How can I correctly obtain derivative absorbance values at zero-crossing wavelengths using UVWin software for simultaneous estimation of two drugs?

Rajshekher Upadhyay @Rajshekher-Upadhyay

22 April 2025 0 9K Report

I am working on the simultaneous estimation of two drugs using zero-crossing UV-derivative spectroscopy. I have recorded the zero-order spectra and generated 1st to 4th order derivative spectra using UVWin software. After identifying suitable zero-crossing wavelengths for each drug, I prepared a series of standard dilutions for calibration curve construction.

Now, I have a specific doubt:

To measure the response (absorbance or amplitude) at the zero-crossing wavelength for each standard concentration, should I use the Simple Read or Advanced Read function? Do I need to convert each spectrum into the required derivative order before recording the amplitude, or is reading from the zero-order spectrum sufficient?

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