Case: Homicide or suicide?

Before analysis, we need several reagents.

For the Mobile phase,

Mix formic acid of 0.1% (v/v) with acetonitrile (solution 1). Mix formic acid of 0.1% (v/v) with aqueous solution (solution 2). Mix solution 1 with solution 2 to a ratio of (24:76, v/v).

For Urine sample,

We chose to take urine sample instead of serum sample as according to research, aconitines will not retain in serum after 24 hours of ingestion. However, it is still detectable in the urine after 23-62hours.

To prepare the sample, mix one milliliter of urine sample with 2.5ng of Methyllycaconitine (I.S) and 2 mL of 0.0025% (v/v) ammonia solution.

To prepare blank urine sample : Purify from deceased's urine sample using urine purification kits. Removal of nucleic acids and proteins.

For preparatation of standards: mix one mililiter of respective aconitine with 2.5ng of Methyllycaconitine (I.S) and 2 mL of 0.0025% (v/v) ammonia solution.

*Prepare 4 sets of urine sample as test will be run thrice and average result will be taken for a fair test. ( 3 urine samples from deceased, and a blank urine sample as control)

*Prepare 3 standards for each aconitines (aconitine, hypaconitine, jesconitine, mesaconitine) according to the table below: