title: Development and validation of a high-performance liquid chromatographic assay using solid-phase extraction for the novel antitumor agent pancratistatin in human plasma.
abstract: 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.
authors.affiliation: Paterson Institute for Cancer Research, Section of Drug Development and Imaging, Withington, Manchester, UK. pkhan@picr.man.ac.uk
authors.forename: P
authors.initials: P
authors.lastname: Khan
authors.forename: S
authors.initials: S
authors.lastname: Abbas
authors.forename: B
authors.initials: B
authors.lastname: Petit
authors.forename: R
authors.initials: R
authors.lastname: Caffrey
authors.forename: V
authors.initials: V
authors.lastname: Megram
authors.forename: A
authors.initials: A
authors.lastname: McGown
mesh_terms: D047151:Amaryllidaceae Alkaloids; D000972:Antineoplastic Agents, Phytogenic / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007546:Isoquinolines / Q000097:blood*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
chemical_list: D047151:Amaryllidaceae Alkaloids; D000972:Antineoplastic Agents, Phytogenic; D007546:Isoquinolines; C044997:pancratistatin