Alliin Content Research Articles

Influence of Fertilizer Practices on S-Containing Metabolites in Garlic (Allium sativum L.) under Field Conditions

Cysteine sulfoxides (e.g., alliin) are the characteristic sulfur-containing secondary compounds in garlic, which account for taste and pharmaceutical quality. It was the aim of the present study to investigate the influence of sulfur and nitrogen supply under field conditions on the alliin content and cysteine and glutathione as possible precursors. Sulfur and nitrogen were applied in four different rates, and five samplings were conducted. Sulfur fertilization significantly increased the cysteine, glutathione, and alliin contents of leaves and bulbs, while nitrogen fertilization had no significant influence. Cysteine increased by a factor of 1.3-1.5 in leaves and 1.0-2.0 in bulbs. Glutathione increased significantly in bulbs by a factor of 0.9-1.6 but only at main growth and not at maturity. The alliin concentration in bulbs increased with S fertilization significantly at all harvesting dates and at maturity from 5.1 to 11.2 mg g(-1) of dry weight. High sulfur application in combination with low nitrogen fertilization increased the alliin concentration in garlic significantly during main growth until the beginning of ripening. At the last harvest, 15 kg ha(-1) S resulted in high-quality garlic suitable for consumption and use in plant protection or pharmaceutical industries.

Total sulphur and organosulphur compounds in garlic and ramsons plant organs at the end of vegetative period

In this study was analysed total sulphur and content of organosulphur compounds such alliin, diallyl disulfide (DD), reduced glutathione (GSH) and L-cysteine in the bulb and leaves of garlic and ramsons at the end of vegetative period. Total sulphur content was determined by ion chromatography in the form of sulphate ion. Analysis of alliin, DD, GSH and L-cysteine was performed by HPLC using UV-VIS, electrochemical and fluorescence detectors. Sulphur content (mg/g) in leaves and bulb of garlic was: bulb 0,63 and leaves 0,66. Content of alliin: bulb 4,8×10–2µg/g, leaves 3,8×10–2µg/g, and DD: 12,97mg/g, but in the leaves of garlic DD was below of the limit of detection. Content of L-cysteine in the bulb of garlic was 15,82mg/g, leaves 2,31mg/g, while the content of GSH in the bulb of garlic was 21,9mg/g and leaves 12,69mg/g. Total sulphur content (mg/g) in leaves and bulbs of ramsons at the end of vegetative period was: bulb 0,93 and leaves 0,74. Content of alliin: bulb 23,2×10–2µg/g, leaves 7,3×10–3µg/g. The content of DD in the bulb of ramsons was 1,78mg/g, while in the leaves content of DD was below of limit of detection. L-cysteine in the bulb of ramsons was 14,51mg/g and leaves 0,94mg/g, GSH in the bulb of ramsons was 14,51mg/g and leaves 8,94mg/g. In general the contents of total sulphur and organosulphur compounds in the bulb of garlic and ramsons are higher than in leaves.

Alliin production in various tissues and organs of Allium sativum grown under normal and sulphur-supplemented in vitro conditions

Garlic extract contains alliin (S-allyl-l-cysteine sulfoxide), an organosulphur compound that contributes to its therapeutic value and pharmacological importance. Alliin production in plant organs (leaf and root), plantlet, callus (non-embryogenic and embryogenic), embryo (proliferated, matured and germinated embryos), grown under in-vitro conditions was evaluated after 8 weeks. Highest alliin content was recorded in leaves. Clove callus and germinated embryos also showed alliin production. Evaluation of alliin content of in-vitro grown tissues both in normal (control) and sulphur supplemented conditions (4, 8, 16, 32 mg l−1) showed that sulphur treatment at supply of 16 mg l−1 gypsum (CaSO4) significantly enhanced the production of alliin content in all in-vitro grown tissues and organs. These findings suggest that production of alliin is dependent upon type and age of tissue and can be altered by growth conditions.

Improved alliin yield in somatic embryos ofAllium sativumL. (cv. Yamuna safed) as analyzed by HPTLC

Direct somatic embryo formation (without intervening callus) from garlic clove basal tissue was induced in which the influence of plant growth regulators (PGRs) on various explants was examined. Medium added with 2.0 mg/l 6-benzylaminopurine (BAP) and 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) were the most effective PGR combination for somatic embryo induction. It induced embryos directly in 85.5% of the basal clove explant. Callus induction was also obtained from other parts of explant and 2.0 mg/l 2,4-D induced callusing in 86.5% of the inoculated explants. Protein, amino acid and alliin content were measured in callus and in embryos. Somatic embryos had more soluble protein and free amino acid compared to callus. HPTLC analysis revealed that alliin was significantly high in somatic embryos compared to undifferentiated callus tissue; the content was even more in older embryos. The present study of Allium indicates that the event of morphogenetic development including in vitro embryogeny can effectively be analysed by monitoring the changes of biochemical profiles.

Immobilization of alliinase and its application: Flow injection enzymatic analysis for alliin

Alliinase was immobilized on chitosan microspheres by means of glutaraldehyde, and a flow-injection enzymatic analytical system was developed for determination of alliin based on the immobilized alliinase and an ammonia gas electrode. The factors affecting the activity of the immobilized enzyme, such as glutaraldehyde concentration, cross-linking time and the amount of alliinase, were investigated. Results showed that the maximum of enzyme activity could be obtained at glutaraldehyde concentration of 4%, cross-linking time of 2 h and the amount of alliinase 20.2 u. The properties of the immobilized alliinase were also studied in detail. For the immobilized alliinase, the highest activity was allowed at pH of 7.0 and temperature at 35°C. Besides, the immobilized enzyme showed good thermal and pH stabilities. The flow-injection enzymatic analytical system based on the immobilized alliinase and an ammonia gas electrode provided linearity in the 1 × 10-5 to 1 × 10-3 mol/L alliin concentration range and exhibited good repeatability and operational stability.

Effect of consuming raw or boiled garlic on agonist‐induced ex vivo platelet aggregation: an open trial with healthy volunteers

Prior clinical trials with garlic powder (0.6–0.9 g dry wt/d; alliin = 8–12 mg) as tablets have shown inconsistent effects on induced platelet aggregation, using platelet rich plasma (PRP). No trials have been reported for raw or cooked garlic. The objective of this open trial was to determine if a high dose (near the maximum tolerable) of raw garlic (8 g fresh wt/d = 2.8 g dry wt; allicin content = 35 mg, from 88 mg alliin) or boiled garlic (8 g fresh wt/d; no allicin, alliin content = 84 mg), consumed in a sandwich, are able to inhibit induced platelet aggregation in 4 weeks in 9 healthy volunteers who showed normal antiaggregatory effects with aspirin (325 mg). Optical density was used to measure aggregation in PRP; electrical impedance was used for whole blood (WB). Low agonist concentrations were used to maximize antiaggregatory effects: collagen at 0.2 & 0.5 μg/mL (PRP) and 0.5 & 1.0 μg/mL (WB), ADP at 1.8 & 2.5 μM (PRP) and 5 & 12.5 μM (WB), and epinephrine at 1 μM (PRP). Results: raw garlic significantly decreased PRP aggregation to a small extent (5–8%) with all agonists, but it significantly increased collagen-induced WB aggregation (10–13%). Boiled garlic had no effects. Conclusions: the results are method dependent, but considering the large dose used, it appears unlikely that consumption of normal levels of raw or boiled garlic (1–3 g) will have an effect on ex vivo aggregation in healthy individuals. Funded by NIH/NCCAM.

Genotype, nitrogen fertility and sulphur availability interact to affect flavour in garlic (Allium sativum L.)

SummaryExperiments were carried out to investigate the influence of combined nitrogen (N) and sulphur (S) fertility on the organosulphur compound content of garlic bulbs, especially alliin, as a specific flavour quality trait related to the health-value of garlic. Three cultivars, ‘Printanor’, ‘Morasol’ and ‘Messidrôme’, were grown in the greenhouse and in vitro. Sulphur was increased in the macronutrients within the range 0 – 144 mg l–1. Numerous combinations of both S and N were tested in vitro. In the greenhouse, increasing N levels, from 184.8 mg l–1 to 856.8 mg l–1, were tested with a single high level of S (128 mg l–1). Increasing S alone did not affect the growth or bulb weight of any garlic cultivar, but slightly increased the alliin content of bulbs. No symptoms of S deficiency were observed under greenhouse conditions, although the same garlic cultivars grown in vitro at 0 mg S l–1 clearly suffered, their size being greatly reduced. This suggests an important role for S contamination, such as atmospheric S, in the process of S absorption by garlic plants, which is discussed. Increasing N levels significantly accelerated garlic maturity and decreased bulb yield. The effect of N on the organosulphur compound content of bulbs was cultivar-dependent, but alliin accumulation was not enhanced. ‘Printanor’ and ‘Morasol’ had the highest alliin levels and were negatively influenced by increasing N, while a slight synergistic effect of S and N was observed on ‘Messidrôme’. ‘Messidrôme’ also had the highest flavour potential when including precursors, and was more efficient in accumulating S into organosulphur compounds. It is therefore advisable to choose the most appropriate variety, and to adapt the N S fertilisation regime according to the environment, in order to produce garlic with the highest health-value.

Antiyeast Potency of Heated Garlic in Relation to the Content of Allyl Alcohol Thermally Generated from Alliin

ABSTRACT: The appearance of antiyeast activity in heated garlic extract was influenced by the temperature, heating time, and pH. The maximum antiyeast activity was attained at the temperature range between 110 and 130 °C for the heating time range of 90 and 30 min, respectively. Reduced antiyeast activity occurred above 130 °C. More potent antiyeast activity was generated when the pH of the garlic extract was adjusted to and below 4.0. The sensitivity to the antiyeast activity of heated garlic or heated alliin was quite different depending on the yeast strain. Species belonging to Zygosaccharomyces were the least sensitive, while Candida utilis and Pichia membranaefaciens were among the most sensitive. The alliin content of the garlic was in the range of 0.5% to 1.32%. When garlic extracts were heated at 121 °C for 120 min, the content of allyl alcohol generated from alliin through thermal degradation was in the range of 0.08% to 0.16%. The allyl alcohol produced upon heating was positively proportional to the content of alliin in garlic. The antiyeast potency expressed as the minimum inhibitory concentration (MIC) was negatively proportional to the content of allyl alcohol and thus to the content of alliin.

Inhibition of Carcinogen-Induced DNA Damage in Rat Liver and Colon by Garlic Powders With Varying Alliin Content

: The present study was designed to investigate the protective efficiency of three garlic powders, obtained from bulbs grown in soils with different levels of sulfur fertilization, against DNA damage. Increasing fertilization of soil resulted in an increased alliin content of the powders. Garlic powders were administered to rats for 2 weeks (5% of the diet) and their antigenotoxic effects were examined in the liver and the colon using the comet assay. Consumption of the different garlic powders induced a 35–60% reduction in DNA damage induced by N-nitrosodimethylamine (NDMA) in rat liver. Increased alliin content of the garlic powder was associated strongly with a proportional decrease in NDMA induced DNA alteration. DNA damage induced by aflatoxin B1 in the liver or by 1,2-dimethylhydrazine in the colon were also decreased strongly by the three garlic powders but these decreases were not correlated to the alliin content of the garlic powders. Feeding garlic powders did not modify the genotoxic activity of the direct-acting carcinogen methylnitrosourea in the colon. Part of our results supports evidence that fertilization can have an impact on the protective capacity of garlic bulbs.

Development and Validation of a Thin-Layer Chromatography-Densitometric Method for the Quantitation of Alliin from Garlic (Allium sativum) and Its Formulations

A selective, precise, and accurate high-performance thin-layer chromatographic (HPTLC) method has been proposed for the analysis of garlic and its formulations for their alliin content. The method involves densitometric evaluation of alliin after resolving it by HPTLC on silica gel plates with n-butanol-acetic acid-water (6 + 2 + 2, v/v) as the mobile phase. For densitometric evaluation, peak areas were recorded at 540 nm after derivatizing the resolved bands with ninhydrin reagent. The relation between the concentration of alliin and corresponding peak areas was found to be linear within the range of 250 to 1500 ng/spot. The method was validated for precision (interday and intraday), repeatability, and accuracy. Mean recovery was 98.36%. The method was applied for the quantitation of alliin in bulbs of Allium sativum Linn. (garlic) and its formulations. The proposed TLC method was found to be precise, specific, sensitive, and accurate and can be used for routine quality control of garlic and its formulations.

INFLUENCE OF ENVIRONMENTAL AND GENETIC FACTORS ON THE ALLIIN CONTENT OF GARLIC BULBS

The influence of environmental conditions and genotype x environment effects on the alliin content of garlic bulbs were studied. The objective was to improve the quality of garlic bulbs with regard to alliin content. Field trials were carried out for 2 years in France and Spain with increasing sulphur fertilisation in soil: 0-50-100-200 or 400 kg SO4/ha were investigated on 3 varieties in 2000 ('Printanor', 'Messidrome' and 'Morasol') and only on 'Printanor' in 2001. S fertilisation affected alliin content on 'Printanor' in France, but was not confirmed in Spain where alliin contents were 3 to 5 fold higher than in France. These results were correlated to the soil sulphur in the two areas as Spanish soil was always higher in water soluble and total sulphur than French soil. However, the influence of other factors could not be excluded. Experiments were also carried out in controlled conditions (in vitro and greenhouse) to study the influence of sulphur fertilisation combined to other environmental factors such as light, carbohydrate availability and temperature. A strong genotypic effect was confirmed in both controlled conditions with light spectral quality having a large effect on alliin content on the in vitro bulbs. Temperature did not have a large effect on allying, but did interact with sucrose concentration to affect alliin accumulation in the bulbs. We also found an interaction between S fertilisation and genotype in greenhouse, and genotype by light spectrum interaction, in vitro. Our results confirm that multiple factors affect alliin accumulation in garlic, and that these factors should be considered when growing garlic for flavour or therapeutic value. (Resume d'auteur)

(9) Compositional Characterization of a Garlic Germplasm Collection

A USDA germplasm collection of garlic (Alliumsativum) and related species (A. ophioscorodon, A. longicuspis) has been evaluated for horticultural characteristics and composition under California production conditions for several years. In 2004, 198 of the 217 accessions in the collection were evaluated. Bulbs were manually harvested late June to early July, cured about 3 weeks shaded at ambient temperatures, and the outer whorl of cloves manually peeled. Bulb and clove weights and percentage of dry matter were determined. Freeze-dried garlic powder was analyzed for alliin (precursor of flavor and health compounds) concentrations by HPLC. Thiosulfinates (mostly allicin, responsible for flavor and pungency) were determined by a spectrophotometric assay. Among the accessions evaluated in 2004, the percentage of dry matter ranged from 32.8% to 44.9%. Alliin content varied from 8.9 to 29.7 mg·g-1, and thiosulfinate concentrations ranged from 32.7 to 114.0 μmol·g-1. For comparison, the widely grown varieties California Early and California Late averaged 39.5% and 40.8% dry matter, 18.6 and 20.6 mg·g-1 alliin, and 68.1 and 78.8 μm·g-1 thiosulfinates.

Influence of Nitrogen and Sulfur Fertilization on the Alliin Content of Onions and Garlic

Onion (Allium cepa L.) and garlic (Allium sativum L.) were among the earliest cultivated crops and have been popular in folk medicine for centuries. Alliins (cysteine sulfoxides) are the characteristic sulfur (S) containing secondary metabolites of Allium species like onions, shallot, garlic, leek and chives and they cause taste and sharpness and are criteria for the pharmaceutical quality. The influence of the S nutritional status on the content of secondary S containing metabolites was shown for different crops such as oilseed rape, mustard, nasturtium, and allium species. It was the aim of this study to investigate the influence of the S and nitrogen (N) supply on the alliin content of onion and garlic and to evaluate the significance for crop quality. In a greenhouse experiment three levels of N and S were applied in factorial combinations of 0, 50, and 250 mg pot−1 S and 250, 500, and 1000 mg pot−1 N. 8 plants were grown in a Mitscherlich pot containing 8 kg sand. Leaves and bulbs were sampled twice during the growth period in order to follow up translocation processes. The first sampling was carried out when leaves were developed, but bulb growth had not yet started and the second one during main bulb growth. An increasing S supply was related to an increasing alliin content in leaves and bulbs of both crops, whereas nitrogen fertilization had only a minor influence. The alliin content in bulbs could be doubled by S fertilization. A translocation of alliin from leaves to bulbs was found so that time of harvest has a strong influence on the alliin content. At the beginning of plant development high alliin contents were found in leaves, while with bulb development they were translocated into this plant organ. The results show that the potential health benefits of Allium species could be distinctly improved by S fertilization.

Influence of Nitrogen and Sulfur Fertilization on the Alliin Content of Onions and Garlic

Influence of Nitrogen and Sulfur Fertilization on the Alliin Content of Onions and Garlic

Effects of garlic powders with varying alliin contents on hepatic drug metabolizing enzymes in rats.

The anticarcinogenic effect of garlic has been demonstrated in both epidemiologic and experimental studies. In this study, possible mechanisms involved in the anticarcinogenic effect of garlic consumption were assessed by determining its capacity to alter drug metabolizing enzymes, in relation with its alliin content. Rats were fed a diet for 2 weeks containing 5% garlic powders produced from bulbs grown on soils with different levels of sulfate fertilization and therefore containing differing amounts of alliin. Activities of several hepatic enzymes, which are important in carcinogen metabolism such cytochromes P450 (CYP) and phase II enzymes, were determined. Garlic consumption slightly increased ethoxyresorufin O-deethylase and CYP 1A2 levels. In contrast, garlic consumption decreased CYP 2E1 activity and the level of the corresponding isoform. UDP glucuronosyl transferase and glutathion S-transferase activities were increased by garlic powders. The alliin content of the garlic powders was positively correlated with UGT activity although not with other activities. Effects produced by garlic consumption were qualitatively similar to that of diallyl disulfide, a sulfur compound that has been extensively studied. These data could partially explain the chemoprotective effect of garlic.

Biosensoric detection of the cysteine sulphoxide alliin

Garlic (Allium sativum L.) and related species of the Alliaceae family are known for their cancer-protecting and antiatherosclerotic potential. The bioactivity of these plants is related to organo-sulphur compounds, mainly alliin. In the present investigation, an alliin-specific biosensor exploiting immobilised alliinase (EC 4.4.1.4) has been developed. Enzymatically formed ammonia was detected by a potentiometric sensor based on an ammonia electrode or a pH-sensitive electrolyte/insulator/semiconductor (EIS) layer structure made of Al/p-Si/SiO2/Si3N4. It could be demonstrated with both methods that this biosensoric approach yielded results comparable to sensitivities obtained by HPLC. Alliin concentrations down to 6×10−6M could be quantified. The EIS biosensor showed also sensitivity to further cysteine sulphoxides, like methiin, ethiin and propiin, which are common for wild Allium species. Besides the analysis of garlic and garlic-containing preparations, like herbal remedies, this biosensor can be additionally used for breeding research and screening purposes of potential medicinal plants.

High-performance ion-pair chromatography method for simultaneous analysis of alliin, deoxyalliin, allicin and dipeptide precursors in garlic products using multiple mass spectrometry and UV detection

The quality of garlic and garlic products is usually related to their alliin content and allicin release potential. Until now no analytical method was able to quantify simultaneously allicin, its direct precursor alliin ( S-allyl- l-cysteine sulfoxide), SAC ( S-allyl- l-cysteine) as well as various dipeptides that apparently serve as storage compounds in garlic. It is well known that all these intermediates are involved in the allicin biosynthetic pathway. A simple and rapid HPLC method suitable for routine analysis was developed using eluents containing an ion-pairing reagent. Particularly, heptanesulfonate as ion-pairing reagent guarantees a sufficient separation between alliin and the more retained dipeptides at very low pH. Allicin was eluted after 18 min on a 150×3 mm column. Synthetic reference compounds were characterized by the same chromatographic method using a diode-array UV detector and an ion trap mass spectrometer (electrospray ionization) in the multiple MS mode. In routine analysis of garlic bulbs, powders and other products, the diode-array detector is sufficient for a relevant quantification. Our method has been used in studies to improve the quality of garlic and its derived products.

CONTROLLED ATMOSPHERES RETARD SPROUT GROWTH, AFFECT COMPOSITIONAL CHANGES, AND MAINTAIN VISUAL QUALITY ATTRIBUTES OF GARLIC

Garlic storage is important to provide product for fresh market and fresh peeled processing. Garlic bulbs (cv. California Late) and unpeeled cloves were stored in several experiments in air and CA (0.1, 0.5, 1% O2 in combination with 0, 5, 10, 15 and 20% CO2) at 0-1°C for up to 6 months. Quality changes, especially sprout growth, decay and discoloration, were reduced by CO2-containing atmospheres, but the low O2 atmospheres alone generally had little benefit. Storage of garlic with CO2 atmospheres >15% may lead to injury after 4-6 months. Fructan concentrations were similar among air and CA-stored bulbs, but notably decreased under 20% CO2. Thiosulfinate and pyruvate concentrations were generally maintained in CO2-containing atmospheres, but increased in airand low O2stored garlic that permitted sprout development. Alliin concentrations were generally stable during storage up to 6 months, but in some cases decreased significantly under CO2 atmospheres. For fresh peeled garlic, atmospheres of CO2 (5-15%) in air or in low O2 (13%) were effective in retarding discoloration and decay at 5 and 10°C for 3 weeks.

216 Carbohydrate and Alliin Concentrations in Garlic Produced under Different N Fertilzation and Irrigation Regimes

Garlic (cv California Late) was produced under four irrigation regimes (110% and 130% evapotranspiration with two water cut-off dates, 10 and 24 May 1999) in combination with three nitrogen fertilization levels (100, 250, and 400 lb total N). Bulbs were manually harvested mid-June, cured 3 weeks shaded at ambient temperatures and the outer whorl of cloves manually peeled. Samples were freeze-dried, and carbohydrate (fructan and free sugars) and alliin (substrate for alliinase activity and indicator of potential pungency) concentrations were determined by HPLC. The percent dry weight was not affected by the irrigation treatment, but was reduced with increased N rate (41.3% to 39.0%). Alliin concentrations varied from 8.3 to 13.8 mg/g DW for 110% and 130% Eto irrigation treatments. Alliin concentrations were not affected by N fertilization (average = 11.5 mg/g DW). Fructan concentrations were affected by N fertilization treatment, with the highest content (802 mg/g DW) associated with the lowest N level, and the lowest (717 mg/g DW) content in samples from the highest N rate. Sucrose concentrations increased with increased N, but glucose and fructose concentrations did not vary with N fertilization. Fructan as percent of total carbohydrate remained constant across irrigation treatments (96.6% + 0.2%) and across N fertilization treatments (96.6% + 0.3%).

Gas chromatographic determination of S-alk(en)ylcysteine sulfoxides

A new GC method for determination of S-alk(en)ylcysteine sulfoxides, important secondary metabolites occurring in many plant genera, has been developed. The method is based on isolation of the amino acid fraction by ion-exchange chromatography followed by derivatization with ethyl chloroformate at ambient temperature and reduction of derivatized S-alk(en)ylcysteine sulfoxides by sodium iodide. The main advantages of the new method are its high sensitivity, excellent resolution capability, accuracy and reliability, as well as the possibility to identify unknown compounds by means of GC–MS. The content of alliin and other S-alk(en)ylcysteine sulfoxides was determined in nine different samples of garlic ( Allium sativum L.) originating from the Czech Republic, France, and China. The total content of S-alk(en)ylcysteine sulfoxide pool ranged between 0.53 and 1.3% fresh weight, with S-allylcysteine sulfoxide (alliin) being predominant. A novel S-alkylcysteine derivative, S-ethylcysteine sulfoxide (ethiin), not previously reported to occur in Allium species, was found in some of the samples examined.