Validation of HPLC-UV Methods for Quantitatively Determining Landiolol and Its Major Metabolite in Human Blood

Abstract

Landiolol hydrochloride is a newly developed cardioselective, ultra short-acting b 1 -adrenergic receptor blocking agent. Landiolol hydrochloride has much higher cardioselectivity (b 1 /b 2 ϭ255) than currently available b 1 -blockers, such as esmolol hydrochloride (b 1 /b 2 ϭ33).1) This drug is rapidly hydrolyzed by both carboxylesterase in the liver and pseudocholinesterase in the plasma 2) to an inactive metabolite (M-1) with a rapid elimination half-life of about 4 min, which is significantly shorter than the elimination half-lives of conventional b 1 -blockers.3) Based on these features, landiolol hydrochloride has been used in the emergency management of atrial fibrillation, atrial flutter and tachycardia as well as in perioperative arrhythmia control.The introduction of a target controlled infusion (TCI) system into clinical practice ensures that a target plasma drug concentration is obtained to induce and maintain anesthesia during perioperative anesthetic management. Based on a pharmacokinetic model, the TCI system determines the initial loading dose needed to achieve a target plasma drug concentration and the infusion rate needed to maintain a constant concentration, and controls the intravenous infusion completely automatically.4) Therefore, application of the TCI system to landiolol hydrochloride administration is expected to allow more effective and safe drug administration.After the quantitation of landiolol in blood, establishing the pharmacokinetic model is necessary for the application of the TCI system. About the quantitation method of landiolol and its major metabolite, although the summarized method was already reported, 5,6) the method details and validation data had not been reported.In this report, validated HPLC-UV methods that quantitatively determine landiolol and its major metabolite in human blood were reported in order to contribute to progress of clinical research of landiolol hydrochloride such as the application of the TCI system. MATERIALS AND METHODS Chemical and ReagentsLandiolol, M-1, an internal standard for landiolol (IS-1), and an internal standard for M-1 (IS-2) were synthesized at Ono Pharmaceutical Co., Ltd. Methanol, ethanol, acetonitrile, ethyl acetate were HPLC January 2009 121 Notes Validation of HPLC-UV Methods for Quantitatively Determining Landiolol and Its Major Metabolite in Human BloodYoshitaka HASHIMOTO,* Sachiko NAGO, Masako TSUNEKAWA, Kimio SHIBAKAWA, and Yasuyuki MIYATA Pharmacokinetic Research Laboratories, Research Headquarters, Ono Pharmaceutical Co., Ltd.; 17-2 Wadai, Tsukubashi, Ibaraki 300-4247, Japan. Received June 24, 2008; accepted October 15, 2008; published online October 16, 2008 Landiolol is an ultra-short-acting b b 1 -adrenergic receptor blocking agent that is used for both perioperative and postoperative patients with tachycardia during general anesthesia. Validated HPLC-UV methods that quantitatively determine landiolol and its major metabolite (M-1) in human blood were reported for clinical research of landiolol. These analytes were...

Detection of landiolol using high-performance liquid chromatography/fluorescence: A blood esterase-sensitive ultra-short-acting β1-receptor antagonist

“…The sample preparation method developed in this study was very simple compared with the previously reported approach [13]. The fluorescence intensity of landiolol was weak and was not much different from that of UV absorption when the authentic compound was analyzed using the previously reported method.…”

“…However, the employment of fluorescence detection in our present method dramatically reduced the chromatographic background signals when the blood sample was analyzed, resulting in the achievement of a sample preparation with 5 times higher sensitivity using a simple purification procedure. For UV detection in the previous method, further complicated purification steps for sample preparation were required [13]; a process which most likely would yield quantitative errata because of the rapid continuous degradation of landiolol after blood sampling (even in cases where blood samples were immediately mixed with ice-cold ethanol after sampling, 10-30% of landiolol were degraded). Thus, the use of enzyme inhibitors, such as neostigmine in the present study, is essential to prevent landiolol degradation, especially when blood is collected in the operation theater.…”

“…1), this drug is thought to be rapidly hydrolyzed to an inactive metabolite by the esterase in blood. Although an analytical method for quantitative determination of landiolol in blood has recently been published [12,13], studies probing on enzymatic degradation after blood sampling have yet to be attempted. Furthermore, a previous method that relies on high-performance liquid chromatography (HPLC) separation with UV detector in quantitative determination of blood landiolol concentrations requires extremely complicated sample preparation procedures with poor detection sensitivity limits exceeding 50 ng/ml [12].…”

Determination of landiolol, an ultra-short-acting β1-receptor antagonist, in human plasma by liquid chromatography–tandem mass spectrometry

“…To date, several methods for the determination of landiolol in human blood have been reported [12,13]. However, based on high performance liquid chromatography (HPLC) with UV or fluorescence detection, these methods have inadequate sensitivity and selectivity.…”

“…In previous assays [12,13], blood samples were immediately mixed with an ethanolic solution of neostigmine to stabilize landiolol prior to sample preparation.…”