1 0 6 n hcl up to 24 hours at 4 u00b0 c step 2 2 0 m cacl 2 up to 24 hours at 4 u00b0 c step 3 0 5 m edta up to 24 hours at 4 u00b0 c step 4 8 0 m licl for 4 hours at 4 u00b0 c and step 5 sterilized h 2 o for 4 hours at 55 u00b0 c bone powders prepared by the method described below were subjected to the following procedure for isbg extraction step 1 0 6 n hcl up to 24 hours at 4 u00b0 c step 2 2 0 m cacl 2 up to 24 hours at 4 u00b0 c step 3 0 5 m edta for 4 hours at 4 u00b0 c and step 4 sterilized h 2 o for 4 hours at 55 u00b0 c instead of using chloroform and methanol solution as described by urist a washing procedure was developed to remove lipids and bone marrow cells in the tissue for protocols 2 and 3 according to the present invention in the step prior to treatment with hcl the bone powder was washed with normal saline at 40 45 u00b0 c for 5 minutes using this procedure 80 of lipids and bone marrow cells were removed at the end of washing the bone powder rinsed with saline appeared to be clear moist and not overly dry as is bone powder treated with a solution of chloroform and methanol to eliminate non crucial chemicals a series of experiments was conducted to.