Pengembangan metode analisis obat dengan aktivitas Asetilkolinesterase pada sampel air menggunakan HPLC-DAD dan Solid Phase Extraction; Method Development for the Analysis of Pharmaceuticals with Acethylcholinesterase Activity in Water Using HPLC-DAD and Solid Phase Extraction

Samuel Budi Wardhana Kusuma, Bambang Purwono

2013 | Disertasi | PROGRAM STUDI S2 ILMU KIMIA

An SPE followed by HPLC-DAD method to analyze pharmaceuticals with acethylcholinesterase activity in water samples was developed. Acetylcholinesterase (AChE) inhibitors have been used to treat less severe dementias such as Alzheimer’s disease. In this study, five AChE inhibitors pyridostigmine, galathamine, neostigmine, eserine, and donepezil were selected as target compounds. A HPLC/DAD method with ion pair chromatography technique was used for detection and separation of the target compounds. Effects of the pH of the mobile phase and ion pair reagents in the mobile phase were studied. 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% HAc. The HPLC/DAD method was linear between concentrations of 5 to 100 ng/?L. The IDL and IQL ranged from 0.50 to 1.25 ng/?L and 1.5 to 3.0 ng/?L, respectively. SPE was used to extract and clean up the water samples. An extraction model was conducted with fortification experiments in spiked pure water, tap water, and wastewater samples. The recovery rate of all target substances using WCX cartridge was high for pure and tap water. 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 the extraction and Oasis HLB (6 mL, 200 mg) for GAL, NEO, ESE and DON. The recovery rates of GAL, NEO, ESE and DON using Oasis HLB from wastewater at spiking levels of 50, 125 and 250 ?g/L were in the range of 93.3 to 105.8% with RSD values that ranged between 1.0 and 12.1%. The recovery rates of PYR using Oasis WCX from wastewater samples at the spiking levels of 100 ?g/L were 95.5 (210 nm) and 105.2% (270 nm) and RSDs were 7.2 and 2.6%, respectively. The MQL for GAL, NEO, ESE and PYR (at 210 nm) in wastewater was reached for a concentration of 50 ?g/L and DON at concentration of 25 ?g/L.The developed SPE and HPLC/DAD method is applicable for quantification of the five target compounds in water samples in a concentration range > 50 ?g/L and assumable lower for DON (> 25 ?g/L).

Kata Kunci : asetilkolinesterase; solid phase extraction; kromatografi ion pair