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Green HPLC-DAD and HPTLC Methods for Quantitative Determination of Binary Mixture of Pregabalin and Amitriptyline Used for Neuropathic Pain Management.
- Source :
-
Journal of chromatographic science [J Chromatogr Sci] 2021 May 20; Vol. 59 (6), pp. 536-547. - Publication Year :
- 2021
-
Abstract
- First analytical methods were herein developed for determination of pregabalin (PGB) and amitriptyline (AMT) as an active binary mixture used for management of neuropathic pain whether in pure forms or in human biological fluids (plasma/urine). First method is green high-performance liquid chromatography-diode array detector (HPLC-DAD) after derivatization of PGB with ninhydrin (NIN) on a reversed-phase C18 column using a mobile phase consisting of ethanol:water (97:3%, v/v) pumped isocratically at 0.8 mL/min; AMT were scanned at 215 nm, whereas PGB-NIN was scanned at 580 nm. Second method is High-performance thin-layer chromatography (HPTLC), where PGB and AMT were separated on silica gel HPTLC F254 plates, using ethanol:ethyl acetate:acetone:ammonia solution (8:2:1:0.05, by volume) as a developing system. AMT peaks were scanned at 220 nm, whereas PGB peaks were visualized by spraying 3% (w/v) ethanolic NIN solution and scanning at 550 nm. Linear calibration curves were obtained for human plasma and urine spiked with PGB and AMT over the ranges of 5-100 μg/mL and 0.2-2.5 μg/band for PGB, and 1-100 μg/mL and 0.1-2.0 μg/band for AMT for HPLC-DAD and HPTLC methods, respectively. The suggested methods were validated according to Food and Drug Administration guidelines for bioanalytical methods validation and they can be applied for routine therapeutic drug monitoring for the concerned drugs.<br /> (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Subjects :
- Amitriptyline urine
Analgesics, Non-Narcotic urine
Anti-Anxiety Agents urine
Drug Monitoring methods
Humans
Limit of Detection
Neuralgia drug therapy
Pregabalin urine
Amitriptyline blood
Analgesics, Non-Narcotic blood
Anti-Anxiety Agents blood
Chromatography, High Pressure Liquid methods
Chromatography, Thin Layer methods
Pregabalin blood
Subjects
Details
- Language :
- English
- ISSN :
- 1945-239X
- Volume :
- 59
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Journal of chromatographic science
- Publication Type :
- Academic Journal
- Accession number :
- 33778855
- Full Text :
- https://doi.org/10.1093/chromsci/bmab031