High pressure liquid chromatographic determination of strychnine, using a reverse phase solvent system

A simple, rapid extraction and subsequent determination for strychnine, using high pressure liquid chromatography (HPLC) with a reverse phase solvent system, is described. Stomach contents or grain bait containing strychnine were made alkaline with sodium hydroxide and extracted with chloroform. Extract filtrates were injected directly into a liquid chromatograph without further preparation except for dilution, if necessary. Peaks were resolved within 3.5 min and peak heights were used for quantitation. HPLC of strychnine was carried out on a 30 cm X 4 mm id stainless steel column packed with micronBondapak C18. The solvent program was 0.005M phosphate buffer-methanol (60+40) at a flow of 1.5 ml/min. Recovery from spiked stomach contents was 93.9 +/- 3.5%. The detection capability for strynchnine, using the 254 nm ultraviolet detector, was 5 ng. The strychnine peak was collected and subjected to thin layer chromatography with strychnine standards for confirmation.