High-performance liquid chromatographic analysis of the 2-chloroprocaine metabolite, diethylaminoethanol, in blood and serum.
OBJECTIVE
2-Chloroprocaine is rapidly metabolized in the blood to yield 2-chloro-para-aminobenzoic acid (an inactive metabolite) and diethylaminoethanol (DEAE). DEAE possesses local anesthetic activity. The only reported assay for DEAE is a colorimetric method.


METHODS
Clinical samples of whole blood and serum were obtained from patients receiving stepped intravenous infusions of 3% 2-chloroprocaine. A high pH-dependent liquid-liquid extraction step with diethyl ether was used to eliminate interfering peaks in high-pressure liquid chromatography (HPLC) analysis. Separation and quantitation were performed using HPLC on a polymeric-reversed phase column with a mobile phase consisting of 10% or 20% acetonitrile (for whole blood or serum analysis, respectively) in 50 mm aqueous sodium phosphate buffer, pH = 11.50. The elution order of DEAE and its analogues was tested to interpret the HPLC separation mechanism.


RESULTS
Extraction recovery of DEAE from whole blood was 67 +/- 13.5%, from serum, 71 +/- 12.2%, and from water, 75 +/- 2.9%. The high pH value of the mobile phase resulted in sharp, well-resolved peaks with run times of approximately 8 minutes using 20% acetonitrile. The lower limit of detection was 5 ng/mL of DEAE from a 1-mL sample. The elution order of DEAE and its analogues indicated that separation was based on the hydrophobicity of the analytes rather than polar group interactions occurring with silica-based stationary phase.


CONCLUSIONS
A new, simple and rapid HPLC method for extraction and measurement of DEAE in whole blood or serum samples is reported here.
Meng
Q C
QC
Department of Anesthesia, The Center for Anesthesia Research, Hospital of the University of Pennsylvania, Philadelphia, USA.
Kramer
T H
TH
Arthur
G R
GR
Kim
P S
PS
Ferrante
F M
FM
D000779:Anesthetics, Local / Q000008:administration & dosage / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004983:Ethanolamines / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007262:Infusions, Intravenous; D011343:Procaine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood; D015203:Reproducibility of Results
D000779:Anesthetics, Local; D004983:Ethanolamines; D011343:Procaine; C004616:chloroprocaine; C007369:2-diethylaminoethanol