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Development and validation of a high-performance liquid chromatographic assay using solid-phase extraction for the novel antitumor agent pancratistatin in human plasma.Khan, Parveen; Abbas, Sayara; Petit, B; Caffrey, Robert; Megram, Victoria; McGown, Alan T; Paterson Institute for Cancer Research, Section of Drug Development and Imaging, Withington, Manchester, UK. firstname.lastname@example.org (1999-04-16)The stability of the experimental anti-tumour agent pancratistatin in human plasma has been investigated. A solid-phase extraction technique and an HPLC assay with external standards have been developed and validated. Extraction was performed using C18 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 quantification 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. Mean recovery of 89.4% (+/-4.73%) was obtained over the concentration range 0.0023-9.45 microg/ml for a five day validation study. Pancratistatin was stable at room temperature in light or dark for at least 15 days, in the refrigerator 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 evidence of degradation had occurred. There also was no loss of pancratistatin after three cycles of freezing and thawing.