Malaysian Journal of Analytical Sciences Vol 21 No 6 (2017): 1352 - 1365

EFFECT OF BUFFER COMPOSITION ON THE ANALYSIS OF LIPOPOLYSACCHARIDES FROM Escherichia coli 055:B5 AND UT189 BY CAPILLARY ELECTROPHORESIS WITH DIRECT UV DETECTION

(Kesan Komposisi Penimbal ke atas Analisis Lipopolisakarida dari Escherichia coli 055:B5 dan UT189 oleh Elektroforesis Kapilari dengan Pengesan UV)

Fun Man Fung¹, Min Su¹, Sam Fong Yau Li^1,2*

¹Department of Chemistry,

National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore

²Singapore-Peking-Oxford Research Enterprise (SPORE) Programme,

NUS Environmental Research Institute (NERI) #02-01, T-Lab Building (TL), 5a Engineering Drive1 1, Singapore 117411, Singapore

*Corresponding author: chmlifys@nus.edu.sg

Received: 7 November 2016; Accepted: 18 September 2017

Abstract

Endotoxins are known to many as bacterial toxins. Conventional analysis of endotoxins by capillary electrophoresis-mass spectrometry is laborious and time consuming. A faster capillary electrophoresis method using direct ultraviolet (UV) detection was performed to analyze endotoxins isolated from Escherichia coli (E. coli) 055:B5 and UT189. In this study, we compared different running buffers and buffer additives to examine the effects of each on the electropherograms obtained. The optimum buffer system was then used to analyze of lipopolysaccharides (LPS) signals from the bacteria E. coli 055:B5 (source strain CDC 1644-70), E. coli K-235 (source strain ATCC 13027) and Klebsiella pneumoniae (source strain ATCC 15380). The data generated for each variant peak showed very good precision and high reproducibility for migration time with percentage RSD values of ≤1.2% (n = 3). Similarly, the percentage RSD for peak areas were ≤5.3% (n = 3) across the three LPS.

Keywords: capillary electrophoresis, direct UV, endotoxin, lipopolysaccharides, buffer

Abstrak

Endotoksin diketahui sebagai bakteria yang berbahaya. Kaedah konvensional bagi endotoksin oleh elektroforesis kapilari-jisim spektrometri adalah bersifat rumit dan memerlukan tempoh masa yang panjang. Kaedah elektroforesis kapilari yang pantas menggunakan pengesan ultralembayung telah dijalankan untuk menganalisa endotoksin yang dipisahkan dari Escherichia coli (E. coli) 055:B5 and UT189. Dalam kajian ini, larutan penimbal yang berbeza telah diuji dan penambahan penimbal dilakukan untuk mengkaji kesan bagi setiap elektropherogram yang terhasil. Sistem penimbal yang optimum kemudian digunakan untuk menganalisa respon lipopolisakarida (LPS) dari bakteria E. coli 055:B5 (sumber strain CDC 1644-70), E. coli K-235 (sumber strain ATCC 13027) and Klebsiella pneumoniae (sumber strain ATCC 15380). Data yang terhasil bagi puncak setiap varian menunjukkan ia adalah jitu dan kebolehulangan yang tinggi bagi masa pemindahan dengan nilai peratus RSD ialah ≤1.2% (n = 3). Begitu juga, peratus RSD bagi luas – luas puncak ketiga – tiga LPS ialah ≤5.3% (n = 3).

Kata kunci: elektroforesis kapilari, UV langsung, endotoksin, lipopolisakarida, penimbal

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