title: Determination of sugammadex in human plasma, urine, and dialysate using a high-performance liquid chromatography/tandem mass spectrometry assay.
abstract: Sugammadex (Bridion®, Merck Sharp & Dohme Corp., Oss, The Netherlands) is a modified γ-cyclodextrin which has the ability to reverse the neuromuscular blockade induced by the steroidal neuromuscular blocking agents rocuronium and vecuronium. The objective of the current study is to describe the bioanalytical methods that have been developed and validated according to US Food and Drug Administration guidelines on bioanalytical method validation, and subsequently applied to determine total sugammadex (i.e., free sugammadex plus sugammadex bound to the neuromuscular blocking agent) in human heparinized plasma, urine and dialysate. Sugammadex was extracted from human plasma and urine using solid phase extraction with Isolute HAX 96-well extraction plates; no extraction was performed on dialysate samples. Samples from plasma, urine, and dialysate were analyzed on a Polaris® C18-A PEEK (polyaryletheretherketone) analytical column (50 mm × 4.6 mm internal diameter, 5 μm) with a linear mobile phase gradient of 0.1% (v/v) formic acid in water:methanol from 70:30 to 20:80. The flow rate was 1 mL/min with a total run time for each injection of 6 min. Tandem mass spectrometric detection was conducted using multiple reaction monitoring under negative ion mode with a turbo ion-spray interface to quantify the concentration of sugammadex. Inter- and intra-assay precision and accuracy were within pre-defined acceptance limits. The presence of rocuronium did not interfere with the assay in plasma, urine or dialysate; similarly, vecuronium did not interfere with the plasma assay (not tested for interference in urine or dialysate). Sugammadex was found to be stable in plasma, urine and dialysate in the short-term at room temperature, in the long-term at -20°C, and after several freeze/thaw cycles. The validated bioanalytical methods developed here have been successfully applied in a series of clinical studies for the determination of total sugammadex in plasma, urine and dialysate.
authors.affiliation: Department of Bioanalytics, Merck Sharp & Dohme Corp., Oss, The Netherlands. marcel.dezwart@merck.com
authors.forename: Marcel A H
authors.initials: MA
authors.lastname: de Zwart
authors.forename: Jolanda
authors.initials: J
authors.lastname: ten Bruggencate-Broeders
authors.forename: Henk J M
authors.initials: HJ
authors.lastname: van Hal
authors.forename: René H J J J
authors.initials: RH
authors.lastname: Megens
authors.forename: Helma W L H
authors.initials: HW
authors.lastname: Frasa
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015314:Dialysis Solutions / Q000032:analysis; D005260:Female; D006801:Humans; D008297:Male; D009466:Neuromuscular Blocking Agents / Q000032:analysis / Q000097:blood / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D000077122:Sugammadex; D053719:Tandem Mass Spectrometry / Q000379:methods*; D047408:gamma-Cyclodextrins / Q000032:analysis* / Q000097:blood / Q000652:urine
chemical_list: D015314:Dialysis Solutions; D009466:Neuromuscular Blocking Agents; D047408:gamma-Cyclodextrins; D000077122:Sugammadex