Development and validation of a high-performance liquid chromatographic assay using solid-phase extraction for the novel antitumor agent pancratistatin in human plasma

The stability of the experimental anti-tumour agent pancratistatin in human plasma has been investigated. A solid-phase extraction technique and an HP LC assay with external standards have been developed and validated. Extract ion was performed using C-18 cartridges and HPLC, analysis was performed on a 15 cm Hypersil BDS column using isocratic elution with 13% acetonitrile and aqueous solution of 1% (w/v) acetic acid. The lower limit of quantifica tion for pancratistatin in 5% DMF-95% water was found to be 0.58 ng/ml (+/- 10.58%) and 2.3 ng/ml (+/-9.2%) following extraction from human plasma. Mea n recovery of 89.4% (+/-4.73%) was obtained over the concentration range 0. 0023-9.45 mu g/ml for a five day validation study. Pancratistatin was stabl e at room temperature in light or dark for at least 15 days, in the refrige rator at 4 degrees C for at least 16 days and in the freezer at -20 degrees C or -80 degrees C for at least 28 days. Under all conditions monitored, % recovery of pancratistatin from human plasma was greater than 95% and no e vidence of degradation had occurred. There also was no loss of pancratistat in after three cycles of freezing and thawing. (C) 1999 Published by Elsevi er Science B.V. All rights reserved.