Method Development for the Analysis of Pharmaceuticals with Acethylcholinesterase Activity in Water Using HPLC-DAD and Solid Phase Extraction

https://doi.org/10.22146/ijc.21263

Samuel Budi Wardhana(1*), Marit Kolb(2), Müfit Bahadir(3), Bambang Purwono(4), Tutik Dwi Wahyuningsih(5)

(1) Department of Chemistry, Semarang State University, Kampus Sekaran, Gunungpati, Semarang 50229, Central Java
(2) Institute of Environmental and Sustainable Chemistry, Technische Universität Braunschweig, Hagenring 30, D-38106 Braunschweig
(3) Institute of Environmental and Sustainable Chemistry, Technische Universität Braunschweig, Hagenring 30, D-38106 Braunschweig
(4) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Jl. Sekip Utara, Yogyakarta 55281
(5) Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Gadjah Mada, Jl. Sekip Utara, Yogyakarta 55281
(*) Corresponding Author

Abstract


An SPE followed by HPLC-DAD method with ion pair chromatography technique to analyze pharmaceuticals with acethylcholinesterase activity including pyridostigmine (PYR), galathamine (GAL), neostigmine (NEO), eserine (ESE), and donepezil (DON) in water samples was developed. Acetylcholinesterase (AChE) inhibitors have been used to treat less severe dementias such as Alzheimer’s disease. Chromatographic separation was achieved using reversed-phase SymmetryShield column using gradient system with mobile phase consisting of H2O/ACN (99:1, v/v) as mobile phase A with 10 mM sodium 1-hexanesulfonate and 0.1% acetic acid (HAc). The HPLC/DAD method was linear between concentrations of 5 to 100 ng/μL. The IDL and IQL ranged from 0.50 to 1.25 ng/μL and 1.5 to 3.0 ng/μL, respectively. SPE was used to extract and clean up the target substances in spiked pure water, tap water, and wastewater samples. The application of extraction method of 5 target substances in wastewater sample was divided into 2 parts: Oasis WCX (6 mL, 500 mg) for PYR and Oasis HLB (6 mL, 200 mg) for GAL, NEO, ESE and DON. The developed SPE and HPLC/DAD method is applicable for quantification of the 5 target substances in water samples in a concentration range > 50 µg/L and assumable lower for DON (> 25 µg/L).

Keywords


acetylcholinesterase; solid phase extraction; ion pair chromatography

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DOI: https://doi.org/10.22146/ijc.21263

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Indonesian Journal of Chemisty (ISSN 1411-9420 / 2460-1578) - Chemistry Department, Universitas Gadjah Mada, Indonesia.

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