I'm currently working on the degradation of an antibiotic (penicillin). To achieve that, I need a calibration curve. I tried several times but each time I can't get the appropriate absorbance for the prepared dilution (1ppm-10ppm).
I even changed the antibiotic and got the same results.
Any suggestions for this problem?
Should I do a calibration curve using hplc? Up till now I only used a UV-vis spectrophotometer.
#photocatalytic_degradation #antibiotic #calibrarion_curve.
What problem are you exactly having?
Not getting proper correlation coefficient?
Not getting proper repeatability?
Not getting any absorbance?
What wavelength are you using?
Are you getting any peak in the Scan Mode?
What solvent are you using?
Is your UV Vis spectrometer properly calibrated?
I can't obtain logical absorbances, for exemple I get 0.34 for 1ppm and 0.03 or 0 for 10ppm.
The spectrophotometer was calibrated, I used distilled water to dissolve penicillin G.
As the obtained results aren't logical, I can't get a proper correlation coefficient.
I'm using the wavelength 239nm. The wavelength was chosen after the lecture of the absorbance spectre of penicillin g in range 200-800nm
@Emad Hussein that is exactly what I did, unfortunately the obtained absorbances are not good.
I used distilled water to dissolve penicillin and used also distilled water as blank.
The prepared concentrations were 1ppm to 10ppm
Make sure you use QUARTZ glass cuvettes for all standards and samples. Make measurements using the lambda-max wavelength in the UV/VIS range for your sample (There are many variants of penicillin so make sure to choose the correct one for YOUR sample) . Have your teacher/instructor help you set up the instrument before use. Allow plenty of time for the spectrophotometer detector to warm up and equilibrate before use (~ 30 minutes). Balance the detector using a proper blank solution, then measure the sample cuvette. Bracket sample measurements with standard solutions to monitor and detect any changes. Record the data obtained from a complete set of standards, plot the data and compare to your sample. **Your sample will need to be 100% pure for this type of measurement to have any validity (otherwise, HPLC separation and sample prep may be needed).
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