Sains Malaysiana 44(1)(2015): 115–120

A Simple and Effective Isocratic HPLC Method for Fast Identification and

Quantification of Surfactin

(Kaedah Isokratik HPLC Ringkas dan Berkesan bagi Pengenalpastian dan Kuantifikasi Surfaktin dengan Cepat)

MUHAMMAD QADRI EFFENDY MUBARAK¹, ABDUL RAHMAN HASSAN¹, AIDIL ABDUL HAMID², SAHAID KHALIL³ & MOHD HAFEZ MOHD ISA¹*

¹Faculty of Science and Technology, Universiti Sains Islam Malaysia, Bandar Baru Nilai,

71800 Nilai, Negeri Sembilan, Malaysia

²School of Bioscience and Biotechnology, Faculty of Science and Technology

Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor Darul Ehsan, Malaysia

³Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia

43600 Bangi, Selangor Darul Ehsan, Malaysia

Diserahkan: 12 Disember 2013/Diterima: 10 Jun 2014

ABSTRACT

The aim of this study was to establish a simple, accurate and reproducible method for the identification and quantification of surfactin using high-performance liquid chromatography (HPLC). Previously reported method of identification and quantification of surfactin were time consuming and requires a large quantity of mobile phase. The new method was achieved by application of Chromolith® high performance RP-18 (100 × 4.6 mm, 5 μm) as the stationary phase and optimization of mobile phase ratio and flow rate. Mobile phase consisted of acetonitrile (ACN) and 3.8 mM trifluroacetic acid (TFA) solution of 80:20 ratio at flow rate of 2.2 mL/min was obtained as the optimal conditions. Total elution time of the obtained surfactin peaks was four times quicker than various methods previously reported in the literature. The method described here allowed for fine separation of surfactin in standard sample (98% purity) and surfactin in fermentation broth.

Keywords: High-performance liquid chromatography (HPLC); isocratic; mobile phase; stationary phase; surfactin

ABSTRAK

Tujuan kajian ini adalah untuk menyediakan satu kaedah yang mudah, tepat dan boleh diulang untuk mengenal pasti dan mengkuantifikasi surfaktin menggunakan kromatografi cecair berprestasi tinggi (HPLC). Sebelum ini dilaporkan kaedah kenal pasti dan kuantifikasi surfaktin memerlukan masa yang panjang dan amaun fasa bergerak yang banyak. Kaedah baru telah diperoleh dengan menggunakan kolum Chromolith® high performance RP-18 (100 × 4.6 mm, 5 μm) sebagai fasa pegun dan pengoptimuman nisbah dan kadar aliran fasa bergerak. Fasa bergerak terdiri daripada cecair asetonitril (ACN) dan 3.8 mM asid trifluroacetic (TFA) dengan nisbah 80:20 pada kadar aliran 2.2 mL/min sebagai kadar optimum. Masa analisis yang diperoleh adalah empat kali lebih pantas daripada kaedah yang dilaporkan sebelum ini. Kaedah yang diguna pakai di sini dapat memisahkan piawai surfaktin (98% ketulenan) dan surfaktin dalam sampel fermentasi dengan baik.

Kata kunci: Fasa bergerak; fasa pegun; isokratik; kromatografi cecair prestasi tinggi (HPLC); surfaktin

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*Pengarang untuk surat-menyurat; email: m.hafez@usim.edu.my