{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T17:32:20Z","timestamp":1783099940625,"version":"3.54.6"},"reference-count":43,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2021,3,29]],"date-time":"2021-03-29T00:00:00Z","timestamp":1616976000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Taif University Researchers Supporting Project","award":["TURSP-2020\/56"],"award-info":[{"award-number":["TURSP-2020\/56"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,5,20]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>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\u2013DAD) 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\u00a0mL\/min; AMT were scanned at 215\u00a0nm, whereas PGB\u2013NIN was scanned at 580\u00a0nm. 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\u00a0nm, whereas PGB peaks were visualized by spraying 3% (w\/v) ethanolic NIN solution and scanning at 550\u00a0nm. Linear calibration curves were obtained for human plasma and urine spiked with PGB and AMT over the ranges of 5\u2013100\u00a0\u03bcg\/mL and 0.2\u20132.5\u00a0\u03bcg\/band for PGB, and 1\u2013100\u00a0\u03bcg\/mL and 0.1\u20132.0\u00a0\u03bcg\/band for AMT for HPLC\u2013DAD 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.<\/jats:p>","DOI":"10.1093\/chromsci\/bmab031","type":"journal-article","created":{"date-parts":[[2021,2,25]],"date-time":"2021-02-25T17:45:29Z","timestamp":1614275129000},"page":"536-547","source":"Crossref","is-referenced-by-count":10,"title":["Green HPLC\u2013DAD and HPTLC Methods for Quantitative Determination of Binary Mixture of Pregabalin and Amitriptyline Used for Neuropathic Pain Management"],"prefix":"10.1093","volume":"59","author":[{"given":"Ibrahim A","family":"Naguib","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy, Taif University, PO Box 11099, Taif 21944, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nesma A","family":"Ali","sequence":"additional","affiliation":[{"name":"Analytical Toxicology Laboratory, Forensic Medicine Authority, Ministry of Justice, Cairo 11647, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fadwa A","family":"Elroby","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, Forensic Medicine Department, Beni-Suef University, Beni-Suef 62521, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed R","family":"Elghobashy","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, Analytical Chemistry Department, Cairo University, Cairo 11562, Egypt"},{"name":"Faculty of Pharmacy, October 6 University, October 6 city, Giza 12585, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,3,29]]},"reference":[{"key":"2021052106393258000_ref1","doi-asserted-by":"crossref","first-page":"1630","DOI":"10.1212\/01.wnl.0000282763.29778.59","article-title":"Neuropathic pain: Redefinition and a grading system for clinical use: Consensus statement on clinical and research diagnostic criteria","volume":"70","author":"Treede","year":"2008","journal-title":"Neurology"},{"key":"2021052106393258000_ref2","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1016\/S1474-4422(10)70143-5","article-title":"Neuropathic pain: Diagnosis, pathophysiological mechanisms, and treatment","volume":"9","author":"Baron","year":"2010","journal-title":"The Lancet Neurology"},{"key":"2021052106393258000_ref3","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1503\/cmaj.060146","article-title":"Neuropathic pain: A practical guide for the clinician","volume":"175","author":"Gilron","year":"2006","journal-title":"Canadian Medical Association Journal"},{"key":"2021052106393258000_ref4","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1016\/j.pjnns.2014.11.002","article-title":"Diagnosis and management of neuropathic pain: Review of literature and recommendations of the Polish Association for the Study of Pain and the Polish Neurological Society\u2013part two","volume":"48","author":"Szczudlik","year":"2014","journal-title":"Neurologia i Neurochirurgia Polska"},{"key":"2021052106393258000_ref5","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1111\/j.1468-1331.2006.01511.x","article-title":"EFNS guidelines on pharmacological treatment of neuropathic pain","volume":"13","author":"Attal","year":"2006","journal-title":"European Journal of Neurology"},{"key":"2021052106393258000_ref6","doi-asserted-by":"crossref","first-page":"S3","DOI":"10.4065\/mcp.2009.0649","article-title":"Recommendations for the pharmacological management of neuropathic pain: An overview and literature update","volume":"85","author":"Dworkin","year":"2010","journal-title":"Mayo Clinic Proceedings"},{"key":"2021052106393258000_ref7","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1111\/j.1468-1331.2010.02999.x","article-title":"EFNS guidelines on the pharmacological treatment of neuropathic pain: 2010 revision","volume":"17","author":"Attal","year":"2010","journal-title":"European Journal of Neurology"},{"key":"2021052106393258000_ref8","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1097\/ALN.0b013e3181c43103","article-title":"Practice guidelines for chronic pain management: An updated report by the American Society of Anesthesiologists Task Force on Chronic Pain Management and the American Society of Regional Anesthesia and Pain Medicine","volume":"112","author":"American Society of Anesthesiologists Task Force on Chronic Pain Management","year":"2010","journal-title":"Anesthesiology"},{"key":"2021052106393258000_ref9","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.pain.2007.08.033","article-title":"Pharmacologic management of neuropathic pain: Evidence-based recommendations","volume":"132","author":"Dworkin","year":"2007","journal-title":"Pain"},{"key":"2021052106393258000_ref10","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.amjmed.2009.04.007","article-title":"Treatment of neuropathic pain: An overview of recent guidelines","volume":"122","author":"O\u2019Connor","year":"2009","journal-title":"The American Journal of Medicine"},{"key":"2021052106393258000_ref11","first-page":"89","article-title":"Comparative study of clinical efficacy of amitriptyline and pregabalin in postherpetic neuralgia","volume":"20","author":"Achar","year":"2012","journal-title":"Acta Dermatovenerologica Croatica"},{"key":"2021052106393258000_ref12","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1177\/1049909111412539","article-title":"A comparative efficacy of amitriptyline, gabapentin, and pregabalin in neuropathic cancer pain: A prospective randomized double-blind placebo-controlled study","volume":"29","author":"Mishra","year":"2012","journal-title":"American Journal of Hospice & Palliative Medicine"},{"key":"2021052106393258000_ref13","doi-asserted-by":"crossref","first-page":"63","DOI":"10.4103\/0378-6323.58686","article-title":"Comparative study of clinical efficacy with amitriptyline, pregabalin, and amitriptyline plus pregabalin combination in postherpetic neuralgia","volume":"76","author":"Achar","year":"2010","journal-title":"Indian Journal of Dermatology, Venereology and Leprology"},{"key":"2021052106393258000_ref14","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1111\/j.1464-5491.2009.02806.x","article-title":"Amitriptyline vs. pregabalin in painful diabetic neuropathy: A randomized double blind clinical trial","volume":"26","author":"Bansal","year":"2009","journal-title":"Diabetic Medicine"},{"key":"2021052106393258000_ref15","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.eplepsyres.2006.09.008","article-title":"Pharmacology and mechanism of action of pregabalin: The calcium channel \u03b12\u2013\u03b4 (alpha2\u2013delta) subunit as a target for antiepileptic drug discovery","volume":"73","author":"Taylor","year":"2007","journal-title":"Epilepsy Research"},{"key":"2021052106393258000_ref16","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1053\/eujp.2001.0324","article-title":"Anticonvulsants in neuropathic pain: Rationale and clinical evidence","volume":"6","author":"Jensen","year":"2002","journal-title":"European Journal of Pain"},{"key":"2021052106393258000_ref17","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1080\/10739149.2013.876545","article-title":"Critical review on the analytical methods for the determination of zwitterionic antiepileptic drugs vigabatrin, pregabalin, and gabapentin in bulk and formulations","volume":"42","author":"Kosti\u0107","year":"2014","journal-title":"Instrumentation Science & Technology"},{"key":"2021052106393258000_ref18","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1007\/s10337-011-2004-8","article-title":"Simultaneous determination of pregabalin, sildenafil and its active metabolite in rat plasma utilising SPE followed by LC\u2013MS\u2013MS","volume":"73","author":"D\u017cygiel","year":"2011","journal-title":"Chromatographia"},{"key":"2021052106393258000_ref19","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1365\/s10337-007-0440-2","article-title":"Determination of pregabalin in human plasma using LC-MS-MS","volume":"67","author":"Mandal","year":"2008","journal-title":"Chromatographia"},{"key":"2021052106393258000_ref20","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1097\/FTD.0b013e31827788c0","article-title":"Rapid quantification of gabapentin, pregabalin, and vigabatrin in human serum by ultraperformance liquid chromatography with mass-spectrometric detection","volume":"35","author":"Chahbouni","year":"2013","journal-title":"Therapeutic Drug Monitoring"},{"key":"2021052106393258000_ref21","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.jpba.2010.04.008","article-title":"Liquid chromatographic separation of pregabalin and its possible impurities with fluorescence detection after postcolumn derivatization with o-phtaldialdehyde","volume":"53","author":"Dou\u0161a","year":"2010","journal-title":"Journal of Pharmaceutical and Biomedical Analysis"},{"key":"2021052106393258000_ref22","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/S1570-0232(04)00662-2","article-title":"Simultaneous high-performance liquid chromatographic analysis of pregabalin, gabapentin and vigabatrin in human serum by precolumn derivatization with o-phtaldialdehyde and fluorescence detection","volume":"810","author":"Vermeij","year":"2004","journal-title":"Journal of Chromatography B"},{"key":"2021052106393258000_ref23","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.jchromb.2014.05.030","article-title":"Simultaneous determination of gabapentin, pregabalin, vigabatrin, and topiramate in plasma by HPLC with fluorescence detection","volume":"962","author":"Martinc","year":"2014","journal-title":"Journal of Chromatography B"},{"key":"2021052106393258000_ref24","article-title":"Validated HPLC method for quantification of pregabalin in human plasma using 1-Fluoro-2, 4-dinitrobenzene as derivatization agent","author":"Ahmadkhaniha","year":"2014","journal-title":"Chromatography Research International"},{"key":"2021052106393258000_ref25","first-page":"1","article-title":"Simultaneous estimation of pregabalin and methylcobalamine in pharmaceutical formulation by RP-HPLC method","volume":"4","author":"Bhatt","year":"2013","journal-title":"Journal of Analytical and Bioanalytical Techniques"},{"key":"2021052106393258000_ref26","first-page":"43","article-title":"Quantitative determination of pregabalin by gas chromatography using ethyl chloroformate as a derivatizing reagent in pure and pharmaceutical preparation","volume":"48","author":"Sowjanya","year":"2011","journal-title":"Indian Drugs"},{"key":"2021052106393258000_ref27","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1016\/j.jpba.2012.05.001","article-title":"Development, validation and comparison of two microextraction techniques for the rapid and sensitive determination of pregabalin in urine and pharmaceutical formulations after ethyl chloroformate derivatization followed by gas chromatography\u2013mass spectrometric analysis","volume":"70","author":"Mudiam","year":"2012","journal-title":"Journal of Pharmaceutical and Biomedical Analysis"},{"key":"2021052106393258000_ref28","first-page":"326","article-title":"Simultaneous HPTLC method for estimation of gabapentin and pregabalin","volume":"5","author":"Prasad","year":"2013","journal-title":"International Journal of Pharmacy and Pharmaceutical Sciences"},{"key":"2021052106393258000_ref29","doi-asserted-by":"crossref","first-page":"145","DOI":"10.4172\/2157-7064.1000169","article-title":"Simultaneous estimation of pregabalin and methylcobalamine in pharmaceutical formulation by HPTLC-densitometry method","volume":"4","author":"Shah","year":"2013","journal-title":"Journal of Chromatography and Separation Techniques"},{"key":"2021052106393258000_ref30","doi-asserted-by":"crossref","first-page":"260","DOI":"10.59566\/IJBS.2010.6260","article-title":"Spectrofluorimetric and spectrophotometric determination of pregabalin in capsules and urine samples","volume":"6","author":"Shaalan","year":"2010","journal-title":"International Journal of Biomedical Science: IJBS"},{"key":"2021052106393258000_ref31","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1186\/1752-153X-5-59","article-title":"A novel method for spectrophotometric determination of pregabalin in pure form and in capsules","volume":"5","author":"Bali","year":"2011","journal-title":"Chemistry Central Journal"},{"key":"2021052106393258000_ref32","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.jchromb.2014.09.031","article-title":"Ultrasound-assisted combined with nano-sized molecularly imprinted polymer for selective extraction and pre-concentration of amitriptyline in human plasma with gas chromatography-flame detection","volume":"972","author":"Khanahmadzadeh","year":"2014","journal-title":"Journal of Chromatography B"},{"key":"2021052106393258000_ref33","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1016\/j.jpba.2016.06.027","article-title":"Antidepressants detection and quantification in whole blood samples by GC\u2013MS\/MS, for forensic purposes","volume":"128","author":"Truta","year":"2016","journal-title":"Journal of Pharmaceutical and Biomedical Analysis"},{"key":"2021052106393258000_ref34","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1093\/jat\/21.3.185","article-title":"Selective determination of amitriptyline and nortriptyline in human plasma by HPLC with ultraviolet and particle beam mass spectrometry","volume":"21","author":"Kudo","year":"1997","journal-title":"Journal of Analytical Toxicology"},{"key":"2021052106393258000_ref35","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S0378-4347(96)00193-4","article-title":"Quantitative analysis of amitriptyline and nortriptyline in human plasma and liver microsomal preparations by high-performance liquid chromatography","volume":"685","author":"Ghahramani","year":"1996","journal-title":"Journal of Chromatography B: Biomedical Sciences and Applications"},{"key":"2021052106393258000_ref36","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1016\/j.jpba.2010.04.031","article-title":"Validated LC\u2013MS (ESI) assay for the simultaneous determination of amitriptyline and its metabolite nortriptyline in rat plasma: Application to a pharmacokinetic comparison","volume":"53","author":"Shen","year":"2010","journal-title":"Journal of Pharmaceutical and Biomedical Analysis"},{"key":"2021052106393258000_ref37","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.talanta.2014.10.041","article-title":"A simple dried blood spot method for therapeutic drug monitoring of the tricyclic antidepressants amitriptyline, nortriptyline, imipramine, clomipramine, and their active metabolites using LC-MS\/MS","volume":"134","author":"Berm","year":"2015","journal-title":"Talanta"},{"key":"2021052106393258000_ref38","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1016\/j.saa.2006.01.050","article-title":"Spectrophotometric determination of trazodone, amineptine and amitriptyline hydrochlorides through ion-pair formation with molybdenum and thiocyanate","volume":"65","author":"Mohamed","year":"2006","journal-title":"Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy"},{"key":"2021052106393258000_ref39","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1021\/jf030490p","article-title":"Applications of the ninhydrin reaction for analysis of amino acids, peptides, and proteins to agricultural and biomedical sciences","volume":"52","author":"Friedman","year":"2004","journal-title":"Journal of Agricultural and Food Chemistry"},{"key":"2021052106393258000_ref40","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.chroma.2008.08.021","article-title":"Enantiomeric reversed-phase high-performance liquid chromatography resolution of d-\/l-penicillamine after spirocyclization with ninhydrin and by using copper (II)-l-proline complex as a chiral selector in the mobile phase","volume":"1205","author":"Sotgia","year":"2008","journal-title":"Journal of Chromatography A"},{"key":"2021052106393258000_ref41","author":"US Food and Drug Administration","year":"2001"},{"key":"2021052106393258000_ref42","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jchromb.2013.02.026","article-title":"First HPLC\u2013UV method for rapid and simultaneous quantification of phenobarbital, primidone, phenytoin, carbamazepine, carbamazepine-10, 11-epoxide, 10, 11-trans-dihydroxy-10, 11-dihydrocarbamazepine, lamotrigine, oxcarbazepine and licarbazepine in human plasma","volume":"925","author":"Serralheiro","year":"2013","journal-title":"Journal of Chromatography B"},{"key":"2021052106393258000_ref43","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s40508-016-0051-z","article-title":"Tools and techniques for solvent selection: Green solvent selection guides","volume":"4","author":"Byrne","year":"2016","journal-title":"Sustainable Chemical Processes"}],"container-title":["Journal of Chromatographic Science"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/chromsci\/article-pdf\/59\/6\/536\/37975648\/bmab031.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/chromsci\/article-pdf\/59\/6\/536\/37975648\/bmab031.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,21]],"date-time":"2023-10-21T23:47:21Z","timestamp":1697932041000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/chromsci\/article\/59\/6\/536\/6199107"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,29]]},"references-count":43,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2021,3,29]]},"published-print":{"date-parts":[[2021,5,20]]}},"URL":"https:\/\/doi.org\/10.1093\/chromsci\/bmab031","relation":{},"ISSN":["0021-9665","1945-239X"],"issn-type":[{"value":"0021-9665","type":"print"},{"value":"1945-239X","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2021,7]]},"published":{"date-parts":[[2021,3,29]]}}}