Phenolic acids in foods: an overview of analytical methodology

Phenolic acids are aromatic secondary plant metabolites, widely spread throughout the plant kingdom. Existing analytical methods for phenolic acids originated from interest in their biological roles as secondary metabolites and from their roles in food quality and their organoleptic properties. Recent interest in phenolic acids stems from their potential protective role, through ingestion of fruits and vegetables, against oxidative damage diseases (coronary heart disease, stroke, and cancers). High performance liquid chromatography (HPLC) as well as gas chromatography (GC) are the two separation techniques reviewed. Extraction from plant matrixes and cleavage reactions through hydrolysis (acidic, basic, and enzymatic) are discussed as are the derivatization reagents used in sample preparation for GC. Detection systems discussed include UV-Vis spectroscopy, mass spectrometry, electrochemical, and fluorometric detection. The most common tandem techniques are HPLC/UV and GC/MS, yet LC/MS is becoming more common. The masses and MS fragmentation patterns of phenolic acids are discussed and tabulated as are the UV absorption maxima.     

Determination of hydrolyzable tannins (gallotannins and ellagitannins) after reaction with potassium iodate

A widely used method for analyzing hydrolyzable tannins afer reaction with KIO(3) has been modified to include a methanolysis step followed by oxidation with KIO(3). In the new method, hydrolyzable tannins (gallotannins and ellagitannins) are reacted at 85 degrees C for 20 h in methanol/sulfuric acid to quantitatively release methyl gallate. Dried plant samples can be methanolyzed under the same conditions to convert hydrolyzable tannins to methyl gallate. Oxidation of the methyl gallate by KIO(3) at pH 5.5, 30 degrees C, forms a chromophore with lambda(max) 525 nm, which is determined spectrophotometrically. The detection limit of the method is 1.5 microg of methyl gallate, and with plant samples, relative standard deviations of less than 3% were obtained.     

Interaction of flavonoids with bovine serum albumin: a fluorescence quenching study

The interaction between four flavonoids (catechin, epicatechin, rutin, and quercetin) and bovine serum albumin (BSA) was investigated using tryptophan fluorescence quenching. Quenching constants were determined using the Stern-Volmer equation to provide a measure of the binding affinity between the flavonoids and BSA. The binding affinity was strongest for quercetin and ranked in the order quercetin > rutin > epicatechin = catechin. The pH in the range of 5-7.4 does not affect significantly (p < 0.05) the association of rutin, epicatechin, and catechin with BSA, but quercetin exhibited a stronger affinity at pH 7.4 than at lower pH (p < 0.05). Quercetin has a total quenching effect on BSA tryptophan fluorescence at a molar ratio of 10:1 and rutin at approximately 25:1. However, epicatechin and catechin did not fully quench tryptophan fluorescence over the concentration range studied. Furthermore, the data suggested that the association between flavonoids and BSA did not change molecular conformation of BSA and that hydrogen bonding, ionic, and hydrophobic interaction are equally important driving forces for protein-flavonoid association.     

Morphometric and steroid hormone changes associated with experimental anovulatory follicles in the sow.

Steroid levels and ovarian follicular morphology were examined in sows on days 19 and 26 (day 5 of next cycle) after injection of adrenocorticotropic hormone (ACTH) or dexamethasone (DXM). Five sows received DXM (30 micrograms/kg bodyweight, intramuscularly) at 12 h intervals from days 9 to 14. Another five sows were given ACTH (2 IU/kg bodyweight, intramuscularly) from day 17 to day 19 or the end of estrus. Five control sows received no treatment. Ovulation occurred only in control sows and progesterone was significantly elevated at day 26. Estradiol values in ovarian vein blood were low but variable on day 19 in DXM- and ACTH-treated animals. Androstenedione values were lower (p less than 0.05) on both days in sows receiving DXM but not in those given ACTH compared to control values on day 19. Morphometric analysis, based on six follicles in each of three sows from each treatment group, indicated that follicular and antral diameters and granulosa cell numbers did not differ for either hormone treatment group on either day compared to those of control sows on day 19. The mitotic index suggested that cell replication continued. However, pyknotic and karyorrhectic nuclei were also seen in the hormone treatment groups. Follicles and oocytes from both DXM- and ACTH-treated sows showed signs of early degenerative changes including disorganization of cumulus cells and large lipid droplets in the cytoplasm of oocytes. Significant differences from control follicles in granulosa cell density and theca interna cell density suggested an association with the altered steroid hormone secretion.     

BLUETONGUE CERTIFICATION—AUSTRALIAN POLICY

Australian quarantine policies with respect to BT are based on regarding this disease as one of high risk and major potential economic importance to our ruminant population. There are deficiencies in our knowledge of world distribution, epidemiology and pathogenesis. There may be unknown vectors and unsuspected animal reservoir hosts. The international distribution of BT could be extending through the movements of insects or cattle. If introduced into Australia, the cattle and sheep populations would probably be at continuous risk as eradication would be difficult or impossible. Costs to the sheep industry at least could be high and productivity gravely affected. To reduce the probability of introduction, imports of ruminants and semen have, for the last 16 years, been restricted to a very small number of countries presumed free from BT. Future policies will almost certainly be based on these same considerations taking into account scientific advances in diagnosis and virus detection. The establishment of an off-shore high security quarantine station will facilitate imports of ruminants even from known BT-infected countries.     

Responses of Juvenile Golden Shiner Notemigonus crysoleucas Fed Semipurified or Practical Diets With or Without Supplemental Ascorbic Acid

A 12-wk feeding trial was conducted in a flow-through system to determine whether juvenile golden shiner Notemigonus crysoleucas have a dietary requirement for ascorbic acid. Triplicate groups of 30 fish each weighing 0.44 g initially were fed semi-purified (casein) diets supplemented with 0 or 250 mg ascorbic acid/kg, or practical (fish meal) diets supplemented with 0 or 250 mg ascorbic acid/kg. The form of ascorbic acid used was L-ascorbyl-2-polyphosphate. Survival was higher in fish fed the semi-purified diet with 250 mg ascorbic acid/kg than in those fed the unsupplemented semi-purified diet, while weight gain did not differ between treatments. Weight gain was higher in fish fed the practical (fish meal) diet supplemented with 250 mg ascorbic acid/kg than in those fed the unsupplemented practical diet, while survival did not differ between treatments. No other signs of ascorbic acid deficiency were observed, but fish fed the casein diets had skin and fin erosion indicative of a possible bacterial infection. Whole-body concentrations of ascorbic acid (total, reduced, and percent of reduced ascorbic acid) were higher in fish fed semi-purified or practical diets supplemented with 250 mg ascorbic acid/kg than in fish fed unsupplemented diets. Golden shiners appear to require a dietary source of ascorbic acid for growth or survival, depending on experimental conditions. The two main differences in the study were in diet composition (semi-purified versus practical diets) and apparent health status of fish fed the different diet types. Additional studies are needed to define requirements further.