Detection of Residual Glutaraldehyde in Hemoglobin-Based Oxygen Carrier with High Performance Liquid Chromatography
A method for the determination of glutaraldehyde in hemoglobin-based oxygen carrier(HBOC) by high performance liquid chromatography is described.Free glutaraldehyde was separated from HBOC by 10 kDa ultrafiltration membrane under centrifuged 3000 r/min for 20 min and derivatized under the conditions of pH=1.0,2,4-dinitrophenylhydrazine(DNPH):glutaraldehyde=60:1,reaction time 30 min in 70% acetonitrile medium.Precipitation of derivatives of glutaraldehyde with DNPH was avoided under this condition and it was possible to direct injection of the sample into the LC system.Subsequent reversed phase separation and determination of the derivatives within 20 min with a mobile phase of 70% acetonitile can be achieved.The linear range of this method was from 0.1 to 10.0 mg/L(R2=0.9999) with relative standard deviation of 3.0%(n=6),the standard addition recovery was 95.3%,and the limit of detection was 1.0 ng.