Sensitive bioassay of bupivacaine in human plasma by liquid-chromatography-ion trap mass spectrometry.
A sensitive high-performance liquid chromatography-tandem mass spectrometric (HPLC-MS-MS) method, using an ion trap spectrometer, was developed for quantitation of bupivacaine in human plasma. Bupivacaine and an internal standard (ropivacaine) were extracted in a single step from 100 microL of alkalinized plasma with diethyl-ether. The mobile phase consisted of acetonitrile with 0.1% formic acid (50:50, v/v), and was delivered at a flow rate of 0.3 mL/min. The effluent was detected by MS-MS in positive ion mode. Ionisation was performed, using an electrospray ion source, operating at 200 degrees C. The selected reaction monitoring transitions m/z 289-->m/z 140 and m/z 275-->m/z 126 were chosen for bupivacaine and ropivacaine, respectively. Calibration curves were linear over the concentration range of 3.90-500 microg/L with determination coefficients >0.996. The method is accurate (bias <10%) and reproducible (intra-assay and inter-assay precision <15%), with a quantitation limit of 3.90 microg/L, using only 100 microL of plasma. The high specificity and sensitivity, achieved by this fast method (total run-time <3 min), allowed the determination of bupivacaine plasma levels in pediatric patients, following epidural administration of bupivacaine.
Hoizey
Guillaume
G
Laboratoire de Pharmacologie et Toxicologie, Hôpital Maison Blanche, CHU de Reims, 45, rue Cognacq-Jay, 51092 Reims Cedex, France. ghoizey@chu-reims.fr
Lamiable
Denis
D
Robinet
Arnaud
A
Ludot
Hugues
H
Malinovsky
Jean-Marc
JM
Kaltenbach
Matthieu L
ML
Binet
Laurent
L
Boulanger
Christian
C
Millart
Hervé
H
D000097:Acetonitriles / Q000737:chemistry; D000577:Amides / Q000032:analysis / Q000493:pharmacokinetics; D000779:Anesthetics, Local / Q000032:analysis / Q000737:chemistry; D001681:Biological Assay / Q000379:methods*; D002045:Bupivacaine / Q000032:analysis* / Q000737:chemistry* / Q000493:pharmacokinetics*; D002138:Calibration; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D004345:Drug Industry / Q000379:methods; D005561:Formates / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D000077212:Ropivacaine; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D013696:Temperature; D013997:Time Factors
D000097:Acetonitriles; D000577:Amides; D000779:Anesthetics, Local; D005561:Formates; C030544:formic acid; D000077212:Ropivacaine; D002045:Bupivacaine; C032159:acetonitrile