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Simultaneous determination of amiodarone, dronedarone, and their principal metabolites in SD rat plasma by UPLC-MS/MS and its application in pharmacokinetics
⁎Corresponding authors. yexuemei_2009@126.com (Xuemei Ye), xuxuegu1104@126.com (Xuegu Xu)
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Received: ,
Accepted: ,
This article was originally published by Elsevier and was migrated to Scientific Scholar after the change of Publisher.
Abstract
The purpose of the experiment was mainly to establish and verify a precise and straightforward ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) approach for simultaneously analyzing the concentration levels of amiodarone, dronedarone, and their metabolites (desethylamiodarone and desbutyldronedarone) in the plasma of Sprague-Dawley (SD) rats and to investigate the pharmacokinetics of all analytes in SD rats. After rapid protein precipitation by using acetonitrile, we accomplished the chromatographic separation of amiodarone, desethylamiodarone, dronedarone, desbutyldronedarone and ivabradine (internal standard, IS) by using Acquity BEH C18 column and detected through a mass spectrometer with Xevo TQ-S triple quadrupole tandem, choosing the positive ion mode. The approach showed wonderful linearity, and the range of calibration curve for amiodarone was 1–200 ng/mL, desethylamiodarone was 0.1–20 ng/mL, dronedarone was 0.5–100 ng/mL, and desbutyldronedarone was 0.25–50 ng/mL, respectively. For lower limit of quantification (LLOQ), the current method of UPLC-MS/MS can achieve values of 1.0 ng/mL for amiodarone, 0.1 ng/mL for desethylamiodarone, 0.5 ng/mL for dronedarone, and 0.25 ng/mL for desbutyldronedarone, respectively. The accuracy of intra-day and inter-day of all analytes was between −14.8% to 10.9%, while the precision was ≤ 13.3%. For each substance, the recovery rate was > 82.1%, besides, obvious matrix effect was not found. In all conditions, the stability of all analytes was comfirmed to the plasma sample quantification. In addition, the method of UPLC-MS/MS we developed could also be applied to measure the pharmacokinetic characteristics including amiodarone, desethylamiodarone, dronedarone, and desbutyldronedarone in the plasma of SD rats.
Keywords
Amiodarone
Dronedarone
Pharmacokinetics
UPLC-MS/MS
Rat plasma
1 Introduction
Amiodarone (Fig. 1A), an iodinated benzofuran derivative, is a highly lipophilic drug, which has unpredictable pharmacokinetics. Although amiodarone was originally classified as a class III drug which had the ability to prolong refractory in the cardiac field and prevent/terminate re-entry, it showed properties in all four classes of antiarrhythmic drug (AAD), which accounts for nearly 30% of the world market of AAD (Allen LaPointe et al., 2015).
Hepatic cytochrome enzymes CYP3A4 and CYP2C8 can metabolize amiodarone into the active metabolite desethylamiodarone (Fig. 1B). Desethylamiodarone, which can significantly increase the duration of action potentials and reduce the maximum depolarization rate at relevant concentrations clinically (Pallandi & Campbell, 2012; Varro et al., 1996), is the main metabolite of amiodarone and also has antiarrhythmic activity. Amiodarone and desethylamiodarone restrain the activity of some P450 cytochrome enzymes and P-glycoprotein membrane transport systems, which may cause clinically interactions of relevant drugs (Mohamed et al., 2008; Singh, 2006). The use of amiodarone is restricted by the toxic effect and side effects of the parent molecule and desethylamiodarone (Biancatelli et al., 2019). These side effects are manifested in the adverse reactions of the heart, eyes, lungs, liver, dermatology, hematology, psychiatry, thyroid and neuromuscular, and epididymitis and syndrome of inappropriate secretion of antidiuretic hormone can even be caused by chronic amiodarone treatment (Biancatelli et al., 2019). Therefore, as the metabolism is slow and unpredictable, extracardiac toxicity is high and numerous drug interactions, almost one-third of patients cannot maintain long-term treatment for the severe adverse reactions (Allen LaPointe et al., 2015; Dan et al., 2018).
Dronedarone (Fig. 1C), specifically developed for treating atrial fibrillation (AF), is a non-iodinated benzofuran, which was designed to retain the efficacy of amiodarone, with an improved safety profile. The trial conducted in the European Union (EU) in 2009 showed that the group applied dronedarone significantly reduced the composite endpoint of cardiovascular (CV) hospitalization or all-cause death in patients with paroxysmal or persistent AF, compared with Placebo group (Hohnloser, Crijns, Eickels, Gaudin, & Connolly, 2009). Debutyldronedarone (Fig. 1D), formed primarily by CYP3A4, is a major circulating metabolite with similar or even higher plasma exposure than dronedarone. Like amiodarone, dronedarone is not only a substrate of CYP3A4, but also an inhibitor of CYP2D6 and P-glycoprotein, which can affect the pharmacokinetics of many drugs (January et al., 2014; Rosa et al., 2014). Cardiotoxicity and hepatotoxicity of dronedarone are similar to amiodarone. However, due to the absence of iodine in structure and low tissue accumulation, there were no iodine related pulmonary toxicity, ocular toxicity and thyroid toxicity (January et al., 2014; Kozlowski et al., 2012). Therefore, dronedarone is usually used as an alternative to amiodarone. When patients couldn’t tolerate the adverse reactions of amiodarone, dronedarone can be used instead. Given that the long half-life of amiodarone (about 20–100 days for human), when during drug-bridging period, there might be a need to monitor the plasma concentration of amiodarone, dronedarone and their metabolites. And, it is necessary to monitor the plasma concentration of amiodarone and dronedarone clinically, so as to determine which drug caused serious adverse reactions. For these reasons, simultaneous determination of amiodarone and dronedarone in plasma is an ideal strategy for pharmacokinetic and/or toxicokinetic studies and helps in making more accurate decision related to pharmacotherapy.
As far as we know, there have been several reports on the simultaneous determination of biological samples of amiodarone and its main metabolite desethylamiodarone (Kuhn, Gotting, & Kleesiek, 2010; Maes et al., 2006; Shayeganpour, Somayaji, & Brocks, 2007), or dronedarone and its metabolite (Xie, Yang, Zhong, Dai, & Chen, 2011) based on liquid chromatography tandem mass spectrometry (LC-MS/MS) However, only one bioassay simultaneously determined the amiodarone, dronedarone and their main metabolites in plasma, but the pharmacokinetic parameters were not mentioned with long analytical time and complicated extraction procedure (Bolderman, Hermans, & Maessen, 2009).
Thus, there hasn’t been any bioassay which can simultaneously determine the levels of amiodarone, dronedarone and their metabolites (desethylamiodarone and desbutyldronedarone) in plasma and describe their pharmacokinetics based on LC-MS/MS in biological fluids. The aim of the article was to establish an accurate and rapid method for the simultaneous determination of amiodarone, dronedarone and their metabolites in plasma of Sprague-Dawley (SD) rats by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), and to investigate the pharmacokinetics of four analytes from SD rats.
2 Experimental
2.1 Chemicals, materials and reagents
All the analytes of amiodarone, desethylamiodarone, dronedarone, desbutyldronedarone and ivabradine (internal standard, IS, Fig. 1E) were all purity > 98%, and acquired from Beijing Sunflower and Technology Development CO., LTD (Beijing, China), including anlytical reagent (AR) grade formic acid in the study. Acetonitrile and methanol were liquid chromatography (LC) grade and were supplied by Merck Company (Darmstadt, Germany). Water was purified by a Milli-Q Reagent System (Millipore, Bedford, USA).
2.2 Animal experiments
All the six male SD rats (weight 200 ± 20 g), which were fed and drank freely for more than a week in the laboratory with qualified environment, were from the Laboratory Animal Center of Wenzhou Medical University (Zhejiang, China). Rat studies were accomplished on the basis of institutional guidelines abiding by the rules of the Care and Use of Laboratory Animals of Wenzhou Medical University (Zhejiang, China).
After at least of 12 h fasting, 60 mg/kg of amiodarone and 80 mg/kg of dronedarone were orally administered to each rat simultaneously, which were formulated in the solution of 0.5% carboxymethyl cellulose sodium (CMC-Na). Blood was collected from the tail vein (approximately 0.3 mL) at 0, 0.33, 0.67, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 36 and 48 h, then put into a polyethylene tube of 1.5 mL with heparin inside. And then, blood samples were centrifuged (8000g, 4 °C for 10 min). The supernatants were extracted after centrifugation and then put it at −80 °C for the next step of analysis. After monitoring the analyte’s concentration of rat plasma through UPLC-MS/MS, pharmacokinetic parameters were derived using non-compartmental analysis by Drug and Statistics (DAS) 3.0 software (Mathematical Pharmacology Professional Committee of China, Shanghai, China).
2.3 Instrumentations and analytical conditions
The instrumentation applied in the present research was the system of Waters Acquity ultra performance liquid chromatography (UPLC) (Milford, MA, USA), which has an I-CLASS delivery manager of binary solvent, a thermostatted column (set at 40 °C) as well as an autosampler (FTN, set at 10 °C). The Acquity UPLC BEH C18 (2.1 mm × 50 mm, 1.7 μm) column, equipping with a pre-column, was chosen to perform the chromatographic separation. Mobile phase A (acetonitrile) and mobile phase B (0.1% formic acid aqueous solution) were used to separate the analytes completely by an optimized gradient profile. The gradient started from a 0.30 mL/min of the flow rate, 10% acetonitrile for 0.5 min, then increased to 90% acetonitrile in the next 0.5 min; from 1.0 to 2.0 min, gradient was maintained at 90% acetonitrile; from 2.0 to 2.1 min, declined to the starting conditions and kept steady for 1.0 min. The total time was 3.0 min, and the injection volume was a 1.0 µL.
Connected with a source of electro-spray ion (ESI), the MS/MS triple quadrupole system Xevo TQ-S (Milford, MA, USA) was performed in multiple reaction monitoring (MRM) mode using positive ionization. The parameters of the MS system control and collected data were displayed in Table 1 by the Masslynx 4.1 software (Milford, MA, USA).
| Analytes | Precursorion | Production | CV (V) | CE (eV) | RT (min) |
|---|---|---|---|---|---|
| Amiodarone | 646.20 | 100.00 | 10 | 25 | 1.40 |
| Desethylamiodarone | 618.10 | 546.90 | 30 | 18 | 1.37 |
| Dronedarone | 557.50 | 100.10 | 20 | 25 | 1.33 |
| Desbutyldronedarone | 501.00 | 114.00 | 20 | 30 | 1.29 |
| IS | 469.30 | 177.10 | 20 | 25 | 1.18 |
2.4 Preparation of standard and quality control (QC) samples
The corresponding standard substance was dissolved in methanol to obtain the stock solution of the four analytes and IS at the concentration of 1.00 mg/mL, respectively. The stock solutions were diluted with methanol to prepare the mixture of working solution. Similarly, IS working solution was diluted to 50 ng/mL with methanol. Eight non-zero calibration standards were prepared by adding 10 µL of mixed standard working solutions to 90 µL of blank rat plasma. The nominal concentrations of the calibration curve were: amiodarone was 1–200 ng/mL, desethylamiodarone was 0.1–20 ng/mL, dronedarone was 0.5–100 ng/mL, and desbutyldronedarone was 0.25–50 ng/mL, respectively. Prepare quality control (QC) samples in blank plasma samples at four concentration levels: lower limit of quantification (LLOQ), low (LQC), medium (MQC) and high (HQC), respectively, with the levels of amiodarone was 1.0, 2.0, 40, 160 ng/mL, desethylamiodarone was 0.1, 0.2, 4, 16 ng/mL, dronedarone was 0.5, 1.0, 20, 80 ng/mL, and desbutyldronedarone was 0.25, 0.5, 10, 40 ng/mL. All the working and stock fluids were positioned at −80 °C for next step of analysis.
2.5 Extraction procedure
Calibration standards, plasma samples, and quality control samples were prepared as follows: plasma sample of 100 µL, IS working fluid (50 ng/mL) of 20 µL as well as acetonitrile of 300 µL were added into a centrifuge tube of 1.5 mL successively. Then, the mixture was centrifugated for 10 min (13,000g, 4 °C) after being vortexed for 2.0 min. Afterwards, we used a pipette gun to transfer 100 µL supernatant to the autosampler vials, and injected an aliquot of the clear supernatant of 1.0 µL for UPLC-MS/MS analysis.
2.6 Method validation
According to the principles of FDA bioanalysis method validation (Center for Drug Evaluation and Research of the U.S. Department of Health and Human Services Food and Drug Administration, Guidance for industry; Bioanalytical method validation, 2018, http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm064964.htm, Accessed: August 2, 2018.; Tang et al., 2020; Xu et al., 2019), the optimized UPLC-MS/MS method was verified for simultaneously and quantitatively determining the levels of amiodarone, dronedarone, and their metabolites from rat plasma. The related parameters of method validation included selectivity, matrix effect, LLOQ, accuracy and precision, calibration curve, recovery, and stability under all different situations.
The selectivity of the determination was studied by checking whether there was interference between the blank solution (neither IS nor analyte from 6 different rats), real rat plasma and standard solution at the corresponding analytes and IS retention time.
The standard curve was composed of 8 non-zero points of analytes, and the linearity was evaluated. The least-square regression analysis was performed by drawing the ratio of peak area (analyte/IS) against the concentration of labeled plasma. The correlation coefficient (r2) of the corresponding standard curve for each analyte was predicted to be more than 0.99. LLOQ was the lowest concentration point in the calibration curve.
Six duplicate samples were analyzed at LLOQ, LQC, MQC and HQC concentration levels to detect intra-day and inter-day precision and accuracy in three days, consecutively.
The precision was performed as the relative standard deviation (RSD) between repeated measurements, which was less than 15%; the acceptable criteria for accuracy expressed by the relative error (RE) was 85–115%. But the acceptable limits of accuracy and precision at LLOQ should be within 80–120% and RSD should be less than 20%, respectively.
The extraction recovery was defined as: at the same nominal concentration, before and after the extraction, it is the ratio of the mean peak area of analytes in blank matrix. Matrix effect referred to the peak ratio of the spiked analyte to the solvent substituted sample in the blank matrix after extraction at the same nominal concentration. The analytes’ stability at the two QC levels was evaluated by analyzing five duplicate spiked plasma samples in the underlying conditions (Popa et al., 2020): placed under an ambient temperature for 4 h, 4 weeks at −80 °C and 10 °C at the autosampler for 8 h. Freeze-thaw stability was carried out after three entire cycles of freeze and thaw (from −80 °C to room temperature).
3 Results and discussions
3.1 Method development and optimization
By optimizing different separation conditions, high sensitivity, short peak time, high resolution and symmetrical peak pattern were obtained in the experiment. As a result, an Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) column was selected. On the basis of our previous work, satisfying results can be obtained by choosing 0.1% formic acid aqueous and acetonitrile solution to be as mobile phases in LC-MS/MS (Shi, Jiang, Zhang, Tang, & Xu, 2020; Xie, Shi, Chen, Xu, & Ye, 2020). In the developed chromatographic separation, we also found that it was effective and selective to achieve gradient elution with acetonitrile and 0.1% formic acid aqueous solution.
Acetonitrile and methanol are commonly used organic solvents for protein precipitation (PPT). Compared with the traditional sample extraction methods (liquid–liquid extraction and solid phase extraction), PPT is more rapid and convenient, which provides high extraction recovery without obvious endogenous interferences. Compared with methanol (extraction recovery < 80%), acetonitrile was more suitable because of higher extraction recovery (>80%). Therefore, acetonitrile was chosen for PPT.
3.2 Method validation
3.2.1 Selectivity
By comparing the chromatograms of six blank plasma samples from rats in this experiment with plasma samples spiked with standard solution and plasma samples after oral operation, the selectivity of the method was verified. The results were shown in Fig. 2, which revealed that blank plasma did not interfere apparently with the retention time of amiodarone, desethylamiodarone, dronedarone, desbutyldronedarone and IS, at the time of 1.40, 1.37, 1.33, 1.29 and 1.18 min, respectively. As a result, the UPLC-MS/MS method in the experiment was found to be selective and specific.
3.2.2 LLOQ and linearity of calibration curve
In the calibration curve, when the concentration range of amiodarone was 1–200 ng/mL, the concentration range of desethylamiodarone was 0.1–20 ng/mL, the concentration range of dronedarone was 0.5–100 ng/mL, and the concentration range of desbutyldronedarone was 0.25–50 ng/mL, the each curve was highly linear. In all verification runs, the determination coefficient (r2) of linear regression analysis was always greater than 0.99. Table 2 summarized the average equations of the calibration curves of amiodarone, desethylamiodarone, dronedarone and desbutyraldehyde in rat plasma. For LLOQ, the current measured value of amiodarone by using UPLC-MS/MS method was 1.0 ng/mL, desethylamiodarone was 0.1 ng/mL, dronedarone was 0.5 ng/mL, and desbutyldronedarone was 0.25 ng/mL, respectively. The relevant precision and accuracy met requirements of the bioanalytical verification guidelines (within 20%, Table 3).
| Analytes | Regression equation | r2 | Linear range (ng/mL) | LLOQ (ng/mL) |
|---|---|---|---|---|
| Amiodarone | y = 0.0480031x ± 0.0379964 | 0.998 | 1.0–200 | 1.0 |
| Desethylamiodarone | y = 0.078849x ± 0.00692587 | 0.997 | 0.1–20 | 0.1 |
| Dronedarone | y = 0.0609679x ± 0.0390075 | 0.993 | 0.5–100 | 0.5 |
| Desbutyldronedarone | y = 0.0581203x ± 0.00643325 | 0.999 | 0.25–50 | 0.25 |
| Analytes | Concentration (ng/mL) | Intra-day | Inter-day | ||
|---|---|---|---|---|---|
| RSD% | RE% | RSD% | RE% | ||
| Amiodarone | 1.0 | 9.7 | −13.7 | 10.6 | −14.6 |
| 2.0 | 4.5 | −0.4 | 8.5 | −3.2 | |
| 40 | 3.6 | 6.7 | 6.3 | 4.1 | |
| 160 | 2.4 | 4.4 | 3.1 | 2.9 | |
| Desethylamiodarone | 0.1 | 10.1 | −11.3 | 13.3 | −13.8 |
| 0.2 | 5.9 | 1.6 | 6.8 | 1.8 | |
| 4 | 2.6 | 6.8 | 3.6 | 3.7 | |
| 16 | 2.1 | 3.7 | 3.4 | 3.9 | |
| Dronedarone | 0.5 | 5.2 | 9.6 | 6.7 | 3.5 |
| 1.0 | 4.6 | 1.2 | 5.7 | −2.8 | |
| 20 | 4.0 | 2.6 | 2.8 | −1.8 | |
| 80 | 2.5 | 0.0 | 2.6 | −1.8 | |
| Desbutyldronedarone | 0.25 | 9.3 | −14.8 | 11.3 | −14.4 |
| 0.5 | 5.4 | 10.9 | 6.5 | 10.4 | |
| 10 | 4.6 | 2.3 | 5.1 | 3.1 | |
| 40 | 3.0 | −0.2 | 3.1 | 0.4 | |
3.2.3 Accuracy and precision
The accuracy and precision of the intra-day and inter-day of each analyte were presented in Table 3, analyzed at three QC samples (LQC, MQC and HQC). As listed in Table 3, the range of accuracy was within ±15%, and the range of precision was within 15%. UPLC-MS/MS method, newly established for simultaneous quantification of amiodarone, dronedarone and metabolites in rat plasma, was considered to be reliable and highly reproducible.
3.2.4 Recovery and matrix effect
As presented in Table 4, average extraction recoveries in rat plasma of four analytes were 82.1–94.5% at three QC levels, suggesting that the method was highly repeatable. The matrix effect of all analytes in SD rats was 88.0–114.5%, and negligible significant matrix effect was found during data analysis.
| Analytes | Concentration (ng/mL) | Recovery (%) | Matrix effect (%) | ||
|---|---|---|---|---|---|
| Mean ± SD | RSD (%) | Mean ± SD | RSD (%) | ||
| 2.0 | 84.6 ± 11.4 | 13.5 | 102.1 ± 12.6 | 12.3 | |
| Amiodarone | 40 | 91.3 ± 1.7 | 1.9 | 99.6 ± 2.6 | 2.7 |
| 160 | 94.5 ± 8.0 | 8.5 | 88.0 ± 8.9 | 10.1 | |
| 0.2 | 87.4 ± 12.1 | 13.9 | 103.0 ± 8.7 | 8.4 | |
| Desethylamiodarone | 4.0 | 87.9 ± 9.3 | 10.6 | 107.6 ± 11.2 | 10.5 |
| 16 | 92.2 ± 11.1 | 12.0 | 98.7 ± 11.9 | 12.0 | |
| 1.0 | 82.1 ± 9.3 | 11.3 | 109.0 ± 14.2 | 13.0 | |
| Dronedarone | 20 | 87.3 ± 10.3 | 11.8 | 114.5 ± 13.2 | 11.5 |
| 80 | 89.9 ± 8.9 | 9.9 | 108.0 ± 11.4 | 10.5 | |
| 0.5 | 84.7 ± 10.0 | 11.8 | 112.8 ± 11.9 | 10.5 | |
| Desbutyldronedarone | 10 | 88.7 ± 6.3 | 7.1 | 110.6 ± 8.6 | 7.8 |
| 40 | 91.1 ± 9.6 | 10.5 | 111.3 ± 13.5 | 12.1 | |
3.2.5 Stability
Stability of plasma samples was performed under various situations at LQC and HQC concentrations. The results showed that the rat plasma samples remained stable for at least 4 h when they were stored in ambient temperature, at least 28 days when stored at −80 °C, at least 8 h when stored in the autosampler (10 °C) or the three complete circles of freeze and thaw.
3.3 Animal experiments
After an oral dose of 60 mg/kg amiodarone as well as 80 mg/kg dronedarone, the newly established UPLC-MS/MS method was performed to determine the concentrations of plasma of the four analytes in rats. The average concentration of plasma vs time profiles of the four analytes in SD rats were displayed in Fig. 3, and the main pharmacokinetic parameters calculated by DAS 3.0, using non-compartment model analysis, were shown in Table 5.
| Parameters | Amiodarone | Desethylamiodarone | Dronedarone | Desbutyldronedarone |
|---|---|---|---|---|
| AUC0→t (ng/mL•h) | 2638.75 ± 286.93 | 94.80 ± 17.47 | 773.43 ± 316.02 | 415.49 ± 144.16 |
| AUC0→∞ (ng/mL•h) | 2744.86 ± 355.72 | 110.61 ± 15.99 | 782.69 ± 320.23 | 420.03 ± 144.22 |
| MRT0→t (h) | 15.78 ± 2.09 | 17.52 ± 1.48 | 12.70 ± 2.01 | 12.25 ± 1.43 |
| MRT0→∞ (h) | 17.16 ± 3.42 | 22.52 ± 3.66 | 13.22 ± 2.15 | 12.82 ± 1.20 |
| t1/2 (h) | 14.08 ± 5.93 | 17.10 ± 2.99 | 6.93 ± 1.13 | 6.68 ± 1.35 |
| Tmax (h) | 4.67 ± 1.63 | 6.00 ± 0 | 3.17 ± 0.98 | 6.33 ± 3.20 |
| CLz/F (L/h/kg) | 20.80 ± 3.30 | 551.06 ± 71.66 | 118.17 ± 49.63 | 207.87 ± 63.83 |
| Cmax (ng/mL) | 167.28 ± 47.46 | 4.49 ± 1.44 | 55.30 ± 23.74 | 25.88 ± 9.32 |
Two drugs were slowly absorbed after taking a single oral dose of amiodarone and dronedarone, reaching their maximum concentration (Cmax) within 4.67 ± 1.63 h and 3.17 ± 0.98 h post-dose, respectively. Besides, the half-life (t1/2) of amiodarone was 14.08 ± 5.93 h. Although many LC-MS/MS methods for determination of amiodarone and desethylamiodarone in plasma had been found, explicit pharmacokinetic profile and parameters had not been described (Kuhn et al., 2010; Maes et al., 2006; Shayeganpour et al., 2007). As for dronedarone and its metabolite desbutyldronedarone in SD rats, the t1/2 of dronedarone and desbutyldronedarone were 6.93 ± 1.13 h and 6.68 ± 1.35 h, respectively. In addition, Tmax were 3.17 ± 0.98 h and 6.33 ± 3.20 h, respectively. There were few reports on the pharmacokinetics of dronedarone and desbutyldronedarone, and the pharmacokinetic parameters were difficult to be compared in the study. Only one literature determined the pharmacokinetics of amiodarone, dronedarone and their principal metabolites in plasma and myocardium by HPLC, but the sample processing was complex, and no detailed pharmacokinetic parameters were available (Bolderman et al., 2009). However, the half-lives of amiodarone and dronedarone in the drug label are longer than those in the study. These discordances might be due to species differences between humans and rats, as well as individual differences between samples in this study (n = 6).
Therefore, further studies are needed to explore the accurate pharmacokinetic curves and parameters of the four analytes. In short, our study was the first time to establish a UPLC-MS/MS method to simultaneously detect amiodarone, dronedarone and their principal metabolites (desethylamiodarone and desbutyldronedarone) in SD rats, and describe their pharmacokinetic parameters.
4 Conclusions
To summary, it was the first time to establish a UPLC-MS/MS method to simultaneously detect amiodarone, desethylamiodarone, dronedarone and desbutyldronedarone and describe the pharmacokinetics. Moreover, the optimized UPLC-MS/MS method to simultaneously quantify the four analytes in SD rat plasma was more rapid and reliable than the traditional (Bolderman et al., 2009), which could shorten retention time, improve sensitivity and accuracy. This method currently can be used to investigate the pharmacokinetics of the four analytes in SD rats and obtain the pharmacokinetic parameters. When dronedarone is used instead of amiodarone clinically, this method could be performed to detect the concentration levels of four analytes. Besides, the establishment of the UPLC-MS/MS could help identification and screening of drugs when serious adverse reactions occur after taking amiodarone or dronedarone.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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