title: Comparison of high-performance liquid chromatography and polyclonal fluorescence polarization immunoassay for the monitoring of midazolam in the plasma of intensive care unit patients.
abstract: Midazolam (M) is used as an induction agent for anesthesia. The main metabolite is alpha-hydroxymidazolam (OM), which is pharmacologically active. Use of M for sedation is a recent application, rapidly gaining favor. Monitoring of the level of sedation is fundamental in that an excessive and prolonged effect is associated with the risk of complications. Thus, it was felt both necessary and useful to measure circulating M levels. We compared a high-performance liquid chromatography (HPLC) assay with fluorescence polarization immunoassay (FPIA) for the measurement of M in the serum of 138 sedated patients in the intensive care unit (i.e., 179 samples). Response of the OM was also assessed. The degree of crossover of the metabolite was between 76.8 and 32.7%. The equation of the regression line for sigma HPLC (i.e., the sum M + OM) versus FPIA was TDx = 1.1585 sigma HPLC + 143.42 (R = 0.966). The 95% confidence interval for the slope was 1.1551, 1.1619. The regression slope differed significantly from 1 (p < 0.001) and shows that FPIA measurements overestimated concentrations obtained by HPLC on the order of 19%. The discrepancy between the two techniques was all the more notable when concentrations were > 1,000 ng/ml. The relative selectivity of Abbott industrial reagent in terms of benzodiazepines leads to the identification of what might be called a midazolam-like (M-like) activity covering both M and OM. The development of a global FPIA method for measurement of this M-like activity in sedated patients provides a satisfactory solution to the question raised.
authors.affiliation: Service de Pharmacie Clinique, Groupe Hospitalier Necker-Enfants Malades, Paris, France.
authors.forename: P
authors.initials: P
authors.lastname: Bourget
authors.forename: V
authors.initials: V
authors.lastname: Bouton
authors.forename: A
authors.initials: A
authors.lastname: Lesne-Hulin
authors.forename: P
authors.initials: P
authors.lastname: Amstutz
authors.forename: M
authors.initials: M
authors.lastname: Benayed
authors.forename: D
authors.initials: D
authors.lastname: Benhamou
authors.forename: P L
authors.initials: PL
authors.lastname: Dufieux
authors.forename: G
authors.initials: G
authors.lastname: Goursot
authors.forename: S
authors.initials: S
authors.lastname: Grosbuis
authors.forename: J P
authors.initials: JP
authors.lastname: Haberer
authors.forename: F
authors.initials: F
authors.lastname: Jardin
authors.forename: P
authors.initials: P
authors.lastname: Kirstetter
authors.forename: J
authors.initials: J
authors.lastname: Marty
authors.forename: A
authors.initials: A
authors.lastname: Mercatello
authors.forename: B
authors.initials: B
authors.lastname: Page
authors.forename: J L
authors.initials: JL
authors.lastname: Pourriat
authors.forename: T
authors.initials: T
authors.lastname: Vassal
mesh_terms: D000328:Adult; D000368:Aged; D002851:Chromatography, High Pressure Liquid; D003422:Critical Care / Q000379:methods; D003429:Cross Reactions; D005260:Female; D016231:Fluorescence Polarization Immunoassay; D006801:Humans; D006993:Hypnotics and Sedatives / Q000097:blood*; D008297:Male; D008874:Midazolam / Q000097:blood*; D008875:Middle Aged; D011446:Prospective Studies
chemical_list: D006993:Hypnotics and Sedatives; D008874:Midazolam