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-mass spectrometric analysis.
The present article reports first time the use of solid-phase microextraction (SPME) and dispersive liquid-liquid microextraction (DLLME) to extract pregabalin (PRG) from urine and pharmaceutical formulations followed by GC-MS analysis after ethyl chloroformate (ECF) derivatization. PRG is an antiepileptic and analgesic drug, which is a structural analogue of γ-amino-butyric acid (GABA). It is approved by Food and Drug Administration (FDA) for the treatment of central nervous system (CNS) disorders and neuropathic pain. Initially PRG was derivatized with ECF in the presence of pyridine at room temperature for 30s. Experimental parameters were investigated for derivatization, SPME and DLLME conditions. The limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.019 μg/ml and 0.063 μg/ml for SPME and 0.022 μg/ml and 0.075 μg/ml for DLLME respectively. The percentage recovery, in case of SPME was in the range of 83-98% while for DLLME it is in the range of 84-98%. The intra and inter-day precisions were found to be less than 6%. The developed methods after ECF derivatization were found to be simple, fast, efficient and inexpensive. DLLME has several advantages like lesser extraction time and cost effectiveness as compared to SPME. The developed methods may find wide application for the routine determination of PRG in biological as well as in quality control samples of pharmaceutical formulations.
Analytical Chemistry Section, CSIR-Indian Institute of Toxicology Research, PO Box 80, M.G. Marg, Lucknow 226001, Uttar Pradesh, India. mohanitrc@gmail.com
Mohana Krishna Reddy
MK
Mudiam
Abhishek
A
Chauhan
Rajeev
R
Jain
Ratnasekhar
R
Ch
Ghizal
G
Fatima
Ekta
E
Malhotra
R C
RC
Murthy
D000700:Analgesics / Q000032:analysis* / Q000652:urine; D000927:Anticonvulsants / Q000032:analysis* / Q000652:urine; D002626:Chemistry, Pharmaceutical; D004304:Dosage Forms; D005563:Formic Acid Esters / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D059627:Liquid Phase Microextraction; D000069583:Pregabalin; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052617:Solid Phase Microextraction; D012997:Solvents / Q000737:chemistry; D013678:Technology, Pharmaceutical / Q000379:methods*; D013997:Time Factors; D016482:Urinalysis; D005680:gamma-Aminobutyric Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000652:urine
D000700:Analgesics; D000927:Anticonvulsants; D004304:Dosage Forms; D005563:Formic Acid Esters; D012997:Solvents; C007658:ethyl chloroformate; D000069583:Pregabalin; D005680:gamma-Aminobutyric Acid