According to the formula of loading capacity,
loading capacity= [(Total amount of drug-Free amount of drug)/nanoparticles weight] x 100.
1. how do you determine the total amount of drug, free amount of drug and nanoparticles weight?
2. I have obtained a standard curve from the ELISA test. The dilution factor is 5 and the dilution of standards is in pg/ml. Based on the y=mx+c equation, I am able to find the concentration of the supernatant. However, I am not sure the unit of it? is it pg/ml as well?
1. how do you determine the total amount of drug, free amount of drug and nanoparticles weight?
From the uv calibration curve of your drug you will get amount of drug loaded by subtracting total amount of drug by Free drug. This total amount is the initial concentration of drug you have added in your chitosan nanoparticles and the free drug can be find out by (supernatant absorbance + intercept/ slope).
2. I have obtained a standard curve from the ELISA test. The dilution factor is 5 and the dilution of standards is in pg/ml. Based on the y=mx+c equation, I am able to find the concentration of the supernatant. However, I am not sure the unit of it? is it pg/ml as well?
The unit of concentration you have found from the calibration curve is same the concentration of drug solution you prepared for calibration curve study i.e. pg/ml.
After finding free drug from above formula you have to multiply that free drug concentration by dillution factor to get your actual free drug in supernatant.
Now you need amount of you nanoparticles in the formulation you can get it by freez dryng nps and and weigh the pellet.
DRUG LOADING CAPACITY = (total drug weight -free drug weight) /weight of nps(g)
for % * by 100
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