Synthesis, Characterization and Bioassay Studies of p-Nitroanilide Derivatives as Potential ChromogenicSubstrate for Endotoxin Screening
Maisara Abdul Kadir, Rumaizah Che Zulkifli, Asnuzilawati Asari, Nur Shuhaila Haryani Abdul Haris and Noraznawati Ismail* Author for corresponding; e-mail address: maisara@umt.edu.my
Volume: Vol.46 No.3 (May 2019)
Research Article
DOI:
Received: 4 September 2018, Revised: -, Accepted: 17 December 2018, Published: -
Citation: Kadir M.A., Zulkifli R.C., Asari A., Haris N.S.H.A. and Ismail N., Synthesis, Characterization and Bioassay Studies of p-Nitroanilide Derivatives as Potential ChromogenicSubstrate for Endotoxin Screening, Chiang Mai Journal of Science, 2019; 46(3): 558-567.
Abstract
Five p-nitroanilide derivatives namely, tert-butyl (2-((4-nitrophenyl)amino)-2-oxoethyl)carbamate (1), tert-butyl (1-((4-nitrophenyl)amino)-1-oxopropan-2-yl)carbamate (2), tert-butyl (4-methyl-1-((4-nitrophenyl)amino)-1-oxopentan-2-yl)carbamate (3), tert-butyl (1-((2-((4-nitrophenyl)amino)-2-oxoethyl)amino)-1-oxopropan-2-yl)carbamate (4) and tert-butyl (1-((4-(methylthio)-1-((4-nitrophenyl)amino)-1-oxobutan-2-yl)amino)-1-oxopropan-2-yl)carbamate (5) have been successfully synthesized from combination of protected amino acid and p-nitroaniline. These compounds were fully characterized by combination of spectroscopic techniques such as Fourier Transform Infrared (FTIR), Ultraviolet-Visible (UV-Vis), 1H and 13C Nuclear Magnetic Resonance (NMR) and elemental analysis. Common bioassay method, Enzyme-linked Immunosorbent Assay (ELISA) was used to analyze the potential of these compounds to act as chromogenic substrate for endotoxin. The results showed that all of the compounds (except compound 5) gave positive response when interacted with endotoxin by converting the clear solution into yellow cloudy solution. This study also shows that compound 3 that associated with leucine moieties resulted rapid response towards endotoxin which by far acts as the most promising potential chromogenic substrate.