Sensitive Analysis of Epilepsy Drug, Phenobarbital, Based on Column Type Solid Phase Extraction and HPLC-DAD System
Abstract
An analytical approach has been developed for the sensitive determination of Phenobarbital, which is used as an antiepileptic drug molecule. The analysis of this active molecule, which has a very limited study even for its direct determination in the literature review, is mostly done with highly complex device systems such as LC-MS. In analysis with conventional HPLC systems, the limit of detection cannot be in most cases lower than 1 µg mL-1 With this study, a separation and preconcentration method based on solid phase extraction (SPE) was developed for trace phenobarbital molecules, so that even very low concentrations could be monitored. In the proposed method, the target molecules were enriched with column type SPE method, and then their analysis were carried out by with the HPLC-DAD system. As an SPE sorbent, a polymeric material, poly(ethylene glycol dimethacrylate-N-methacryloyl-L-tryptophan methyl ester) [poly(EGDMA-MATrp)], was used in extraction experiments. Experimental variables such as pH of medium, type and amount of desorption solvent, electrolyte effect have been studied and optimized step by step. The linear working range under the optimized conditions were determined in the range of 10.00-400.00 ng mL-1 with the limit of detection as 3.57 ng mL-1. Quantitative results were obtained in recovery experiments with the help of model solutions including phenobarbital molecule
Keywords
Phenobarbital, Solid phase extraction, HPLC, Analytical method development
References
- [1] Vermeij T. A. C., Edelbroek P. M., Robust isocratic high performance liquid chromatographic method for simultaneous determination of seven antiepileptic drugs including lamotrigine, oxcarbazepine and zonisamide in serum after solid-phase extraction, Journal of Chromatography B., 857(1) (2007) 40-46.
- [2] Velayati S., Saadati F., Shayani-Jam H., Shekari A., Valipour R., Yaftian M. R., Fabrication and evaluation of a molecularly imprinted polymer electrochemical nanosensor for the sensitive monitoring of phenobarbital in biological samples, Microchemical Journal, 174 (2022) 107063.
- [3] Mobed A., Shirafkan M., Charsouei S., Sadeghzadeh J., Ahmadalipour A., Biosensors technology for anti-epileptic drugs, Clinica Chimica Acta, 533 (2022) 175-182.
- [4] Lyseng-Williamson K. A., Levetiracetam: a review of its use in epilepsy, Drugs, 71 (2011) 489-514.
- [5] A Spectrophotometric Method for the Determination of Nitrite and Nitrate, Eurasian J. Anal. Chem., (2017).
- [6] Jacob S., Nair A. B., An updated overview on therapeutic drug monitoring of recent antiepileptic drugs, Drugs in R&D, 16 (2016) 303-316.
- [7] Ventura S., Rodrigues M., Pousinho S., Falcão A., Alves G., Determination of lamotrigine in human plasma and saliva using microextraction by packed sorbent and high performance liquid chromatography–diode array detection: An innovative bioanalytical tool for therapeutic drug monitoring, Microchemical Journal, 130 (2017) 221-228.
- [8] D’Urso A., Locatelli M., Tartaglia A., Molteni L., D’Ovidio C., Savini F., de Grazia U., Therapeutic drug monitoring of antiseizure medications using volumetric absorptive microsampling: where are we?, Pharmaceuticals, 14(7) (2021) 627.
- [9] Sultan M., Simultaneous HPLC determination and validation of amphetamine, methamphetamine, caffeine, paracetamol and theophylline in illicit seized tablets, Int. J. Pharm. Pharm. Sci., 6(4) (2014) 294-298.
- [10] Pena A., Pelantova N., Lino C. M., Silveira, M. I. N., & Solich, P., Validation of an analytical methodology for determination of oxytetracycline and tetracycline residues in honey by HPLC with fluorescence detection, Journal of Agricultural and Food Chemistry, 53(10) (2005) 3784-3788.