Analysis of Impurities of Bacterium Source of Standard Endotoxin by Gas Chromatography/Mass Spectrometry

Abstract

To analyse the impurity of bacterium source of standard endotoxin, 3-hydroxy fatty acid species in different endotoxin standards was determined by gas chromatography/mass spectrometry (GC/MS) using N, O-bis (trimethylsilyl) trifluoroacetamide as the silanizing reagent. GC/MS analysis was performed using a gas chromatograph equipped with a 60 m x 0.25 mm i. d. DB-5 fused silica capillary column and an injector at 250 degrees C. Helium was used as the carrier gas under a constant pressure of 206 kPa. The oven was programmed at a rate of 5 degrees C /min from 90 degrees C (held for 5 min) to 280 degrees C (held for 5 min). The sample size was 1 microL. The transfer line was kept at 280 degrees C. The quadrupole mass spectrometer was operated in electron impact (EI) ionization mode, and the temperature of the source was kept at 250 degrees C. The kind of 3-hydroxy fatty acids in 9 000 EU/tube national standard endotoxin, 20 EU/tube working standard endotoxin, Escherichia coli, Pseudomonas aeruginosa and deionized water were determined to study the purity of bacterium source of the standard endotoxin. It was shown that 9 000 EU/tube endotoxin standard and Escherichia coli only contained 3-hydroxytetradecanoic acid. There was 3-hydroxydodecanoic acid in 20 EU/tube working standard endotoxin, which indicated the presence of impurity of bacterium source.