Method validation study of hypoglycin A determination in ackee fruit.

A study was conducted to validate the performance characteristics of a published method entitled "Reversed-Phase Liquid Chromatographic Detection of Hypoglycin A in Canned Ackee Fruit Sample." Hypoglycin A (HG-A) was extracted from ackee fruit with 80% ethanol-water, centrifuged, and filtered; the sample extract then was reacted with phenylisothiocyanate. HG-A was separated by reversed-phase chromatography as the phenylthiocarbamyl derivative and detected at the low nanogram level using a UV detector at 254 nm. A study was conducted to determine recovery of HG-A added to a control ackee fruit sample. A control sample containing a low level of HG-A was spiked with 403.2, 201.6, 96.8, and 48.4 microg HG-A/g ackee fruit, respectively. Twelve replicates were analyzed for each spike level. The mean percent recovery +/- standard deviation for spike levels 403.2, 201.6, 96.8, and 48.4 microg HG-A/g were 94.37 +/- 1.27, 99.12 +/- 2.09, 107.95 +/- 5.42, and 129.18 +/- 15.32%, respectively. The percent coefficient of variation (%CV) for spike levels 403.2, 201.6, 96.8, and 48.4 microg HG-A/g were 1.35, 2.11, 5.02, and 11.86%, respectively. The recovery data indicate that HG-A can be recovered from ackee fruit with excellent accuracy and precision. Precision data obtained from replicate assays of ackee fruit naturally contaminated with low, medium, and high HG-A levels is presented.