Seed Alkaloid Content Research Articles

Quantitative and qualitative content of alkaloids in seeds of a narrow-leafed lupin (Lupinus angustifolius L.) collection

Alkaloids represent the main antinutritional factor in lupins. The total content and qualitative composition of four major alkaloids in L. angustifolius L. were analyzed. The material included 329 accessions from the Polish collection divided into three classes of origin: wild lines, cultivars, and other man-made accessions. A very broad differentiation was found in terms of total alkaloid content—from 0.0005 to 2.8752 % of seed dry weight. In most cases, cultivars were characterized by a sharply decreased content of alkaloids, even below 0.01 % of seed dry weight. The average proportions of individual alkaloids were also very differentiated: lupanine—0.98 to 73.0 % of total alkaloid content (mean 46.4 %), 13-hydroxylupanine—15.6–71.1 % (mean 35.6 %), angustifoline—0–49.8 % (mean 15.5 %) and isolupanine—0–34.0 % (mean 2.5 %). The above mean values are probably typical for this species. In some accessions, lupanine and 13-hydroxylupanine accounted for 90–100 % of all major alkaloids. The average content of isolupanie (2.5 % of all alkaloids) allows it to be consider a major alkaloid of L. angustifolius, but quite frequently a ratio below 1 % or even its absence was stated. The three classes of origin were divided into three significantly different groups based on total alkaloid content as well as individual alkaloid content. Among wild lines, high alkaloid accessions were most numerous, but among cultivars it was low alkaloid accessions. The last class also contained numerous accessions with the lowest content of individual alkaloids. The influence of the content of individual alkaloids on total alkaloid content was also investigated in the wild lines and cultivars.

Studies on the alkaloid composition of the Hawaiian Baby Woodrose Argyreia nervosa, a common legal high

Seeds from the Hawaiian Baby Woodrose Argyreia nervosa of different origin and labelling and with allegedly high levels of ergot alkaloids were analysed using high performance liquid chromatography–high resolution mass spectrometry (HPLC–HRMS/MS) technique. Lysergic acid amide (LSA), ergometrine, lysergol/elymoclavine/setoclavine, chanoclavine, lysergic acid and their respective stereoisomers were identified as well as penniclavine and lysergic acid α-hydroxyethylamide. In addition, methylergometrine, methysergide, and lysergylalanine were detected, some high molecular weight ergot alkaloid derivatives and hydroxyalanine derived ergopeptide fragments were detected indicating the presence of ergopeptides in the seeds. The results of the study demonstrate that the content of ergot alkaloids in Argyreia nervosa seeds depends on the quality of the material. For a consumer the quality of the seeds is unforeseeable. For the toxicological expert it is essential to investigate not only the identity of such a confiscated seed material, but also the various ergot alkaloid constituents to assess the hazardous nature and the toxic potential of the material.

Alkaloid and flavonoid rich fractions of fenugreek seeds (Trigonella foenum-graecum L.) with antinociceptive and anti-inflammatory effects

The seeds of fenugreek (Trigonella foenum-graecum L.) have medicinal uses as hypoglycemic, antinociceptive and anti-inflammatory agents. We aimed to evaluate the antinociceptive and anti-inflammatory effects of the major fractions of fenugreek seeds. The methanolic extract of the plant seeds was partitioned using a liquid–liquid extraction procedure to give six major fractions. Following phytochemical screening of isolated fractions, the total extract and each fraction were evaluated for their antinociception and anti-inflammatory effects using formalin and carrageenan-induced paw edema tests respectively. The methanolic extract exhibited both antinociceptive and anti-inflammatory effects at a dose of 100mg/kg. Among the tested fractions, alkaline chloroform fraction (AKC), which was alkaloid positive in screening tests, showed the most anti-nociceptive effect in a dose-dependent manner. AKC fraction was as effective as morphine (5mg/kg) in this regard. Both aqueous and acidified chloroform fractions (ACC) could significantly inhibit paw edema at a different dose. The latter fraction dose-dependently inhibited carrageenan-induced paw edema. The results of phytochemical screening tests confirmed the presence of flavonoids in both ACC and aqueous fractions. It can be concluded that the alkaloid and flavonoid content of fenugreek seeds can be responsible for antinociception and anti-inflammatory effects of the plant respectively.

Simultaneous determination of atropine and scopolamine in different parts of Hyoscyamus arachnoideus Pojark plants by high-performance liquid chromatography (HPLC)

A reverse phase high-performance liquid chromatography (HPLC) equipped with UV-PDA detector was used for the analysis of main tropane alkaloids, scopolamine and atropine, in the roots, aerial parts and seeds of Hyoscyamus arachnoideus Pojark plants collected from five different parts of Iran. Results showed that scopolamine was only the main alkaloid of two H. arachnoideus populations seeds, while atropine was the major alkaloid of almost all parts, especially roots. The highest and lowest content of tropane alkaloids was observed in populations from Zangane village and Ashtian Mountain, respectively. The alkaloid content of seeds was in general more than other parts tested. The linearity of the method was in the range of 4 to 400 µg/ml for atropine and 0.8 to 80 µg/ml for scopolamine. Limit of detection (LOD) and limit of quantification (LOQ) values were 5.15 and 17.4 µg/ml for atropine and 1.92 and 6.4 µg/ml for scopolamine.

THE EFFECTS OF DIFFERENT DEBITTERING METHODS ON THE PRODUCTION OF LUPIN BEAN SNACK FROM BITTER LUPINUS ALBUS L. SEEDS

ABSTRACT The aim of this study was to remove the alkaloids from bitter lupin (Lupinus albus L.) seeds by different debittering methods and to produce a lupin bean snack. During production of the snack, the total alkaloid content of seeds decreased significantly (P ≤ 0.01) from 14.4 g/kg to a level that was undetectable. A major alkaloid in lupin seeds was identified as lupanine. It was present at a concentration of 12.513 g/kg, which constituted 87% of the total alkaloid content. The other alkaloids that were identified were sparteine, albine, α‐isolupanine and multiflorine. The concentrations of the alkaloids decreased gradually during the debittering process. The values for dry matter, protein, lipid, starch and ash content of seeds changed in a statistically significant manner (P ≤ 0.05, P ≤ 0.01) during production of the snack. Aqueous debittering at room temperature was found to be a more favorable process because it resulted in more acceptable sensorial properties than alkaline and thermal debittering methods.PRACTICAL APPLICATIONSBitter lupin seeds (Lupinus albus L.) cannot be consumed directly because they naturally contain high amounts of toxic alkaloids, which cause bitterness, despite their high protein content. The aim of this study is to remove bitterness from lupin seeds with different debittering methods and to produce a lupin bean snack.

Effects of Temperature on the Alkaloid Content of Seeds of Lupinus angustifolius Cultivars

AbstractThree experiments were conducted to investigate the effects of high temperatures during seed filling on the alkaloid content of narrow‐leaf lupin cultivars. Six cultivars of Lupinus angustifolius were grown in field experiments under different weather conditions in four subsequent years. A high content of alkaloids was found in the seeds harvested in 2006, in which the growing season was characterized by high ambient temperatures during seed filling. A second experiment was performed in the green house at different temperatures (10, 20 and 30 °C) using one cultivar in 2006. This experiment confirmed the results of the field experiments as higher temperatures resulted in a higher alkaloid content of the seeds. In a third temperature stress experiment, three cultivars were grown under long day conditions at day/night temperatures of 30 °C/16 °C and 20 °C/16 °C in growth chambers in comparison with an outdoor control at mean temperatures of 15.5 °C. Like in the other experiments, the seed alkaloid content increased with rising temperature. From these results, it may be concluded that the seed alkaloid content is strongly influenced by the temperature during initiation of flowering up to pod ripening. This has to be taken into account e.g. in trials for cultivar release in which the alkaloid content is a knock‐out criterion.

A quinolizidine alkaloid O‐tigloyltransferase gene in wild and domesticated white lupin (Lupinus albus)

AbstractWild white lupins have high levels of alkaloids, which cause a bitter taste, whereas domesticated white lupin varieties have a very low content of alkaloids in seeds. Genes for bitterness from wild white lupins are a contamination threat to domesticated white lupin via cross‐pollination. The gene(s) for alkaloid synthesis have not been clearly identified, and the associated molecular background among wild white lupin, domesticated and contaminated domesticated plant materials is unknown. So far, only tigloyl‐CoA:(−)‐13alpha‐hydroxymultiflorine/(+)‐13alpha‐hydroxylupanine O‐tigloyltransferase (HMT/HLTase) cDNA has been cloned based on protein analysis, which was suggested as encoding a quinolizidine alkaloid transferase regulating quinolizidine alkaloid biosynthesis. This gene has not yet been well characterised in important white lupin genotypes. In this study, we found that the majority of the intron sequence of the HMT/HLTase gene differed between wild white lupin accessions P25758 and P27593, and between the commercial varieties. The expression pattern as well as the expression level of the HMT/HLTase gene showed no difference between the P25758 and the low‐alkaloid variety Kiev mutant, suggesting the expression of the HMT/HLTase gene has no correlation with bitterness. However, the intron sequence is useful as a DNA marker in the identification of the contamination source of bitter seeds in commercial lupin seed lots.

Treatment of Lupin Seed Using the Instantaneous Controlled Pressure Drop Technology to Reduce Alkaloid Content

Despite their high protein content, lupin seeds are not as fully utilised as some other grain legumes. The drawback in their utilisation is mainly due to the presence of water soluble, poisonous alkaloids. In this study, an instantaneous controlled pressure drop (DIC) treatment followed by an aqueous extraction was performed on lupin seeds. Two species Lupinus albus and Lupinus mutabilis with different initial total alkaloid content of 0.025% and 5.5% (d.b.), respectively have been studied. Lupanine was the major alkaloid found in both lupin species. Experimental results proved the feasibility of using DIC treatment to reduce the alkaloid content of lupin seeds. Optimised DIC treatment combined with an adequate 2h soaking in water reduced the alkaloid content from 5.5% to 2.2% of L. mutabilis and from 0.025% to 0.0075% of L. albus.

Influence of chemical characteristics of soil on mineral and alkaloid seed contents of Colchicum autumnale

Alkaloids from meadow saffron seeds are used in numerous medicines. Variations of seed alkaloid content (colchicine and colchicoside) were observed between natural sites. Nevertheless, this character remained stable when these accessions (corms collected on different natural sites) were cultivated in a same field. A relation between soil properties and seeds alkaloid content can be expected. Soil analyses were performed in three soil layers (0–30, 30–60, 60–90 cm) on three meadow saffron natural sites and two cultivated sites. Linear regressions were performed between 20 soil factors, analysed in the three soil layers and the seed alkaloid content. Cobalt and calcium soil contents were positively correlated with seed alkaloid content. The coefficients of determination ( R 2) were the highest in the 30–60 cm soil layer, respectively, of 0.85 and 0.74. Mineral analyses were performed on seed. In spite of the important difference of level, linear relations were observed between the sum of Ca and 1000 × Co contents (Ca + Co) in seeds and in soil ( R 2 = 0.76). A highly significant relation was shown between (Ca + Co) in seeds and alkaloid content ( R 2 = 0.96) showing for the first time on Colchicum autumnale the positive influence of these mineral elements on the alkaloid synthesis. Soil and foliar supplies of Ca and Co could be tested to improve seed alkaloid content.

Seed yield and components of alkaloid of meadow saffron (Colchicum autumnale) in natural grassland and under cultivation

Alkaloids extracted from meadow saffron seeds are used in numerous medicines. To test the influence of cultivation and climatic conditions on the production and the quality of meadow saffron seeds, we studied seed alkaloid yield on a natural site (grassland) and in a cultivated crop over several years. Seeds contained in capsules harvested before their dehiscence were dried and ground. The alkaloids of the extracts (colchicoside, demethylcolchicine and colchicine) were analyzed by high-pressure liquid chromatography. Seed alkaloid content was not influenced by year, inter-plant competition, or number of capsules per plant. Seed dry matter yield per plant, however, practically tripled in the cultivated crop due to an increase in the number of capsules per plant compared to the grassland. In the cultivated crop, 47% of fruiting plants produced two new plants (corms) every year. Four years after planting, this vegetative multiplication resulted in intraspecific competition, inducing a decline in seed yield per plant linked to a decrease in the weight of seeds per capsule. We were able to show that cultivated production of meadow saffron greatly increased dry matter and alkaloid yield per plant and per hectare, but that it could lead to the need to thin after several years. Key words: meadow saffron, alkaloids, colchicine, cultivation

Effect of magnesium and nitrogenous fertilisers on the growth and alkaloid content in Lupinus angustifolius L.

The effect of kieserite (MgSO4.H2O) and nitrogenous fertilisers in various forms [control, Ca(NO3)2, (NH4)2SO4, NH4NO3, CO(NH2)2] on the growth and alkaloid content in L. angustifolius was investigated. Mineral fertilisers were applied at doses of 60 kg/ha of Mg and 75 kg/ha of N before sowing. Kieserite did not have a significant influence on the growth and dry matter production of lupin. Ca(NO3)2 and NH4NO3, although increasing the yield of straw, did not affect the yield of seeds. The application of (NH4)2SO4 decreased the yield of straw and seeds when compared with the effect of the above-mentioned fertilisers. The effect of Mg and nitrogenous fertilisers on alkaloid biosynthesis in vegetative organs depended, most of all, on the weather conditions in the vegetation period and on the developmental stage of lupin at which they were applied. At the stage of 6-7 leaves, the alkaloid content was negatively correlated with the content of Mg in plants (R2 = 67%). The influence of mineral forms of nitrogen on the alkaloid content in the dry matter increased in the following order: (NH4)2SO4 <�NH4NO3 <�Ca(NO3)2. The nitrogen fertilisers studied significantly modified the alkaloid content in the seeds, but the effect of nitrogen fertilisers also depended on Mg. In the treatments without Mg, nitrogenous fertilisers significantly increased the content of alkaloids by 9-17%. The greatest increase in alkaloid content was found when NH4NO3 was applied, followed by (NH4)2SO4. Simultaneous application of Mg and nitrogen decreased the content of alkaloids in seeds, depending on the year of study, by 2-8%.

Electronically monitored cowpea aphid feeding behavior on resistant and susceptible lupins

AbstractThe feeding behavior of cowpea aphid, Aphis craccivora Koch (Homoptera: Aphididae) was examined on seedlings of narrow leafed lupin, Lupinus angustifolius L., and yellow lupin, L. luteus L., using electronic monitoring of insect feeding behavior (EMIF). Aphid feeding behavior was first compared between resistant (cv. Kalya) and susceptible (cv. Tallerack) varieties of narrow‐leafed lupin. Aphids spent significantly more time in non‐ penetration and stylet pathway activities, and significantly less time in the sieve element phase on Kalya than on Tallerack, suggesting that feeding deterrence is an important component of aphid resistance in Kalya. Aphid feeding on a susceptible yellow lupin variety (cv. Wodjil) was then compared with that on two resistant lines, one (Teo) with high and the other (94D024‐1) with low seed alkaloid content. There were no consistent differences in aphid feeding behavior between Wodjil and Teo. Total, mean and percentage sieve element phase times were significantly lower, and total and percentage times in non‐phloem phase were greater on 94D024‐1 than on Wodjil, suggesting the possibility of phloem‐based deterrence in 94D024‐1.

Size and Alkaloid Content of Seeds in Induced Autotetraploids of Datura innoxia, Datura stramonium and Hyoscyamus niger

Alkaloid content and metric features of seeds in diploid and induced autotetraploid Datura innoxia Mill., D. stramonium L. and Hyoscyamus niger L. were studied. As a result of chromosome doubling, the seeds of tetraploid forms were larger and heavier. Scopolamine was the main alkaloid in both ploidy levels of D. innoxia and H. niger whereas hyoscyamine was the main alkaloid in D. stramonium. The tetraploid seeds of Datura innoxia, D. stramonium and Hyoscyamus niger had 1.86-, 1.65- and 1.96-times higher alkaloid contents compared to the seeds of the diploid form. The scopolamine to hyoscyamine ratio in tetraploids was higher in both Datura species and lower in Hyoscyamus niger. The increased size and alkaloid content makes tetraploid seeds a better source of alkaloids than diploid seeds.

Effect of drought stress on content and composition of seed alkaloids in narrow-leafed lupin, Lupinus angustifolius L.

The effect of drought stress at three growth stages on seed alkaloid content in two sweet (W26 and Polonez) and one bitter variety (Zubr) of spring sown Lupinus angustifolius L. was investigated in a lysimeter experiment on sandy soil in Denmark. Full irrigation resulted in medium alkaloid content. Drought stress during the vegetative phase increased alkaloid content to 0.094, 0.192 and 2.55% of seed dry matter, respectively. At the flowering stage alkaloid content decreased to 0.070, 0.152 and 1.99%, while drought stress during pod-filling increased alkaloid content for the determinate variety W26 to 0.097%, but decreased it to 0.161 and 1.98% for the two indeterminate varieties Polonez and Zubr, respectively. Drought during either flowering or pod-filling resulted in an increased mean seed weight and a reduced number of seed per square metre. Alkaloid content was negatively correlated to seed weight. The alkaloid composition differed for the three genotypes, and was affected by drought treatment. Compared to the bitter variety the sweet varieties were characterized by reduced concentrations of lupanin, 13-hydroxylupanin and angustifolin, whereas spartein, isolupanin and isoangustifolin were little affected.

Purine alkaloids in seeds ofTheobroma species from the Amazon

The content of purine alkaloids in seeds of fourTheobroma species from the Amazon basin was analized by a reversed phase high-performance liquid chromatographic method.Theobroma grandiflorum was found to contain 0.38% of 1,3,7,9-tetramethyluric acidT. subincanum 0.07% 1,3,7,9-tetramethyluric acid,T. speciosum 0.01% theobromine and 0.04% 1,3,7,9-tetramethyluric acid and finallyT. mariae 2.35% theobromine and 0.12% caffeine. Theophylline could not be detected in any of these species. The results were confirmed qualitatively by thin-layer chromatography.

Alkaloids and phenolics of Colchicum turcicum

The alkaloid content (3-demethylcolchicine, colchifoline, N-deacetyl- N-formylcolchicine, colchicine, cornigerine, 2-demethyldemecolcine, 3-demethyldemecolcine, demecolcine) of corms, leaves and seeds of C. turcicum was determined by HPLC. In addition to these compounds, corms contained β-lumicolchicine, luteolin, and vanillic acid. Complete spectral data of colchifoline are presented.