Characterization of Blighia sapida seed extracts.

Blighia sapida, commonly known as the ackee, is a member of the Sapindaceae family. The tree is native to the forests of West Africa and was brought to the Caribbean and later Florida, where it is cultivated as an orchard crop in some areas. Arilli of the fruit are processed to make canned ackee in brine whereas the seeds, raphe and pods are discarded. Physiochemical studies were conducted on extracts of the seed. Qualitative analysis detected the presence of phenolics and reducing sugars. Aqueous extracts of the seeds (ASE) exhibited free radical scavenging activity and had an inhibitory concentration of 2.59 mg/mL. Gas chromatography mass spectrometry led to the identification of several metabolites including amino acids and fatty acids. Hypoglycin B was isolated utilizing ion exchange chromatography. Fourier transform infrared spectroscopy of hypoglycin B detected a band resonating at 3070 cm-1 which may be attributed to the methylenecyclopropane moiety of hypoglycin B. The seeds had a lipid content of 5.72 ± 0.25 % (w/w). The ackee seed oil (ASO) had a saponification value of 152.07 ± 37 and a carotenoid content of 23.7 ± 1.8 mg/kg. The ackee seeds are a source of bioactive components.

Ackee (Blighia sapida K.D. Koenig) Leaves and Arils Methanolic Extracts Ameliorate CdCl2-Induced Oxidative Stress Biomarkers in Drosophila melanogaster.

Different ethnomedical benefits have been documented on different parts of Ackee (Blighia sapida); however, their roles in ameliorating oxidative damages are not well established. CdCl2 inhibitory effects on some oxidative-stress biomarkers and ameliorative potentials of Ackee leaves (AL) and arils (AS) methanolic extracts were studied using Drosophila melanogaster as a model. One to 3-day-old D. melanogaster flies were orally exposed to different concentrations of CdCl2 in their diet for 7 days. The fly's survival profile and negative geotaxis assays were subsequently analysed. Methanolic extracts of AL and AS treatments showed negative geotaxis behaviour, and extracts were able to ameliorate the effect of Cd2+ on catalase and GST activities and increase total thiol and GSH levels, while it reduced the H2O2 generation (p ≤ 0.05) when compared to the control. Furthermore, Cd2+ exhibited noncompetitive and uncompetitive enzyme inhibition on catalase and GST activities, respectively, which may have resulted in the formation of Enzyme-substrate-Cd2+ transition complexes, thus inhibiting the conversion of substrate to product. This study, thus, suggests that the Cd2+ mechanism of toxicity was associated with oxidative damage, as evidenced by the alteration in the oxidative stress-antioxidant imbalance, and that the AL and AS extracts possess essential phytochemicals that could alleviate possibly deleterious oxidative damage effects of environmental pollutants such as CdCl2. Thus, Ackee plant parts possess essential phytonutrients which could serve as valuable resources in heavy metal toxicity management.

Phytochemical screening of aqueous leaf extract of Blighia sapida K.D. Koenig (Sapindaceae) and its analgesic property in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Blighia sapida is traditionally used in treating intercostal pain, psychosis, stomach ache, back pain, and skin diseases. However, there is limited information on the scientific basis for its use traditionally in the treatment of pain. AIM OF STUDY: To identify the major constituents in the aqueous leaf extract of Blighia sapida (AEBS) and to assess its analgesic properties in mice. MATERIALS AND METHODS: Bioactive compounds were identified and quantified in AEBS by High Performance Liquid Chromatography/Photodiode Array Detector (HPLC/DAD). Analgesic activity of AEBS was assessed at doses of 125, 250, and 500 mg/kg p.o., using animal models. RESULTS: Chlorogenic acid, saponins, tannins, caffeic acid, quercetin, gallic acid, pyrogallol, quinine, caffeine, and nicotine were identified. At doses 250 mg/kg (p < 0.05) and 500 mg/kg (p < 0.01), AEBS significantly inhibited acetic acid induced writhing in comparison with the control. It also significantly inhibited pain in the inflammatory phase of the formalin induced paw licking test at 250 mg/kg (p < 0.01) and 500 mg/kg (p < 0.05) doses, in comparison with the control. It did not inhibit pain in the neurogenic phase of the formalin paw licking and in the hot plate tests. CONCLUSION: Blighia sapida leaf extract possesses analgesic activity that is mediated by peripheral mechanisms but not through central mechanisms.

Bioactive micro-constituents of ackee arilli (Blighia sapida K.D. Koenig).

Ackee (Blighia sapida K. D. Koenig) is an exotic fruit widely consumed in the Caribbean countries. While there is extensive research on the presence of hypoglycin A, other bioactive compounds have not been studied. We identified and quantified the changes in bioactive molecules (total phenol, ascorbic acid, hypoglycin A, squalene, D: A-Friedooleanan-7-ol, (7.alpha.), and oleic acid), antioxidant potential, and volatile compounds during two stages of ripe. A clear reduction in hypoglycin A, ascorbic acid, and total polyphenols during the maturation process were observed. On the contrary, oleic acid, squalene, and D: A-Friedooleanan-7-ol, (7.alpha.) contents increased about 12, 12, and 13 times, respectively with advancing maturity. These bioactive molecules were positively correlated with radical scavenging (DDPH and ABTS). Solid phase microextraction (SPME) and gas chromatography coupled mass spectrometry (GC/MS) analysis revealed more than 50 compounds with 3-penten-2-one and hexanal as the major compounds in the fully ripe stage. The results suggested that ripe ackee arilli could serve as an appreciable source of natural bioactive micro-constituents.

Hederagenin glycosides from the fruits of Blighia unijugata.

A phytochemical investigation of Blighia unijugata led to the isolation of eleven hederagenin glycosides. Among these compounds, six are previously undescribed, two are described in their native forms for the first time and three are known whereas firstly isolated from Blighia unijugata. The structure of the undescribed compounds was elucidated on the basis of 2D NMR and mass spectrometry analyses as 3-O-ß-D-xylopyranosyl-(1 → 3)-α-L-arabinopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin, 3-O-ß-D-xylopyranosyl-(1 → 3)-α-L-arabinopyranosyl-(1 → 4)-3-O-acetyl-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin, 3-O-ß-D-glucopyranosyl-(1 → 3)-α-L-arabinopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin, 3-O-ß-D-xylopyranosyl-(1 → 3)-ß-D-xylopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin, 3-O-ß-D-xylopyranosyl-(1 → 3)-ß-D-xylopyranosyl-(1 → 4)-3-O-acetyl-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin, 3-O-α-L-arabinopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin 28-O-ß-D-glucopyranosyl-(1 → 6)-ß-D-glucopyranosyl ester, 3-O-α-L-arabinopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin 28-O-ß-D-glucopyranosyl ester and 3-O-ß-D-xylopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosylhederagenin 28-O-ß-D-glucopyranosyl ester. These results revealed the existence of several conserved structural features that could be used as chemotaxonomic markers for the Blighia genus such as the glycosidic sequence 3-O-α-L-arabinopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranosyl, the occurrence of 3-O-acetylated ß-D-glucopyranosyl units and the systematic presence of hederagenin as aglycone.

Blighia sapida K.D. Koenig: A review on its phytochemistry, pharmacological and nutritional properties.

ETHNOPHARMACOLOGICAL RELEVANCE: Ackee plant (Blighia sapida K. D. Koenig) (Sapindaceae) is used in Sub-Saharan Africa (where it has its origin) and in different parts of the world (The Caribbean, North and South America, Europe). Traditionally it is used to manage numerous ailments like backache, constipation, cancer, fever in young children, gonorrhea, dysentery, psychosis, hernia, stomach ache, malaria, rheumatism, typhoid etc. It is also used as a cosmetic and food source. The fruit aril is the main constituent of the Jamaican National Dish. This review summarizes its phytochemistry, nutritional properties, ethnobotany and pharmacology. The need to gather up to date information on this plant, to facilitate in vivo and in vitro investigations for verification of some of the local claims necessitated this review. MATERIALS AND METHODS: A literature search was performed on Ackee using ethno botanical text books and scientific databases such as PubMed, Scopus, EBSCO, Science-direct, Google, Google Scholar and other web sources like records from PROTA, PROSEA, JSTOR and Botanical Dermatology Database. RESULTS AND DISCUSSION: This review highlights the traditional uses of parts of the Ackee plant: the bark, the leaves, capsules, roots and seeds. They are used in the management of diverse disease conditions such as diarrhoea, conjunctivitis, fever, internal hemorrhage, dysentery, cutaneous skin infections, and bacterial infections amongst others. Only nineteen compounds have been reportedly isolated from the parts of B. sapida; Alkaloids, quinines, polyphenols, and steroids, their glycosides, sesquiterpenes and triterpenes. Some of the plant extracts and its isolated compounds showed anticancer, antimicrobial, antidiarrheal, antioxidant and hypoglycemic activities both in vitro and in vivo. The seed/leaves have also been used as insect repellants and the leaves have been reported to have lethal effects on larvae of various mosquitoes' species. The oil contains a lot of nutrients and may be considered for edible consumption after safety has been confirmed. Hypoglycin A and the less malignant hypoglycin B are found in the unripe aril of Ackee and consumption results in hypoglycemia, vomiting, gluconeogenesis disruption which can result in coma and death. The untapped economic potential of its fruits is glaring in West Africa countries. CONCLUSION: Though B.sapida has been put to enormous traditional use, the pharmacological studies conducted are not sufficient, most studies are either in-vivo or in-vitro. More work is required (well-designed pharmacological tests, randomized clinical trials) to evaluate these medicinal claims. This review provides a basis for future research. The isolation of more compounds,detailed pharmacological investigations, exploration of food use and detoxification techniques are key areas to investigate.

Quantification of Methylenecyclopropyl Compounds and Acyl Conjugates by UPLC-MS/MS in the Study of the Biochemical Effects of the Ingestion of Canned Ackee (Blighia sapida) and Lychee (Litchi chinensis).

Consumption of ackee (Blighia sapida) and lychee (Litchi chinensis) fruit has led to severe poisoning. Considering their expanded agricultural production, toxicological evaluation has become important. Therefore, the biochemical effects of eating 1 g/kg canned ackee, containing 99.2 µmol/kg hypoglycin A, and 5 g/kg canned lychee, containing 1.3 µmol/kg hypoglycin A, were quantified in a self-experiment. Using ultra-high-performance liquid chromatography/mass spectrometry, hypoglycin A, methylenecyclopropylacetyl-glycine, and methylenecyclopropylformyl-glycine, as well as the respective carnitine conjugates, were found in urine after ingesting ackee. Hypoglycin A and its glycine derivative were also present in urine after eating lychee. Excretion of physiological acyl conjugates was significantly increased in the ackee experiment. Ingestion of ackee led to up to 15.1 nmol/L methylenecyclopropylacetyl-glycine and traces of methylenecyclopropylformyl-carnitine in the serum. These compounds were not found in the serum after eating lychee. Hypoglycin A accumulated in the serum in both experiments.

Saccharides and fructooligosaccharides composition of green and ripe Averrhoa carambola, Blighia sapida and Spondias dulcis fruits.

The maturation of fruits is characterized by numerous compositional changes during ripening and these changes contribute in their quality attributes. This study aimed to assess the contents of saccharides and potential fructooligosaccharides (FOS) of ackee (Blighia sapida Köenig), carambola (Averrhoa carambola) and June plum (Spondias dulcis), at green and ripe stages. Beside glucose and fructose and lower sucrose content, three short chain fructooligosaccharides were identified in ackee fruit, namely 1-kestose (1(F)-ß-d-fructofuranosyl sucrose), nystose (1(F)(1-ß-d-fructofuranosyl)2 sucrose) and DP5 (1(F)(1-ß-d-fructofuranosyl)3 sucrose), while in carambola and June plum DP5 (1(F)(1-ß-d-fructofuranosyl)3 sucrose) was not detected. Ripening stage also affected significantly the contents of these saccharides and sFOS.

Blighia sapida leaves halt elevated blood glucose, dyslipidemia and oxidative stress in alloxan-induced diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Blighia sapida leaves are used in the management of diabetes in Nigeria. Thus the antidiabetic activity of methanolic Blighia sapida leaf extract and its capability to halt oxidative stress and dyslipidemia in alloxan-induced diabetic rats were investigated. METHODS: In vitro antioxidant activity of the extract (0.2-1.0mg/mL) was investigated using 2,2-diphenyl-1-1picrylhydrazyl (DPPH) radical, superoxide ion, hydrogen peroxide, and hydroxyl radical and ferric ion reducing system. Antidiabetic was evaluated in alloxan-induced diabetic rats. RESULTS: The methanolic extract of Blighia sapida leaves at 1.0mg/mL scavenged DPPH, superoxide ion, hydrogen peroxide, and hydroxyl radical at 80.34%, 57.39%, 72.36% and 77.0% respectively, while ferric ion was significantly reduced. Single oral dose of the extract significantly reduced blood glucose level in a dose dependent manner with highest dose producing 18.6% reduction after 240min. Similar reduction was produced after 28 days of extract administration with the highest dose producing 65.65% reduction which compared significantly (P<0.05) with the control group and glibenclamide treated groups. Alloxan-induced diabetic mediated alterations in liver and serum cholesterol, triacylglycerides, high-density lipoprotein cholesterol (HDLc), low density lipoprotein (LDL) cholesterol and very low-density lipoprotein cholesterol (VLDLc) were significantly (P<0.05) restored by the extract. Methanolic extract of Blighia sapida leaves significantly attenuated the decrease in the activities of reactive oxygen species detoxifying enzymes (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glucose 6-phosphate dehydrogenase) in the liver and pancreas of alloxan-induced diabetic rats. Elevation in the concentrations of malondialdehyde, conjugated dienes, lipid hydroperoxides, protein carbonyl, and fragmented DNA was significantly (P<0.05) lowered by Blighia sapida leaves extract. CONCLUSION: Overall, methanolic extract of Blighia sapida leaves at all doses used reduced blood glucose level and prevented oxidative stress and dyslipidemia in alloxan-induced diabetic rats.

Fat profile of Jamaican ackees, oleic acid content and possible health implications / Perfil de grasas del seso vegetal jamaicano, contenido de ácido oleico, y posibles implicaciones para la salud

OBJECTIVE: To re-investigate the composition of ackee oil and unequivocally determine its principal fatty acid components. METHODS: Oil was extracted from the edible portion of ackees harvested in three different studies (I - III) by several analysts; studies I and II utilized composite samples from several trees while study III consisted of ackees from seven separate trees. The oils were either saponified and methylated or trans-methylated and the fatty acid methyl ester content analysed by gas chromatography-mass spectrometry (GC-MS). Relative fatty acid composition was quantified based on chromatographic peak areas while fatty acids were identified by mass spectrometry. The degree of unsaturation of the ackee oils was characterized by determination of the iodine value. RESULTS: Gas chromatography-mass spectrometry data from the three studies were assessed. Relative fatty acid composition for the ackee oils was consistent across the three studies. The major fatty acid components were oleic acid (55.44%), palmitic acid (25.57%) and stearic acid (12.59%); linoleic acid was present in minor to undetectable amounts. An iodine value of 49 was determined which is consistent with the high oleic acid content of the ackee oil. CONCLUSION: The ackee samples analysed were rich in the monounsaturated fatty acid (MUFA) oleic acid.Consideration should be given to potential protective health effects of diets which include ackee.

OBJETIVO: Re-investigar la composición del aceite del seso vegetal (ackee) y determinar de manera inequívoca sus componentes principales de ácidos grasos. MÉTODOS: Se extrajo el aceite de la porción comestible de sesos vegetales de la Blighia sapida (ackee), cosechados como parte de tres estudios diferentes (I - III) por varios analistas. Los estudios I y II utilizaron muestras combinadas de varios árboles de Blighia sapida, en tanto que el estudio III estuvo formado por muestras de siete árboles de Blighia sapida por separado. Los aceites fueron saponificados y metilados o transmetilados, y el contenido de éster metílico de ácidos grasos fue analizado mediante técnicas de cromatografía de gases y espectrometría de masas (GC-MS). La composición relativa de ácidos grasos se cuantificó sobre la base de las áreas cromatográficas pico, mientras que los ácidos grasos se identificaron mediante espectrometría de masas. El grado de insaturación de los aceites de seso vegetal fue caracterizado mediante la determinación del valor de yodo. RESULTADOS: Se evaluaron los datos de la cromatografía de gases acoplada con espectrometría de masas de los tres estudios. La composición relativa de ácido grasos de los aceites de seso vegetal, fue constante a través de los tres estudios. Los componentes principales del ácido graso fueron el ácido oleico (55.44%), el ácido palmítico (25.57%), y el ácido esteárico (12.59%). El ácido linoleico estuvo presente en cantidades que fluctuaron de menores a indetectables. Se determinó un valor de yodo de 49, el cual está en correspondencia con el alto contenido de ácido oleico en el seso vegetal. CONCLUSIÓN: Las muestras analizadas de sesos vegetales de la Blighia sapida (ackee) eran ricas en ácidos grasos monoinsaturados (MUFA por sus siglas en inglés) y ácidos oleicos. Debe prestarse atención a los posibles efectos protectores de la salud de dietas que incluyen sesos vegetales.

Utility of coupled-HSQC experiments in the intact structural elucidation of three complex saponins from Blighia sapida.

The structures of three complex saponins from the fruit pods of Blighia sapida have been elucidated and their (1)H and (13)C NMR spectra assigned employing a variety of one- and two-dimensional NMR techniques without degradative chemistry. The saponins have either four or six monosaccharide units linked to a triterpene aglycone. High-resolution, proton-coupled-HSQC spectra were important for determining both the identities of the intact monosaccharide units and coupling constants in strongly coupled proton spin systems. These NMR experiments will prove crucial as the complexity of saponin structures reaches the limit that can be determined solely by NMR.

Tracking hypoglycins A and B over different maturity stages: implications for detoxification of ackee (Blighia sapida K.D. Koenig) fruits.

Consumption of improperly ripened ackee ( Blighia sapida K.D. Koenig) often results in fatalities. The causal toxin, hypoglycin A, decreases in the edible arilli upon maturity; regulation of hypoglycin A in the arilli is thus critical. Hypoglycin B, also toxic, is confined to the seeds. Hypoglycins A and B were tracked in ackees grown in Jamaica over different maturity stages using RP-HPLC. Studies on the 'Butter' and 'Cheese' ackee varieties and across two different harvest seasons were conducted. In 'Cheese' ackees, hypoglycin A decreased from about 8000 mg/kg in the green arilli and seeds to 271 and 1451 mg/kg, respectively, in the ripe fruit whereas hypoglycin B levels in the seeds increased from 1629 to 11774 mg/kg. The strong inverse relationship demonstrated that hypoglycin B in the seeds serves as a sink for hypoglycin A from the ripening arilli and is thereby involved in the detoxification mechanism of the fruit.

Physicochemical properties of the oil from the fruit of Blighia sapida and toxicological evaluation of the oil-based diet in Wistar rats.

The physicochemical properties of the oil from the fruit of Blighia sapida and the toxicological effect of the oil-based diet on some biochemical parameters of selected rat tissues and serum were studied. The smoke, flash, and fire points as well as peroxide, iodine, and acid values of the fruit oil were significantly lower (P < .05), whereas the specific gravity, relative density, saponification, and ester values compared well with soybean oil. The fruit oil yield was 20.02%. The oil consisted of 22.22% saturated, 56.43% monounsaturated, and 21.35% polyunsaturated fatty acids. It is richer than soybean oil in behenic, palmitoleic, oleic, gadoleic, erucic, and 9,12-eicosanoic acids by 15.70%, 0.89%, 7.22%, 12.05%, 8.27%, and 21.35%, respectively. The liver- and kidney-body weight ratios as well as the serum concentrations of cholesterol and high-density lipoprotein cholesterol of the rats maintained on diet formulated with the oil from the fruit of B. sapida increased significantly (P < .05), but the triglyceride and atherogenic index decreased (P < .05). The low-density lipoprotein cholesterol concentration and the heart-body weight ratio of the rats fed with the fruit oil diet compared well (P > .05) with those on soybean oil-based diet. Animals fed with the fruit oil-based diet had their activities of liver glutamate oxaloacetate transaminase and glutamate pyruvate transaminase as well as alkaline phosphatase activities of the liver and kidney decreased with corresponding increase in the serum enzymes. These results suggest that oil from B. sapida fruit could be edible and may be explored as raw materials in the paint, margarine, and soap industries. The oil is also unlikely to predispose the animals to cardiovascular risk, but may labilize the plasma membrane of the hepatocytes and nephrons. It may also have a negative effect on the metabolism and regulation of amino acid in the animals. Therefore, the oil from B. sapida fruit may not be completely safe for consumption.

Structural characterization of hypoglycin B, a diastereomeric dipeptide from the ackee fruit (Blighia sapida Koenig) by NMR experiments.

The dipeptide hypoglycin B, one of two toxins of the ackee fruit (Blighia sapida Koenig), was characterized for the first time by NMR spectral data, which led to the discovery that it exists naturally as a pair of diastereomers. No distinction was observed in the (1)H NMR signals of the diastereomers; however, complete and distinct (13)C NMR assignments for the individual diastereomers were made. The (13)C NMR spectrum of hypoglycin B compared very well with that of the corresponding signals in the spectrum for hypoglycin A, which is one of its constituent amino acids. The (1)H and (13)C NMR assignments were further supported by DEPT, gCOSY, gHSQC and gHMBC experiments.

Characterisation of Blighia sapida (Sapindaceae) seed oil and defatted cake from Benin.

A sample of Blighia sapida seeds collected in Benin has been analysed and the results are compared to the scarcely available literature data. The chemical analysis of seed oil shows a saponification value of 145 and an iodine value of 66, consistent with the high mono-unsaturated fatty acids (FAs) content (63.8 wt%). The most interesting feature is the prominent concentration of eicosenoic acid (48.4 wt%). Arachidic acid being the main component within the saturated group, the C20 FAs fraction accounts for 68.4 wt%, thus making the peculiar composition of this oil. Among the unsaponifiable fraction (2.4 wt%), the major sterol is stigmasterol (54.6 wt%), surprisingly over passing beta-sitosterol. Tocols (338 ppm) contains mainly alpha- and gamma-tocopherol. Regarding the defatted cake, results show the prominent position of starch and a noticeable amount of proteins and fibers (44.2, 22.4, 15.6 wt%, respectively). Seventeen amino acids were identified together with valuable minerals (total ashes 3.5 wt%). Possible uses of oil and defatted cake are discussed.

Ackee (Blighia sapida) hypoglycin A toxicity: dose response assessment in laboratory rats.

Hypoglycin A, the toxin found in the ackee fruit, has been reported in the literature as the causative agent in incidences of acute toxicity termed Jamaican vomiting sickness or toxic hypoglycemic syndrome. Hypoglycin A toxicity in this study was determined by feeding male and female Sprague-Dawley rats a control diet and ackee diets that contained 4-3840 ppm of hypoglycin. The fixed dose method was used to quantify the acute toxic dose of hypoglycin A and was determined by feeding a diet consisting of the lowest hypoglycin A concentration; this was increased to the next highest dose after 24h until toxicity was observed. The maximum tolerated dose (MTD) of hypoglycin A was determined by feeding rats the ackee and control diets over a 30-day period. The acute toxic dose for male and female rats was 231.19+/-62.5 5mg hypoglycinA/kgBW and 215.99+/-63.33 mg hypoglycinA/kgBW, respectively. This was considerably greater than the dose of 100 mg hypoglycin/kgBW reported in a previous study when aqueous hypoglycin was administered orally. The MTD of hypoglycin A in both male and female rats was 1.50+/-0.07 mg hypoglycinA/kgBW/day. These findings suggest that the form in which hypoglycin in ackee is administered could affect the toxicological properties it exhibits. Therefore, for the purpose of a hazard assessment, it may be best administered within the matrix of the fruit, which is the form that humans consume it.

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.