Calibration curve :

Procedure:
25mg of lopinavir dissolved in 50 ml of acetonitrile . Which gives concentration of 500 g/ml Take 20 ml of above solution and dilute it up to 100ml with acetonitile This will give concentration of 100g/mlwhich will be a stock solution for calibration curve. From that above stock solution withdraw 0.4ml,0 .8ml, 1.2ml,1.6ml,2ml,2.4ml,2.8ml solution and dilute it upto 10 ml of acetonitrile. Above solution will give concentration of 4, 8g 12g, 16g, 20g, 24g, and 28g respectively. Now to perform uv spectroscopy , in spectrometric mode check out the absorption spectra for each and evry solution in range of 200-400 nm. Each spectra are measured atr constant lamda space of 2 and in first derivative method. Following observation are obtained.

Concentration g/ml 4 8 12 16 20 24 28

Abs. -0.111 -0.219 -0.301 -0.426 -0.524 -0.621 -0.724

0 -0.1 -0.2 -0.3 -0.4 -0.5 -0.6 -0.7 -0.8 -0.9 Abs. Linear (Abs.) 0 5 10 15 20 25 30 35

r=0.99944 slope=0.02559 intecept=0.00857

Solubility study : Measurement of extent of solubility of lopinavir in compritol Procedure: 200mg of compritol888(glyceryl behnet) placed in 10ml of beaker. Weigh 10mg of lopinavir Melt this lipid at temp. of above 5c of its melting point M.p.range of compritol is 68-75c .so, preffered temp. is 80c. After complete melting of lipid add slight quantity of lopinavir . Wait until the drug is completely solubilize in melted lipid. Now,add subsequent quantity of lopinavir until the supersaturation
condition .

Now measure the remaining quantity of drug from the total weighed quantity.

Calculation:
weighed quantity of lopinavir- 10mg weighed quantity of compritol -200mg fraction of drug remained at saturation level-5.1mg So, solubilized fraction : total drug-remained drug =10mg-5.1mg =4.9mg So, in 200mg lipid 4.9mg of lopinavir is sobilized. So, in case of compritol lipid obtained ration of lopinavir:compritol was 1:40.

Same procedure was performed for precirol . Obtained ratio of lopinavir:precirol was 1:50