Hi all, i prepared a solution of CIP 100mg/100ml (as stock solution) which concentration is 1g/L, which i dissolve in 0.01N HCl solution. now i prepared with different concentrations like 10mg/L 1ml from stock solution and 99 ml distilled water, similarly 20mg/L,30mg/L..50mg/L 5ml from stock solution and 95ml distilled water, but when i check it for absorbance at 278 or 277nm wavelength it goes above 4.00 on UV spectrometer. i doubled checked all my solutions, methods and dilutions, UV instrument all is working well. kindly i need some guidance.
Thanks in advance
Hello,
an OD greater than 1 when reading on a spectrophotometer means that the solution is saturated and therefore this result should not be taken.
Obviously, your solution would have to be less concentrated so that the spectro values are less than 1, however I have a question for you, your 0 (the control or the blank), how did you prepare it? What does it contain?
in the blank I have 0.01N HCl solution, I added 0.86ml of 36% HCl to 100 ml distilled water
your blank is not exactly correct because in your solutions you add distilled water and therefore you dilute the 0.01 N of HCl, which you do not do for the blank, that is to say you leave the HCl at 0.01N. For each concentration, try to prepare its blank and look at the results, at the same time further dilute the solutions, try for example the concentrations 1mg/L and 0.1mg/L and see if the OD values are less than 1.
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