Toxicological analysis of chlorhexidine in human serum using HPLC on a polymer-coated ODS column.
A simple and reliable high-performance liquid chromatographic (HPLC) method for analyzing chlorhexidine in human serum was developed. After the addition of an internal standard, levomepromazine, 0.2 mL serum was deproteinized with 10% perchloric acid. The acidic supernatant was neutralized with 1M potassium carbonate solution, and the insoluble salt was removed by centrifugation. An aliquot of the supernatant was applied to HPLC with UV detection (260 nm). HPLC separation was achieved on a polymer-coated ODS column equilibrated with acetonitrile/water containing 0.05% trifluoroacetic acid, 0.05% heptafluorobutyric acid, and 0.1% triethylamine (40:60, v/v). The calibration curve was linear in the concentration range from 0.05 to 50.0 microg/mL, and the lower limit of detection was 0.05 microg/mL. The accuracy and precision of the method were evaluated at concentrations of 0.5 microg/mL and 5.0 microg/mL. The coefficients of variation ranged from 4.0 to 4.5%. The concentration of chlorhexidine in the serum of a patient who died after a suspected intravenous injection of chlorhexidine gluconate was determined.
Kudo
Keiko
K
Department of Forensic Pathology andSciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Ikeda
Noriaki
N
Kiyoshima
Akiko
A
Hino
Yukiko
Y
Nishida
Naoki
N
Inoue
Naohide
N
D000891:Anti-Infective Agents, Local / Q000097:blood* / Q000506:poisoning; D002710:Chlorhexidine / Q000031:analogs & derivatives* / Q000097:blood* / Q000506:poisoning; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D017809:Fatal Outcome; D005260:Female; D005554:Forensic Medicine / Q000379:methods; D006801:Humans; D008875:Middle Aged; D015203:Reproducibility of Results
D000891:Anti-Infective Agents, Local; C010882:chlorhexidine gluconate; D002710:Chlorhexidine