A simple way for extracting singular porphyrin peaks from total fluorescence spectrum of plasma
,1,* Hamid keshvari
,2 Zahra hassannejad
,3 Mohammad hassan emami
1. Gastrointestinal and Hepatobiliary Diseases Research Center
2. Faculty of Biomedical Engineering
3. Pediatric Urology and Regeneration Medicine Research Center
4. Gastrointestinal and Hepatobiliary Diseases Research Center, MUI
Porphyrins are organic compounds formed from the breakdown of hemoglobin. any disturbance in porphyrins’ level can be a sign of an illness, like “porphyria” which is a famous metabolic disorder. therefore there’re many reports around finding an easier method to determine porphyrin level in blood. current research wants to introduce a simple way for extracting the discriminant porphyrin peaks from total plasma fluorescence spectrum.
Plasma samples were collected, prepared and fluorescence spectroscopies were done for any of the following sample/set-up variants: extractor solvent, storing temperature, excitation wavelength, excitation slit, fluorometric cuvette volume, pmt voltage, scan speed. then the optimum preliminary conditions were chose.
finally the fluorescence spectroscopy of the samples were done at the optimum procedure. moreover the samples were excited at the flavin adenine dinucleotide (fad) excitation maxima.
For any of the supposed variant, the fluorescence spectrum was illustrated and based on the “porphyrin peak discrimination criterion” the optimum spectroscopy conditions were chosen. the 450nm is fad excitation maxima and has no overlap with any other plasma’s fluorophors excitation spectrum. so we can reach the singular porphyrin peaks by means of easily subtracting 450nm excited spectrum from main spectrum.
In conclusion, we propose to use the following variants to gain a better porphyrin diagnosis: analytical grade ethanol as an extraction solvent, storing samples at 4°c before fluorometry, excitation wavelength of around 400nm, 10nm slit width, 400µl cuvette, middle pmt voltage and normal scan speed.
Fluorescence spectroscopy, porphyrin detection, plasma porphyrin level, singular porphyrin peak