan 210 nm dengan 35 minit masa analisis keseluruhan. Pemisahan kromatografi yang cemerlang dicapai dengan keadaan elusi kecerunan (0% B - 95% B, 0-18 minit dan 95% B, 18-23 minit, 95% B-0% B, 23-25 minit, 0% B, 25-35 minit) dengan garis dasar antara L-arginina dan L-sitrulina dalam kedua-dua ekstrak jus tembikai. Mod cerun dalam FT-KCPT ini berjaya mencapai pemisahan profil kromatografi yang efisien untuk asid amino, L-arginina dan L-sitrulina dalam ekstrak jus tembikai merah dan kuning.

Kata kunci: pemisahan, L-arginina, L-sitrulina, tembikai, ekstrak jus

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