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.

     

Preparative isolation ofSolanum tuberosum L. glycoalkaloids by MPLC

Summary A new, efficient and economic method employing Medium Pressure Liquid Chromatography (MPLC) for the isolation of the two majorSolanum tuberosum L. glycoalkaloids (α-solanine and α-chaconine) is described. Potato peelings are homogenised with 5% acetic acid, the glycoalkaloids purified by filtration through an XAD-2 column and then by precipitation from the aqueous solution. The resulting glycoalkaloid fraction was purified by MPLC using a Silica Gel column and a CHCl₃:MeOH:2% NH₄OH mixture (70∶30∶5) as mobile phase to yield pure α-chaconine and a-solanine. This methodology can be used to obtain glycoalkaloids for enthomology and toxicological research where large amounts of these compounds are required.     

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.     

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.     

Unsaponifiable lipids from haulm and tuber sprouts of potato (Solanum tuberosum L.)

Summary Fractions containing Δ⁵- and Δ⁷-sterols, 4-methyl-sterols, triterpenic alcohols, tocopherols and hydrocarbons were isolated by TLC from petroleum ether extracts of the haulm and tuber sprouts of cv Désirée. Sterol and triterpenic alcohol fractions of unsaponifiable lipids of the haulm and tuber sprouts were found to contain twelve sterols and four triterpenic alcohols, respectively. 24R-4-stigmasten-3-on, Δ⁷, Δ⁷, lanosterol, cycloeucalenol and obtisufoliol have not been identified previously in unsaponifiable lipids from haulm and sprouts. In the hydrocarbon fractions of the haulm extract, C₂₃-to C₃₃-n-parafins, C₁₉-and C₃₁-cyclohexyl hydrocarbons, C₂₂-to C₃₈-olefins and squalene were identified, and in the hydrocarbon fractions of the tuber sprouts C₁₂-to C₂₆-olefins and squalene were identified.     

Isolation of solanidine from haulm and tuber sprouts of potato (Solanum tuberosum L.)

Summary The kinetics of enzymatic transformation of total glycoalkaloids (TGA) in haulm and tuber sprouts of potato to solanidine and the kinetics of solanidine isolation from plant material were studied. The optimum reaction conditions for the enzymatic TGA transformation were found for haulm to be: plant material to liquid phase 1:4 w/v, temperature 35°C, pH 5.4; and for tuber sprouts: plant material to liquid phase 1:3 w/v, temperature 37°C, pH 5.4. The. maximum degree of enzymatic transformation of TGA in haulm and tuber sprouts of 65 and 75%, respectively, was achieved after 50h. Carbon tetrachloride was the best solvent for solanidine extraction from the plant materials. The optimum ratio of plant material to carbon tetrachloride for the isolation of solanidine was 1:8 w/v for both extraction cycles, ensuring a yield of solanidine of approximately 55 and 65% from haulm and tuber sprouts, respectively, after 30 min.     

Amylase activity in potato tubers

Summary Amylase activity in extracts of sprouted tubers was optimised at final concentrations of soluble starch in the incubation medium of 0.6–2.0 mg cm⁻³. Optimum pH was 6. The exclusion of calcium ions from extraction and incubation media did not result in reduced enzyme activity. This, together with a shift in the absorption maximum of the starch-iodine complex almost identical to that observed with pure β-amylase, indicates the predominance of β-amylase in the extracts. Over a 15-min incubation period the linearity of the response was dependent upon the volume of tuber extract included in the assay medium. Gel filtration of extracts did not influence this response.     

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.     

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.     

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.     

Residues of free and bound maleic hydrazide in potato tubers

Summary The method of Vadukul (1991) for determining maleic hydrazide (MH) was modified and gave recoveries of free MH of 89%±4%. The values recorded on individual tubers ranged from 2–14 mg kg⁻¹. Maleic hydrazide was evenly distributed throughout the tuber (peel, outer and inner flesh) but concentration increased slightly as tuber size increased. The concentration of free MH decreased from 7 to 3 mg kg⁻¹ over the storage period of 5 1/2 months. Acid hydrolysis released substantial amounts of MH particularly from older potatoes (13 mg kg⁻¹) compared with 6 mg kg⁻¹ from new potatoes, implying that free MH is gradually converted to a bound form with time after treatment. No evidence was found for the presence of a β-glucoside of MH.     

Inhibitory Activity of Cinnamon Bark Species and their Combination Effect with Acarbose against Intestinal α-glucosidase and Pancreatic α-amylase

Inhibition of α-glucosidase and pancreatic α-amylase is one of the therapeutic approaches for delaying carbohydrate digestion, resulting in reduced postprandial glucose. The aim of this study was to evaluate the phytochemical analysis and the inhibitory effect of various cinnamon bark species against intestinal α-glucosidase and pancreatic α-amylase. The results showed that the content of total phenolic, flavonoid, and condensed tannin ranged from 0.17 to 0.21 g gallic acid equivalent/g extract, from 48.85 to 65.52 mg quercetin equivalent/g extract, and from 0.12 to 0.15 g catechin equivalent/g extract, respectively. The HPLC fingerprints of each cinnamon species were established. Among cinnamon species, Thai cinnamon extract was the most potent inhibitor against the intestinal maltase with the IC₅₀ values of 0.58 ± 0.01 mg/ml. The findings also showed that Ceylon cinnamon was the most effective intestinal sucrase and pancreatic α-amylase inhibitor with the IC₅₀ values of 0.42 ± 0.02 and 1.23 ± 0.02 mg/ml, respectively. In addition, cinnamon extracts produced additive inhibition against intestinal α-glucosidase and pancreatic α-amylase when combined with acarbose. These results suggest that cinnamon bark extracts may be potentially useful for the control of postprandial glucose in diabetic patients through inhibition of intestinal α-glucosidase and pancreatic α-amylase.     

Antioxidative and Anticancer Activity of Extracts of Cherry (<Emphasis Type="Italic">Prunus serrulata</Emphasis> var. <Emphasis Type="Italic">spontanea</Emphasis>) Blossoms

Organic solvent (methanol, ethanol, and acetone) extracts and water extracts of cherry (Prunus serrulata var. spontanea) blossoms were prepared, and antioxidant activities of the extracts were evaluated. Methanolic CBE (100 μg/ml) showed the highest total phenol content (104.30 μM), radical scavenging activity (34.2%), and reducing power (0.391). The effect of CBE on DNA damage induced by H₂O₂ in human leukocytes was evaluated by Comet assay. All CBE was a potent dose dependent inhibitor of DNA damage induced by 200 μM of H₂O₂, methanolic CBE showed the most strong inhibition activity. The methanolic CBE of 500 μg/ml showed 38.8% inhibition against growth of human colon cancer cell line HT-29. These results indicated that cherry blossoms could provide valuable bioactive materials.     

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.     

Antioxidant Activities of Four Edible Seaweeds from the Southern Coast of Thailand

The aqueous extracts of four marine algae, Caulerpa racemosa var. macrophysa, Gracilaria tenuistipitata var. tenuistipitata, Sargassum sp., and Ulva lactuca, from the coastal areas in Southern Thailand, were prepared by boiling dried seaweed powder in water for 3 h, and by autoclaving each sample at 120 °C for 3 h. They were then freeze-dried and evaluated for their antioxidant activities using DPPH (1, 1-diphenyl-2-picrylhydrazyl), hydroxyl radical (OH^•) and superoxide anion (O₂^•−) scavenging assays. Boiling extracts of the seaweeds, except C. racemosa, were found to have higher total phenolic contents (TPC) than those obtained from the autoclave method. The antioxidant results also showed that O₂^•− scavenging activity existed only in the boiling extracts of C. racemosa, G. tenuistipitata, and U. lactuca. In DPPH and OH^• assays, however, almost all the boiling extracts were less active than the autoclave ones. Among the four alga species, Sargassum sp. was the most active. Both extracts of this seaweed had the highest TPC and also displayed the strongest DPPH^• and OH^• inhibitory activities. A strong positive-correlation between the antioxidant potency and TPC of the autoclave extracts was found, while for the boiling extracts such relation was very weak. This result thus reflected that in addition to the phenolic compounds, there might be some other active components present in these extracts involved in the antioxidant activity.     

Antioxidant activity and phenolic content of chestnut (Castanea sativa) shell and eucalyptus (Eucalyptus globulus) bark extracts

Chestnut (Castanea sativa) shell and eucalyptus (Eucalyptus globulus) bark, waste products of the food and wood industries, respectively, were analysed as potential sources of antioxidant compounds. The extraction yield, the antioxidant activity and total phenols content of the extracts were greater in chestnut shell than in eucalyptus bark for most of the extraction conditions essayed. Extraction of chestnut shell with a 2.5% Na₂SO₃ aqueous solution led to the highest extraction yield, 25.6%, total phenols, 13.4 g gallic acid equivalent/100 g oven-dried shell, and FRAP antioxidant activity, 80.7 mmol ascorbic acid equivalent/100 g oven-dried shell. Extraction with methanol:water (50:50, v/v) provided the best results for eucalyptus bark. The antioxidant activity and the total phenols content of the extracts had a positive linear correlation. FTIR spectroscopy confirmed the higher content of phenolic compounds in chestnut shell extracts compared to eucalyptus bark extracts. Chestnut shell extracts were characterized by the presence of high molecular weight species whereas lower molecular weight species were predominant in eucalyptus bark extracts.     

Biochemical Composition and Indigestible Oligosaccharides in Phaseolus vulgaris L. Seeds

Common beans have a high nutritional value, but contain galactooligosaccharides (GO), which cause flatulence and intestinal discomfort in humans. The biochemical composition of ten bean cultivars was determined to select those of high protein and low GO contents. The cultivars varied in carbohydrate (47.02–60.17%), GO (3.12 – 5.71%), protein (22.17–33.50%), lipid (1.13–1.81%), moisture (11.42–12.93%) and ash contents (4.08–5.61%). ‘Mexico 222’ presented the highest α-galactosidase activity. Protein and GO contents were positively correlated. ‘Perry Marrow’ combined high protein and low GO concentrations, indicating it can be used in improvement programs aiming at high-quality cultivars for human consumption.     

Studies on Indian Green Leafy Vegetables for Their Antioxidant Activity

To identify the potential of green leafy vegetables (GLV) as antioxidants, methanolic extracts of Amaranthus sp., Centella asiatica, Murraya koenigii and Trigonella foenum graecum were studied for their antioxidant activity in different systems at multiple concentrations. Total antioxidant activity assessed by phosphomolybdenum method, free radical scavenging activity by 1,1-diphenly-2-picryl hydrazyl (DPPH), reducing power and ferrous ion chelating activity were determined. The GLV were analyzed for ascorbic acid, total and β-carotene and total polyphenol contents. The ascorbic acid, total carotene, β-carotene and total phenolic content (tannic acid equivalents) of the GLV ranged between 15.18–101.36, 34.78–64.51, 4.23–8.84 and 150.0–387.50 mg/100 g GLV, respectively. The extracts were found to have significantly different levels of antioxidant activities in the systems tested. The total antioxidant activity was highest in Murraya koenigii (2,691.78 μmol of ascorbic acid/g sample) and least in Centella asiatica (623.78 μmol of ascorbic acid/g sample). The extract concentration causing 50% inhibition of DPPH (IC₅₀) was determined (M. koenigii < C.asiatica < Amaranthus sp. < T. graecum). The maximum DPPH scavenging activity and reducing power was exhibited by Murraya koenigii. Multiple regression analysis showed that the relationship of total antioxidant activity, free radical scavenging activity, and reducing power with polyphenol and total and β-carotene was highly significant. Paper awarded the Young Scientist Award in Experimental Nutrition at the Young Scientist Award Session of the 39th Annual Meet of the Nutrition Society of India, Hyderabad, India on 15–16th Dec 2007.     

Die Effizienz der Nassfäuleabwehr bei der Wundheilung von Kartoffelknollen. 1. Bestimmung der befallsauslösenden Erregerdichte und ihrer Umweltbeeinflussung

Zusammenfassung Nach Inokulation mit abgestuften Bakteriendichten wurde über eine Vitalf?rbung von Knollengewebe der Schwellenwert zur F?uleausl?sung bestimmt. Seine Erh?hung nach 24 h Wundheilung charakterisiert die Effizienz der biochemischen F?uleabwehr. Die Abwehrreaktionen des Knollengewebes und der Prozess der F?uleausl?sung sind in unterschiedlichem Masse von der Temperatur, der Feuchtigkeit und dem O₂- sowie dem CO₂-Gehalt der Atmosph?re abh?ngig.

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Summary The inoculum densities (effective dose) ofErwinia carotovora sub sp.atroseptica (Eca) needed to initiate infection of tuber tissue before and after a 24 h wound-healing period, was determined using a vital stain. The effects of various treatments on the efficiency of the defence reaction associated with wound healing, and on rot initiation were manifested as changes in the effective dose. The defence reaction was inhibited by temperatures <10°C, reduced O₂ levels ≤5%, and CO₂ proportions ≥20% (Fig. 3a and 4a). Moisture loss in stored tubers increased their defence reaction compared to that of freshly harvested ones (Table 1). The effect of these factors on Eca development was identical to that which occurs in potato production where tuber wounds are generally contaminated with Eca. Rot initiation was best prevented by removal of water (Table 1), storage at 15°C, avoidance of reduced O₂ levels <10% and CO₂ accumulation (Fig. 3b and 4b).

     

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.