High-performance liquid chromatographic method for the determination of nefazodone and its metabolites in human plasma using laboratory robotics

doi:10.1016/0378-4347(91)80207-S

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 570, Issue 1, 18 September 1991, Pages 129-138

https://doi.org/10.1016/0378-4347(91)80207-S Get rights and content

Abstract

A quantitative analytical method, using high-performance liquid chromatography and ultraviolet detection, has been established for the determination of nefazodone (NEF) and its metabolites, m-chlorophenylpiperazine (mCPP), p-hydroxynefazodone (PHN), and hydroxynefazodone (HO-NEF), in human plasma. The fully automated, robotic procedure consisted of addition of internal standard (aprindine), extraction with butyl chloride, followed by phase separation, organic phase evaporation, reconstitution of the residue, and injection onto the chromatographic system. The limits of detection for NEF, mCPP, PHN, and HO-NEF were 5, 1, 10, and 5 ng/ml, respectively, at a signal-to-noise ratio of 4. The method had a linear range of 10–1000 ng/ml for NEF and HO-NEF, 20–2000 ng/ml for PHN, and 2.5–250 ng/ml for mCPP. Correlation coefficients of 0.996 or greater were obtained during validation and study sample analysis.

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Cited by (49)

Separation and determination of chlorophenylpiperazine isomers in confiscated pills by capillary electrophoresis
2013, Journal of Pharmaceutical and Biomedical Analysis
Citation Excerpt :
For the same biological samples, liquid chromatography using electrochemical (HPLC–ECD) [11], and ultraviolet (HPLC–UV) [12] detection has also been applied. Human plasma has been inspected for mCPP by HPLC–UV [13–17] and LC–MS [18]. The isomer has also been determined by HPLC–UV in human plasma, red blood cells [19] and plasma and breast milk [20].
A simple capillary electrophoretic method with spectrophotometric UV detection at 236 nm has been developed for the selective separation and determination of 1-(2-chlorophenyl)piperazine (oCPP), 1-(3-chlorophenyl)piperazine (mCPP) and 1-(4-chlorophenyl)piperazine (pCPP) in confiscated pills. Several cyclodextrin derivatives were tested to compose the background electrolyte (BGE). The optimized BGE contained 20 mmol/L phosphoric acid adjusted to pH 2.5 with triethylamine and 10 mmol/L α-cyclodextrin, which provided acceptable resolution of analytes and candidate interferents in less than 15 min. The analyses were performed at constant voltage of 25 kV in 60 cm (effective length 50 cm; 50 μm i.d.) uncoated fused-silica capillary maintained at 25 °C with sample injection at 4826 Pa for 8 s. Procaine at a concentration of 0.1 mg/mL was used as internal standard (IS). Possible interference from other drugs such as amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, 3,4-methylenedioxy-N-ethylamphetamine, 1-(3-trifluoromethylphenyl)piperazine and cocaine was also examined. The analytical curves were linear (R² = 0.9994–0.9995) in the range of 10–200 μg/mL (for oCPP and mCPP) and 20–200 μg/mL for pCPP. Limits of detection (LODs) were 2.0 μg/mL (oCPP), 2.5 μg/mL (mCPP) and 3.5 μg/mL (pCPP). Intraday precision at three concentration levels and six replicates of each level (10, 100, 200 μg/mL of each analyte; n = 18) was evaluated for the corrected peak area ratio of analyte to IS and the migration times giving RSDs ≤ 4.9%. The accuracy was estimated for mCPP by a recovery test at the same three concentration levels and recoveries varied from 101.0 to 101.6%. The method has been successively applied to the analysis of 17 confiscated pills based mostly on mCPP.
High-throughput quantitation of nefazodone and its metabolites in human plasma by high flow direct-injection LC-MS/MS
2007, Journal of Pharmaceutical and Biomedical Analysis
A rapid, selective and sensitive high-performance liquid chromatography tandem mass spectrometry (LC–MS/MS) method coupled with high flow direct-injection on-line extraction has been developed and validated for the simultaneous quantitation of nefazodone and its three active metabolites, hydroxynefazodone, triazole-dione (BMS-180492) and m-chlorophyenylpiperazine (mCPP) in human plasma. The method utilized d₇-nefazodone, d₇-hydroxynefazodone, d₄-BMS-180492 and d₄-mCPP as internal standards (IS). The plasma samples were injected into the LC–MS/MS system after simply adding the internal standard solution and centrifuging. The required extraction and chromatographic separation of the analytes were achieved on an Oasis^® HLB column (on-line extraction column, 1 mm × 50 mm, 30 μm) and a conventional Luna C8 column (analytical column, 4.6 mm × 50 mm, 5 μm). Detection was by positive ion electrospray tandem mass spectrometry. The total analysis run time for each sample was 2 min, which included the time needed for on-line extraction, chromatographic separation and LC–MS/MS analysis. The assay was validated for each analyte and the concentrations ranged from 2.0 to 500 ng/ml for nefazodone, hydroxynefazodone and mCPP and from 4.0 to 1000 ng/ml for BMS-180492, respectively. The assay was used for the high-throughput sample analysis of thousands of pharmacokinetic study samples and was proven to be rapid, accurate, precise, sensitive, specific and rugged.
Rapid and simple methods for quantitative analysis of some antidepressant in pharmaceutical formulations by using first derivative spectrophotometry and HPLC
2003, Farmaco
Two rapid, simple and accurate first derivative spectrophotometry and HPLC method for the determination of nefazodone hydrochloride and sertraline hydrochloride in pharmaceutical formulations are discussed. The first one is a derivative spectrophotometric procedure and the second one is based on a HPLC method with a UV detector. In the first method, first derivative spectrophotometry, nefazodone hydrochloride or sertraline hydrochloride by measurement of their first derivative signals at 241.8–256.7 nm (peak-to-peak amplitude), or 271.6–275.5 nm (peak-to-peak amplitude), respectively. Calibration graphs were established for 10.0–42.0 μg ml⁻¹ nefazodone hydrochloride, or 8.0–46.0 μg ml⁻¹ sertraline hydrochloride. In the other method, HPLC, the UV detection was carried out at 265.0 nm (nefazodone hydrochloride) and 270.0 nm (sertraline hydrochloride). The samples were chromatographed on a Supercosil RP-18 column. The mobile phases were methanol:acetonitrile:phosphate buffer at pH 5.5 (10:50:40 v/v/v) (nefazodone hydrochloride) and methanol:phosphate buffer at pH 4.5 (20:80 v/v) (sertraline hydrochloride). The results obtained from first derivative spectrophotometric method were comparable with those obtained by using HPLC. It was concluded that both the developed methods are equally accurate, sensitive, and precision could be applied directly and easily to the pharmaceutical formulations of nefazodone hydrochloride and sertraline hydrochloride, respectively.
Electrochemical characterisation of nefazodone hydrochloride and voltammetric determination of the drug in pharmaceuticals and human serum
2002, Analytica Chimica Acta
Citation Excerpt :
The widespread use of this compound and the need for clinical and pharmacological study require fast and sensitive analytical techniques to determine the presence of the drug in pharmaceutical formulations and serum samples. Up to now the most common techniques for the determination of the drug in commercial dosage form and biological fluids have been based on HPLC with UV detection [4–6] or with mass spectrometric detection [7,8] or coulometric detection [9] methods, but potentiometric method with ion-selective membrane electrode [10] has also been used. Electrochemical methods have proved to be very sensitive for the determination of organic molecules, including drugs and related molecules in pharmaceutical dosage forms and biological fluids [11–20].
Nefazodone, an antidepressant was electrochemically studied in various buffer systems and at different pH using glassy carbon electrode. Nefazodone was electrochemically oxidized at all pH values. According to the linear relation between the peak current and the nefazodone concentration differential pulse (DPV) and square wave (SWV) voltammetric methods for its quantitative determination in pharmaceuticals and human serum were developed. For analytical purposes, a very well resolved diffusion controlled voltammetric peak was obtained in 0.1 M H₂SO₄ at 0.99 and 1.03 V for DPV and SWV techniques, respectively. The linear response was obtained in the ranges of 8×10⁻⁷ to 6×10⁻⁴ M with a detection limit of 2.1×10⁻⁷ M for DPV and 1.17×10⁻⁷ M for SWV techniques. The repeatability and reproducibility of the methods were within 1.03, 0.81% relative standard deviations (R.S.D.) for peak currents and 0.40, 0.20% R.S.D. for peak potentials, for DPV and SWV, respectively. Precision and accuracy of the developed method was checked by recovery studies. The proposed methods were successfully applied to the individual tablet dosage form and human serum.
LC determination and purity evaluation of nefazodone HCl in bulk drug and pharmaceutical formulations
2001, Journal of Pharmaceutical and Biomedical Analysis
A simple, selective and reproducible reversed-phase liquid chromatography (LC) method has been developed for the quantitative determination of nefazodone hydrochloride (I) in the presence of its related impurities, namely 5-ethyl-4-(2-phenoxyethyl)-2H-1,2,4-triazol-3-(4H)one (II), 1-(3-chlorophenyl)-4-(3-chloropropyl) piperazine hydrochloride (III) and 1,1¹-trimethylene-bis[4-(3-chlorophenyl) piperazine] hydrochloride (IV). The separation was achieved using an Inertsil ODS-3V (250×4.6 mm²) column and a mobile phase comprising 0.05 M KH₂PO₄ (pH 3.0), acetonitrile and methanol in the ratio 50:40:10 (v/v/v). The method has been completely validated and proven to be rugged. The limit of detection and limit of quantification for impurities II, III and IV were found to be 50, 79 and 91 ng/ml, and 152, 240 and 280 ng/ml, respectively. The intra- and inter-day assay precision of the method was within 1.2% relative standard deviations. The developed method was applied to the pharmaceutical dosage form (Tablet, Serzone-R) and the percentage recoveries ranged from 99.1 to 100.7. The percentage recovery of impurities ranged from 96.2 to 108.9. The stability studies were performed for nefazodone solution placed on laboratory bench and in the refrigerator for 60 days. The method was proved to be stability indicating in solution.
Nefazodone pharmacokinetics depressed children and adolescents
2000, Journal of the American Academy of Child and Adolescent Psychiatry
To describe the pharmacokinetics and safety of nefazodone (NFZ) in depressed children and adolescents.
Depressed youths aged 7 to 17 years were eligible to participate. Intensive sampling for pharmacokinetic analyses of NFZ and 3 of its active metabolites was performed after single and multiple dose administration. Treatment was continued for 6 more weeks and titrated to maximize clinical response.
Twenty-eight patients were enrolled. Systemic exposure to NFZ and 3 metabolites was generally higher in children than adolescents. NFZ and metabolite disposition profiles showed high intra- and interpatient variability. Compared to published data in adults, the half-life of NFZ and 2 of its metabolites appears shorter in children and adolescents. Meta-chlorphenylpiperazine pharmacokinetic parameters were different in 5 patients determined to be poor metabolizers of cytochrome P450 2D6 (CYP2D6). NFZ was well tolerated, and administration was associated with significant reductions (p < .001) in depressive symptoms.
The pharmacokinetics of NFZ in pediatric patients is highly variable. NFZ appears to be safe in this small, short-term study. Pediatric patients who are poor metabolizers of CYP2D6 do not appear to be at increased risk for NFZ-associated adverse events. Open-label treatment of NFZ is associated with reductions in depressive symptoms.

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High-performance liquid chromatographic method for the determination of nefazodone and its metabolites in human plasma using laboratory robotics

Abstract

Soc. Neurosci. Abstr.

Soc. Neurosci. Abstr.