高效毛细管区带电泳测定内毒素

Determination of lipopolysaccharide by capillary zone electrophoresis]

OBJECTIVE: To detect lipopolysaccharides (LPS) in frangibility bacilli, control the procedure of LPS preparation, examine the ally of the product by high performance capillary zone electrophoresis (CZE), and obtain rapidly high alloy of LPS for scientific research. METHODS: Theoptyline was added to the sample as an internal standard. The LPS was separated and detected by CZE in sodium tetraborate-borate buffer. The concentration of the buffer was 30 mmol.L-1 (pH 8.0); the diameter of the capillary column was 75 microns, and the length was 57 cm; the CZE operative voltage was 25 kV, the electric current was 112 microA, and the detecting wavelength was 230 nm. RESULTS: CZE was first internally used to separate and detecte LPS in bacillus. The LPS had a good linearity in the range of 2-30 mg.L-1 (r = 0.996). The average recovery rate was 99%-101%, the in-day and day-to-day coefficients of variation (CV) were less than 5%, and the minimum detectable quantity was 1.0 mg.L-1. CONCLUSION: Compared with the limulus test (LT) and gel electrophoresis, this method can resist heat source interference and is more simple, rapid and accurate. Moreover, it can control the preparative process of LPS immediately, and may be an advanced method to determine LPS.