Development of a novel HPLC‐MS/MS method for the determination of aconitine and its application to in vitro and rat microdialysis samples

Abstract

A sensitive and selective LC-MS/MS method was developed and validated for the determination of aconitine in microdialysate and rat plasma. Extraction of plasma sample was conducted by use of 1% trichloracetic acid and acetonitrile solution with 10 ng/mL internal standard (propafenone) spiked. Microdialysates were analyzed without sample purification. After sample preparation, 2 microL were injected and separated with an isocratic mobile phase consisting of acetonitrile:0.1% formic acid (60:40, v/v) at a flow rate of 0.3 mL/min. The Agilent G6410A triple quadrupole LC/MS system was operated under the multiple-reaction monitoring mode (MRM) using the electrospray ionization technique in positive mode. Overall, the assay exhibited good precision and accuracy. The diffusion properties of aconitine investigated in in vitro microdialysis experiments revealed unfavourable concentration dependence avertable by keeping a constant pH 5.77 using isotonic phosphate buffer solution as perfusate. The mean relative recoveries were 48.23% [coefficient of variation (CV 4.47%)] and 55.38% (CV 2.89%) for retrodialysis and recovery experiments, respectively. The in vivo recovery of aconitine was 34.48% (CV 3.05%) and was stable over the 6 h study period. Following characterization of aconitine both in vitro and in vivo microdialysis, the developed setting is suitable for application in pharmacokinetics and pharmacodynamics studies.

Pharmacokinetics of aconitine in rat skin after oral and transdermal gel administrations

“…The plasma was separated, frozen, and stored at −25°C until assayed. A validated HPLC/MS/MS method was used to determine the aconitine concentration in plasma and dialysis samples (Zhang, et al , ).…”

“…Microdialysis probe recovery was calibrated by triplicate measurements of in vitro recovery, delivery and in vivo delivery (Zhang et al , ) Microdialysis probes were placed in a 200 mL beaker containing well‐stirred isotonic PBS (containing Na 2 HPO 4 2.39 g/L, NaH 2 PO 4 9.34 g/L, NaCl 4.79 g/L, pH 5.77) at three different concentrations of aconitine (50, 200 and 500 ng/mL) at 37°C. The probes were perfused with blank isotonic phosphate buffer (pH 5.77) at 2 μL/min and samples of the dialysate were collected at 20 min intervals; aconitine concentrations were analyzed by LC‐MS/MS to give the relative in vitro recovery.…”

Intestinal Transport of Pure Diester-type Alkaloids from an Aconite Extract across the Caco-2 Cell Monolayer Model

“…Various alkaloids with properties such as analgesic, anti-inflammatory, antiepileptic, local anesthetic, hypotensive, and spasmolytic activities have been made responsible for their pharmacological effects [3,4]. However, these alkaloids are compounds with cardiotoxic and neurotoxic activities and show a narrow therapeutic index [5,6]. Therefore, the improper use of Aconitum will result in a risk of severe intoxication [7,8].…”

Crosstalk the Microdialysis in Scientific Research: from Principle to its Applications

“…Zhang et al [101] used the microdialysis coupled with the LC-MS/MS technology to determine the aconitine drug concentrations in the rats plasma and in vitro transdermal skin dialysate. In this study and found that the investigation aconitine inter-day and intra-day stability were good, the in vivo recovery of the probe was (34.48 ± 3.05)%.…”