SURFACE RESPONSE METHOD FOR SEPARATION OPTIMIZATION OF ERITROMYCIN AND KANAMYCIN USING HPLC WITH FMOC-CL DERIVATIZATION
Background and aims: Erythromycin is a macrolide antibiotic and kanamycin is aminoglycoside antibiotic. Due to the lack of chromophoric groups, analysis of erythromycin and kanamycin preceded by derivatization using FMOC-CI. FMOCCl has reactivity to hydroxyl group on erythromycin and reactivity to a...
Saved in:
| Main Author: | |
|---|---|
| Format: | Theses |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/79020 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Background and aims: Erythromycin is a macrolide antibiotic and kanamycin is aminoglycoside antibiotic. Due to the lack of chromophoric groups, analysis of erythromycin and kanamycin preceded by derivatization using FMOC-CI. FMOCCl has reactivity to hydroxyl group on erythromycin and reactivity to amine on kanamycin. Because of the differences in reactivity, it is possible getting two different optimum conditions for kanamycin and erythromycin. So that the optimization is aimed for the separation process. Analysis of erythromycin and kanamycin carried out simultaneously by HPLC so that the optimum conditions required to obtain a good separation value, which is devoted by resolution value R 1,5. Method: Derivatization was performed by the addition of 50gL boric buffer and 150gL FMOC-CI into 50gL of each standard. Analysis using Inertsil@ C8 column, fluorescence detector , composition of the mobile phase water: acetonitrile 36: 64, flow rate of 1.2 ml /min. Surface Response Method used to find the optimum condition, using Minitab software version 16.Factorial Design with 3 factors and 2 level is used as experimental design. Level based on the optimum value in experiments one factor at the time. Factor or independent variable and the value level is A pH of boric buffer (7.3 to 7.5), B concentration of FMOC-CI (4.0-6.0 mg / mL), and C time reaction (40-50 minutes). Results: Empirical equation = -17,742 + 3,050A + O,012B O,OIOC stating the relationship between the independent variables and the response as the value of R resolution. Application of response surface method showed that the optimum resolution value (4.845) was obtained when pH of boric buffer 7.5, FMOC-CI concentration 5gg/mL and time reaction 40 minutes.
|
|---|