- High Performance Liquid Chromatography Hplc Experiment Determination Of Ambroksol Hci Methyl Paraben And Propyl Para 1 (88.83 KiB) Viewed 59 times
what is the pulse-free output? How did we achieve it?
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) Experiment: Determination of Ambroksol HCI, Methyl Paraben and Propyl Paraben in a commercial syrup qualitatively by using High Performance Liquid Chromatography (HPLC). Solutions (Already Prepared): Separate Standards: 10 mg/l. Ambroksolici, 10 mg/l Methyl Paraben, and 10 mg/l, Propyl Paraben standards (already prepared). Mobile Phase: Methanol/0.5% ammonium acetate buffer (60/40), pH adjusted to 5.5. Column: Thermo Scientific Acclaim, C18 Procedure: 1. Tum on the pump and UV absorbance detector. 2. Adjust the flow rate to 1.0 mL/min. 3. Wait until the detector reads a constant absorbance value, if it is different than 0.000 press the autozero button. 4. Take a few milliliters of 10 mg/L Ambroksol HCL solution with syringe. Be careful that the solution inside the syringe must not contain any air bubbles; if there is any it should be removed. 5. Place the syringe into the injection port when it was at the load position and inject the solution from syringe until you see few droplets of the solution coming out of the stainless steel pipe. 6. Then, turn the knob to the inject position and at the same time initiate data acquisition. First the system peak will appear on the chromatogram. Then, the analyte peak will be observed. From the chromatogram find the maximum point of the peak and read the retention time of the analyte. 7. Repeat the same procedure for the Methyl Paraben and Propyl Paraben solutions to find their retention times. 8. Inject an unknown mixture and identify the peaks. 9. Calculate number of plate and theoretical plate height for each ion injected alone.