A rapid method for the quantification of steroidal glycoalkaloids by reversed phase HPLC

Summary Methods were developed for clean-up of potato tuber extracts on solid phase extraction CN-cartridges and for analysis of steroidal glycoalkaloids by reversed phase HPLC. The alkaloids α-solanine, α-chaconine, α-solasonine and α-solamargine could be separated on C₈ or C₁₈ reversed phase columns using a mobile phase of acetonitrile/Tris-buffer (3∶2, v/v). The analysis time of tuber extracts containing these alkaloids was less then 4 min, if the pH of the mobile phase was adjusted to 7.4 and run at a flow rate of 0.5 ml/min.     

Einfluss von Lagerung und Sorte auf die Verteilung der Glykoalkaloide in der Kartoffelknolle

Zusammenfassung Fünf Speisekartoffelsorten wurden w?hrend einer sechsmonatigen Lagerung fünfmal auf α-Solanin und α-Chaconin untersucht. Es wurden vier Gewebebereiche der Knolle getrennt überprüft. Der Gesamtglykoalkaloidgehalt betrug im Augenbereich 164, in der restlichen Schale 101 und in der Gef?ssbündelzone mit ?usserem Speichergewebe 8mg/100g. Trockensubstanz. Im restlichen Innenk?rper waren nur noch Spuren (0.164mg/100g TS) nachzuweisen. Zwischen den Sorten und diesen Gewebepartien traten statistisch gesicherte Wechselwirkungen auf, die im wesentlichen von einer Sorte verursacht wurden. W?hrend der Lagerung sind die Alkaloidgehalte leicht, aber statistisch nicht gesichert gesunken. α-Solanin und α-Chaconin kamen generell im Verh?ltnis von etwa 1∶1.5vor; die Korrelation zwischen beiden Formen ist sehr eng (r=0.865). Bezogen auf die gesamte Knolle lagen die Gesamtglykoalkaloide je nach Sorte zwischen 2.4 und 8mg/100g Frischsubstanz und damit im gesundheitlich unbedenklichen Bereich.

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Summary Five table potato cultivars (Erna, Granola, Gusto, Ulse, Margit) from the same source were examined for their content of α-solanine and α-chaconine during a sixmonths storage period. For the testing and subsequent analysis the tubers were dissected into four kinds of tissues: eye-zone, peel, vascular bundle region, medulla. The total glycoalkaloid content (TGA) in the total tuber mass varied between 2.4 and 8mg per 100g fresh matter depending on cultivar (Fig. 1). Despite significant differences between cultivars (Table 1), all potato cultivars remained within the safe range during the entire storage period. Remarkable differences in TGA per 100g dry matter could be noted in the tuber parts. For the eye-zones a TGA of 164mg was found; the peel region showed 101mg and the vascular bundle zone 8mg. The medulla of the tubers contained only traces of TGA (0.164mg/100g dry matter, Fig. 2). The statistical analysis only revealed significant interactions between cultivars and tuber parts (Table 1), mainly caused by the cultivar Gusto which showed higher contents of α-solanine and α-chaconine in its vascular bundle regions (Figs 3–5). The ratio between amounts of α-solanine and α-chaconine was approximately 1∶1.5, and after eliminating effects of cultivar, storage and tuber parts, the correlation coefficient between amounts was 0.865. This close correlation can be used for breeding. The non-significant, slightly negative effect of the six-months storage period on alkaloid content was noteworthy. It is assumed that only after long storage periods or through inappropriate storage (sprouting of tubers) a significantly higher glycoalkaloid content may be expected.

     

Combination of On-Line Solid-Phase Extraction with LC-MS for the Determination of Potentially Hazardous Glycoalkaloids in Potato Products

A simple and rapid method for the determination of naturally occurring, potentially hazardous glycoalkaloids (GAs) in potatoes and their products has been developed. The procedure is based on the on-line solid-phase extraction of the acetic acid extracts from potato products and combined with liquid chromatography (LC)-mass spectrometry (MS) in a fully automated system (Symbiosis™, Spark Holland Instruments, Emmen, The Netherlands). As sorbent material HySphere™ 18HD was used for alkaloid enrichment. GAs were eluted with the LC gradient and directly analysed by MS. Detection of the analytes was achieved in the sensitive multiple reaction monitoring mode using two characteristic ions (m/z 98 as a qualifier for GAs and m/z 868.3 as a quantifier for α-solanine or m/z 852.4 for α-chaconine). Typical validation data for method precision (v _k α-solanine = 5.3–6.5, v _k α-chaconine = 3.4–15.4), accuracy (average recovery of α-solanine = 84%, average recovery of α-chaconine = 87%) and linearity over the range from 1 to 1,000 ng ml⁻¹ (R ² = 0.9915 for α-solanine, R ² = 0.9939 for α-chaconine) with detection limits of 0.3 ng ml⁻¹ for α-chaconine and 0.5 ng ml⁻¹ for α-solanine were obtained. GA contents of commercial potato products were determined by the new on-line method and afterwards compared with those obtained with an established high-performance LC routine procedure. Better performance of the on-line procedure was obvious from the standard deviations of both methods. Other advantages included a strong reduction of overall analysis time, human intervention and solvent consumption as well as waste production. The time required for the on-line analysis was 5 min, which would allow processing of almost 100 samples in 8 h.     

Identification and Quantification of Steroidal Alkaloids from Wild Tuber-Bearing <Emphasis Type="Italic">Solanum</Emphasis> Species by HPLC and LC-ESI-MS

Wild Solanum species are characterized by several types of glycoalkaloids (GAs), which are usually not found in commercial potato cultivars. These alkaloids serve as defence compounds against herbivores and microorganisms, especially fungi. In this study, the GA composition of tuber and leaf material of 17 wild Solanum species was analysed qualitatively by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and quantitatively by high-performance liquid chromatography. The GAs identified were α-solanine, α-chaconine, β-chaconine, solasonine, solamargine, demissine, dehydrodemissine, α-tomatine, dehydrotomatine, commersonine, dehydrocommersonine, leptine I and leptine II. Most species accumulated the common potato GAs α-solanine and α-chaconine in tubers and solasonine and solamargine additionally in leaves. In some species, such as S. acaule ssp. acaule, S. demissum and S. polyadenium, substantial amounts of unusual alkaloids were detected. By using LC-ESI-MS, we detected several minor alkaloids such as dehydrogenated forms of α-tomatine, demissine and commersonine for the first time. Total GA content, expressed as the sum of the four main alkaloids α-solanine, α-chaconine, solamargine and solasonine differed from species to species. In general, GA contents in leaf tissue were higher and GA patterns were more complex than those of tubers.     

Glycoalkaloid accumulation during tuber development of early potato cultivars

Summary Plants of early potato cultivars were sampled at successive harvests during tuber development and individual tubers were analysed for α-solanine and α-chaconine using high-performance liquid chromatography. Mean tuber total glycoalkaloid concentration (α-solanine plus α-chaconine) per plant decreased with time and statistically significant differences between cultivars were observed in the patterns of decline during tuber growth. The mean tuber ratio of α-chaconine to α-solanine increased during tuber development and was also affected by genotype. Total glycoalkaloid concentration for individual tubers of marketable sizes (up to 50 g fresh weight) exceeded the safety limit of 20 mg per 100 g fresh weight for cultivars Home Guard and British Queen but not for Rocket. These differences were attributable to differences in both rates and patterns of glycoalkaloid accumulation during tuber development, although in all cultivars results were consistent with glycoalkaloid accumulation occurring in the expanding peripheral cell layers of tubers for a considerable period after tuber initiation.     

Determination of the glycoalkaloids α-solanine and α-chaconine levels in 18 varieties of potato (Solarium tuberosum L.) grown in Oman

Summary The glycoalkaloid content in 18 potato varieties grown in Oman was measured by HPLC using peel and flesh tissues. The total glycoalkaloid content of the tubers was found to be variety dependent and varied greatly between the 18 varieties. The total glycoalkaloid in the tubers of seven varieties was within the safe limit of ≤200 mg kg^{− 1} FW recommended for food safety, whereas the remaining 11 varieties exceeded this upper limit. The principal glycoalkaloids α-chaconine and σ-solanine were present in higher concentration in the peel than in the flesh of all varieties with the former being the principal alkaloid in the peel. The flesh of all of the 18 potato varieties contained total glycoalkaloid levels lower than the upper safety limit. The leaves of the 18 potato varieties were also analyzed for glycoalkaloids. The levels of glycoalkaloids in expanding leaves were higher than that of expanded leaves with the exception of six varieties. The principal glycoalkaloid in expanding and expanded leaves was α-chaconine. No correlation was observed between the glycoalkaloids levels of in the leaves and tubers.     

Inheritance study of tuber glycoalkaloids in a wild potato,Solanum chacoense bitter

Studies were made of segregating families from crosses between accessions of Solanumchacoense Bitter differing in steroid glycoalkaloids. The major glycoalkaloids were isolated from tubers and identified by thin-layer chromatography. Simple segregation ratios of 3:1 in an F₂ and 1:1 in backcrosses were obtained for the presence:absence of solanine, chaconine, and commersonine. To explain the simultaneous segregation of the three glycoalkaloids, we tentatively hypothesize that: (1) solanine and commersonine are determined by alternative codominant alleles at a single gene locus; (2) a major gene for chaconine segregates independently of the solanine/commersonine locus; and (3) a recessive gene linked with the commersonine allele is epistatic to the chaconine gene, causing the production of β-chaconine instead of the usual α-chaconine. The F₂ distribution could be fit to the ratio 3:6:3:1:2:1, seen as combining the 1:2:1 segregation of solanine: solanine + commersonine: commersonine and the 3:1 segregation of presence: absence of chaconine.     

Inhibition of acetyl cholinesterase by solanaceous glycoalkaloids and alkaloids

Seven solanaceous glycoalkaloids (α-chaconine, β₂-chaconine, α-solanine, dehydrocommersonine, commersonine, demissine and tomatine) and three alkaloids (solanidine, tomatidine and demissidine) were tested for their ability to inhibit acetyl cholinesterase in anin vitro system. Glycoalkaloids at concentrations of 33–41 parts per million (ppm) gave cholinesterase inhibition ranging from 4.2 to 26.8%. All three alkaloids had lower anticholinesterase (4.2 to 15.4%) than the seven glycoalkaloids, except for tomatine. Two well-known cholinesterase inhibitors, carbaryl, a carbamate insecticide and Guthion, an organophosphate insecticide, were also tested for comparison. It required only 1 to 2 ppm of the insecticides to give the same average percent inhibition as was observed with the glycoalkaloids at concentrations of 33–35 ppm.     

A rapid quantitative determination of the individual glycoalkaloids in tubers and leaves ofSolanum tuberosum L.

Summary A rapid quantitative determination of the individual glycoalkaloids solanine and chaconine is described. The glycoalkaloids were extracted by blending with methanol and the dry residues obtained were worked up by extraction withn-butanol. After separation of them by thin layer chromatography, they were assayed by in situ fluorodensitometry of the chromatograms. The recoveries levels obtained by the newly developed extraction and work-up procedure were similar to those obtained by the method of Sachse and Bachmann. Potato tubers and leaves, and industrial potato protein were analysed with this method and its precision was equal or better than that of other methods. It can also be used as a semiquantitative thin layer chromatography assay of potato glycoalkaloids.

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Zusammenfassung Beschrieben wird eine schnelle und quantitative Bestimmung der Glykoalkaloide Solanin und Chaconin. Die Prim?r-Extraktion der Glykoalkaloide durch Eluieren mit Methanol wurde mit der nach der Methode von Sachse und Bachmann durchgeführten Extraktion unter Rückfluss verglichen. Nach dem Aufarbeiten der Extrakte durch Ausf?llung wurden die Glykoalkaloide dünnschichtchromatografisch (DC) getrennt und die Chromatograpmme durch in situ Fluorodensitometrie gemessen. Da sich keine signifikanten Unterschiede (α=0,05) zwischen den beiden Extraktionsmethoden ergaben (Tabelle 1) wurde die weniger arbeitsintensive Methode des Eluierens für die Extraktion der anderen Proben gew?hlt. Um die zeitaufwendige Methode des Ausf?llens zu umgehen, prüften wir für die Aufarbeitung zwei Modifikationen der von Crabbe & Fryer (1980) benutztenn-Butanol-Extraktion. Der trockene Rückstand der Prim?rextraktion wurde in einer Mischung gleicher Teile von Butanol, ges?ttigt mit einprozentigem Ammoniak und 1% Ammoniak ges?ttigt mitn-Butanol aufgenommen. Nach der Trennung der Schichten wurde die Butanolschicht für die quantitative DC verwendet. Bei der zweiten Modifikation wurde der trockene Rückstand in verdünnter Essigs?ure gel?st und zentrifugiert. Die klare saure L?sung wurde alkalisch gemacht und mitn-Butanol ausgechüttelt. Da in diesem Fall die Verwendung ges?ttigter L?sungsmittel nicht m?glich war, wurde ein Korrekturfaktor für die Volumenver?nderung dern-Butanol-Schicht bestimmt. Nach dem Schütteln blieben von den ursprünglich 10 mln-Butanol 9,5 ml übrig. Die Ergebnisse dieser beiden Aufarbeitungsmethoden wurden mit den durch Ausf?llung erhaltenen Ergebnissen verglichen (Tabelle 2). Bei der einfachenn-Butanol-Extraktion war die Trennung der Glykoalkaloide durch DC bei Proben mit einem geringen Glykoalkaloidgehalt insofern gest?rt als sich zu hohe Glykoalkaloidwerte ergaben. Die Extraktion mit Essigs?ure/n-Butanol ergab Werte, die gleich hoch oder geringfügig h?her waren, als nach der Ausf?llung. Die DC-Trennung der Glykoalkaloide wurde nicht durch andere fluoreszierende Substanzen gest?rt. Die Wiederfindungsrate der gesamten Methode: Eluieren und Essigs?ure/n-Butanol-Extraktion entsprach der Methode von Sachse & Bachmann (1969) und zeigte eine nahezu vollst?ndige Extraktion der Glykoalkaloide an (Tabelle 3). Die Methode ist genau und empfindlich wie Tabelle 4 für die Glykoalkaloidwerte mehrerer Proben zeigt. Eine ann?hernde Bestimmung des Glykoalkaloidgehaltes kann durch einen visuellen Vergleich von Proben- und Standardflecken durchgeführt werden. Für diese halbquantitative DC genügt die einfachen-Butanol-Extraktion.

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Résumé Une méthode de détermination quantitative rapide de la solanine et de la chaconine est décrite. La première extraction des glucoalcalo?des par mélange avec le méthanol a été comparée avec l'extraction sous réfrigérant à reflux, selon la méthode de Sachse et Bachmann. Après le traitement des extraits par précipitation, les glucoalcalo?des ont été séparés par chromathographie sur couche mince (TLC) et évalué par fluorodensitométrie directement sur les chromatogrammes. Dans la mesure où aucune différence significative (α=0,05) n'a été trouvée entre les deux méthodes d'extraction (tableau 1), celle demandant le moins de travail a été choisie pour l'extraction de ces substances dans les autres échantillons. Pour gagner du temps sur la méthode de précipitation, nous avons procédé à deux modifications au niveau de l'extraction aun. butanol utilisée par Crabbe & Fryer (1980). La première consistait à dissoudre la résidu sec de l'extraction précédente dans un mélange, en parties égales, den. butanol saturé avec 1% d'ammoniaque et de 1% d'ammoniaque saturé den. butanol. Après séparation des deux phases, la phasen. butanol a été utilisée pour l'essai quantitatif en chromatographie sur couche mince. La seconde modification consistait à dissoudre la résidu sec dans de l'acide acétique dilué et à centrifuger la solution acide. Cette solution clarifiée était alcalisée et l'extraction était faite aun.butanol; l'utilisation d'un solvant saturé étant impossible dans ce cas, un facteur de correction de volume a été déterminé pour len.butanol. Après agitation on conservait 9,5 ml des 10 ml d'origine de la phasen.butanol. Les résultats de ces deux procédés ont été comparés avec ceux obtenus par la méthode de précipitation (tableau 2). Avec la seule extraction aun.butanol la séparation des glucoalcalo?des par chromatographie a été perturbée pour les échantillons ayant une faible teneur en donnant des valeurs trop élevées. L'extraction à l'acide acétique/n.butanol a donné des valeurs égales ou légèrement supérieures à celles de la méthode de précipitation. La séparation des glucoalcalo?des n'a pas été perturbée par les autres composés fluorescents. Les valeurs obtenues par la méthode totale=mélange avec méthanol et extraction à l'acide acétique/n.butanol ont été approximativement les mêmes que celles obtenues par la méthode de Sachse & Bachmann (1969); elles indiquent une presque complète extraction des glucoalcalo?des (tableau 3). La méthode est précise et sensible comme le montrent les valeurs indiquées dans le tableau 4. Une évaluation approximative de la teneur en glucoalcalo?des peut être obtenue par comparaison visuelle entre l'échantillon et des bandes témoins standard. Pour cette chromatographie semi-quantitative la simple extraction aun.butanol est suffisante.

     

Evaluation of Vitamins and Glycoalkaloids in Potato Genotypes Grown Under Organic and Conventional Farming Systems

Potato tubers provide people not only with carbohydrates but also with other essential compounds for human health. Several investigations have compared the quality of crops grown under organic and conventional farming systems. The aim of the present study was to evaluate the concentrations of vitamins C, B₁ and B₂ as well as glycoalkaloids (α-solanine and α-chaconine) in potato tubers of different genotypes and to estimate the farming system (conventional and organic) impact on the content of these compounds. Twenty potato genotypes were grown under organic and conventional farming systems in Priekuli (Latvia) during 2010 and 2011. The concentrations of vitamins C, B₁ and B₂ as well as glycoalkaloids (α-solanine and α-chaconine) were significantly influenced by the potato genotype. The vitamin B₁ concentration of potato genotypes grown in the organic field exceeded that of potato genotypes grown in the conventional field. A significant negative correlation was found between the vitamin C concentration and tuber yield only in the conventional field. No significant correlations were found between the other potato characteristics, such as starch content, tuber taste after boiling and glycoalkaloids, vitamins C, B₁ and B₂. The high broad-sense heritability for all tested traits (vitamin C 93%, vitamin B₁ 92%, α-solanine 88%, α-chaconine 84% and vitamin B₂ 70%) demonstrated that the high genetic diversity in the potato genotype population accounted for the phenotypic expression. The present study suggested that the breeding of new potato varieties with improved nutritional quality can be launched, and that the varieties that will be developed can be produced in an environmentally friendly way.     

Amylolytic activity in stored potato tubers. 1. Estimation usingp-nitrophenyloligosaccharides

Summary Methods for the estimation of amylolytic activity are reviewed. A procedure for the routine extraction of amylolytic activity from freeze-dried powder prepared from potato tubers is described. The extraction medium is buffered at pH 7.0 and contains glycerol, dithiothreitol, calcium chloride and the non-ionic detergent, nonidet P-40. α-Amylase activity and exoamylolytic activity were estimated in crude extracts of potato tubers using the substrates, blockedp-nitrophenyl-maltoheptaoside andp-nitrophenyl-maltopentaoside respectively. These substrates are included in kits supplied by Biocon, (UK) Ltd to measure the α- and ?-amylase activity in cereals. The validity of using these kits for the determination of α- and ?-amylase activity in potato tuber tissue is discussed.     

Characteristics of two major lectins from mungbean (Vigna radiata) seeds

Two major lectins, MBL-I and MBL-II, were purified fromVigna radiata L. seeds using ion-exchange and gel filtration chromatography techniques. MBL-I was found to be a tetramer with native M.W. of 132 kDa and subunit M.W. of 33 kDa having α-galactosidase activity. MBL-II consisted of two monomeric lectins with M.W. of 94 kDa and 89 kDa which were associated mainly with β-galactosidase activity. Both MBL-I and MBL-II are D-galactose-specific lectins.     

Amylolytic activity in stored potato tubers. 2. The effect of low-temperature storage on the activities of α-and β-amylase and α-glucosidase in potato tubers

Summary Tubers of the potato cultivars Record, Wilja, Pentland Dell and Brodick (formerly clone 137371) were sampled before and after storage at either 4°C or 10°C. Reducing sugar content stayed constant during storage at 10°C in all four cultivars but rose greatly during the first 6–12 weeks of storage at 4°C in Record, Wilja and Pentland Dell but not in Brodick. Amylolytic activity was determined after 5 weeks storage using blockedp-nitrophenyl maltoheptaoside as substrate for α-amylase,p-nitrophenyl maltopentaoside as substrate for β-amylase, andp-nitrophenylglucopyranoside as substrate for α-glucosidase. The values obtained from tubers stored at 4°C were higher than those from tubers stored at 10°C, the differences being much less in Brodick than in the other three cultivars.     

Changes in the content of L-ascorbic acid,glucose, fructose,sucrose and total glycoalkaloids in potatoes (cv. Bintje) stored at 7, 16 and 28°C

Summary We investigated the changes in the content of L-ascorbic acid (L-AA), glucose, fructose, surose and total glycoalkaloids (TGA) during storage of potatoes, cv. Bintje, for 12 weeks at 7,16 and 28°C. The initial amount of L-AA was 8.2 mg/100g fresh weight; after 12 weeks' storage at 7°C it had decreased, while at 16 and 28°C it had increased. The sugar content went up most in potatoes stored at 28°C, but remained below the level at which potatoes taste sweet. The sugar content rose slightly at 16°C, while it decreased in tubers stored at 7°C. The initial TGA content, in equivalent amounts of α-solanine, was 3.6 mg/100 g fresh weight; after 12 weeks' storage it had decreased slightly at all three storage temperatures.     

The Consumption of <Emphasis Type="Italic">Hibiscus sabdariffa</Emphasis> Dried Calyx Ethanolic Extract Reduced Lipid Profile in Rats

The scientific basis for the statement that plants and their active constituents play an important role in the prevention of chronic and degenerative diseases is continously advancing. The object of the present study was to evaluate the effect of Hibiscus sabdariffa L. dried calyx ethanolic extract on the serum lipid profile of Sprague-Dawley rats. The rats were fed during 4 weeks with either a basal diet, containing high cholesterol (1%), cholic acid (0.25%), lard oil (10%), or a supplemental diet with H. sabdariffa extract at 5%, 10%, and 15% levels (SD₅, SD₁₀, SD₁₅). Weight gain and faeces dry weight were both very significantly less (p ≤ 0.01) in SD₁₀ and SD₁₅ groups as compared to the control group, as was food efficiency in SD₁₅. In the case of SD₁₀, food efficiency was only significantly lower (p ≤ 0.05). Faeces dry weight in SD₅ however, was the only one found to be significantly higher (p ≤ 0.05). Triacylglycerols and LDL levels were both significantly less (p ≤ 0.05) in all groups (SD₅, SD₁₀, and SD₁₅) as compared to the control. For total lipids, SD₁₀ and SD₁₅ showed significantly lower levels (p ≤ 0.05), whereas very significant differences (p ≤ 0.01) were observed in the case of SD₅. All groups had lower cholesterol levels compared to controls; however, only the SD₅ group was statistically significant (p ≤ 0.05). No significant results were found in any group in the cases of either phospholipid or HDL levels. A hypothesis of hibiscus acid racemization, (+)-HCA to (−)-HCA, mediated by intestinal flora enzymes possibly explains the significant triacylglycerol decrease in all experimental groups. VLDL, the precursor of LDL, is composed predominantly of triacylglycerols, suggesting that the significant decrease in LDL is related to observed triacylglycerol synthesis inhibition. Five percent H. sabdariffa extract addition showed the best results in the reduction of serum lipids under study conditions.     

The extraction and partial purification of a rhamnosidase from Colorado potato beetle larvae (Leptinotarsa decemlineata (Say))

An enzyme which hydrolyzed a terminal rhamnose moiety from α-chaconine was extracted and partially purified from Colorado potato beetle (CPB) (Leptinotarsa decemlineata) larvae. This ammonium sulfate precipitated enzyme fraction resulted in nearly 97% metabolism of α-chaconine after a 144 hour incubation period at 42 C as determined by high performance liquid Chromatographic (HPLC) analysis. Approximately 81% of the metabolized α-chaconine was detected asβ ₂-chaconine. The isolated enzyme fraction from CPB larvae exhibited no metabolism of α-solanine. Metabolism was also followed over time by thin layer chromatography (tlc).     

Isolaton,partial purification and characterization of a potato peel glycoalkaloid glycosidase

The enzymes involved in the hydrolysis of the rhamnose moieties fromα-chaconine were isolated and partially purified by ammonium sulfate precipitation from the peels of Kennebec and Wauseon potatoes. Enzyme activity was found to be present in the 60 to 80% ammonium sulfate precipitate. Enzyme activity was increased by the use of non-ionic detergents with a mixture of Triton X-100:Triton CF-54 providing the greatest increase in activity. The pH and temperature optimums for the enzymes from both varieties were found to be 6.0 and 42 C, respectively. The route ofα-chaconine metabolism (either through B₁-or B₂-chaconine) was dependent upon incubation temperature. The effect of substrate concentration on enzyme activity was determined for both varieties and the Kms calculated from Lineweaver-Burke double reciprocal plots. The Km values for the Kennebec and Wauseon rhamnosidases were found to be 9.2×10^?4 and 1.7×10^?5M, respectively.     

Carbon dioxide and methane gas permeations in thermally annealed and chemically cross-linked commercial polyimide hollow fiber membrane

Two types of modifications were performed on a commercial polyimide (PI) hollow fiber membrane for carbon dioxide (CO₂) and methane (CH₄) gas permeations. Thermal annealing was conducted between 50- and 200 °C while chemical cross-linking was performed using 0.1- to 1.0 wt% of N, (1-Naphthyl) ethylene-diamine dihydrochloride (NED). Membrane characterization revealed densification of the thermally annealed PIs. But formation of macrovoids was observed in PIs annealed near its glass transition temperature (207 °C). Fourier transform infrared spectroscopy confirmed the successful cross-linking of NED with PI. Highest CO₂ permeance was obtained from pristine PI (P/L=225 GPU) but it also had the lowest selectivity (α=72). The performances of thermally annealed (P/L=160–219 GPU, α=76–106), NED crosslinked (P/L=68–139 GPU, α=65–95) and thermally induced NED cross-linked (P/L=51-91 GPU, α=98–138) PIs varied according to modification conditions. Among the modified membranes, highest CO₂ permeance was obtained from thermally annealed PI at 100 °C (P/L=211GPU, α=106) while thermally induced NED cross-linked PI (100 °C, 0.5 wt% NED) exhibited the highest selectivity (P/L=91, α=138). Both modified membranes are the best candidates for CO₂/CH₄ separation.     

Antidiabetic and Antioxidant Effects of Polyphenols in Brown Alga <Emphasis Type="Italic">Ecklonia stolonifera</Emphasis> in Genetically Diabetic KK-A<Superscript>y</Superscript> Mice

The dietary intake and control of blood glucose levels are very important in hyperglycemic patients and α-glucosidase inhibitors are a cost-effective means to preventing the progression of diabetes. In search of a natural inhibitor from food materials, α-glucosidase inhibitory activity and the anti-hyperglycemic effects of a brown alga, Ecklonia stolonifera, were investigated using non-insulin dependent diabetic mice. Methanolic extract of E. stolonifera (MEE), which contains a high content of polyphenols, showed strong inhibition of α-glucosidase in vitro. Male KK-A^y mice, a genetically non-insulin dependent diabetic model, showed hyperglycemia with aging, but the ingestion of MEE suppressed the increase in plasma glucose and lipid peroxidation levels in unfasted KK-A^y mice dose dependently. In KK-A^y mice, which were fed the MEE diet for 4 weeks, MEE moderated the elevation of plasma glucose levels after the oral administration of maltose. The polyphenols in MEE were estimated to be phlorotannins by HPLC-PDA and LC/MS analyses. These results demonstrate that E. stolonifera, seaweed typically used as a health food, has strong antidiabetic and antioxidant effects in vivo, thus, it may have beneficial properties in the prevention of diabetes and could be useful in the development of an antidiabetic pharmaceutical and functional food.     

Determination of Glycoalkaloids in Potatoes and Potato Products by Microwave Assisted Extraction

Potato glycoalkaloids can reach levels that are harmful to human health. A rapid and reliable microwave assisted extraction method for quantitative analysis of α-solanine and α-chaconine content in raw potato and potato based products is presented. A chemical microwave was used to determine optimal temperature and pressure conditions for the extraction of α-solanine and α-chaconine from Idaho grown tubers and six commercially available mashed potato products. Recovery efficiency of glycoalkaloids was 37% greater by microwave assisted extraction (19.92 mg/kg glycoalkaloid) as compared to conventional solid/liquid methods (12.51 mg/kg glycoalkaloid). Optimal extraction of glycoalkaloids from potato samples dissolved in methanol was achieved using a microwave reactor set to 90 °C for ten minutes. The interior of Idaho grown tubers was determined to contain lower levels of glycoalkaloids (19.92 mg/kg dry weight; 6.5 ± 1.78 mg α-solanine and 13.40 ± 1.65 mg α-chaconine), as compared to commercial potato products (33.86–81.59 mg/kg).