Effect of cooking on garlic (Allium sativum L.) antiplatelet activity and thiosulfinates content

The raw form of garlic and some of its preparations are widely recognized as antiplatelet agents that may contribute to the prevention of cardiovascular disease. Herein, we examined the in-vitro antiaggregatory activity (IVAA) of human blood platelets induced by extracts of garlic samples that were previously heated (in the form of crushed versus uncrushed cloves) using different cooking methods and intensities. The concentrations of allicin and pyruvate, two predictors of antiplatelet strength, were also monitored. Oven-heating at 200 degrees C or immersing in boiling water for 3 min or less did not affect the ability of garlic to inhibit platelet aggregation (as compared to raw garlic), whereas heating for 6 min completely suppressed IVAA in uncrushed, but not in previously crushed, samples. The latter samples had reduced, yet significant, antiplatelet activity. Prolonged incubation (more than 10 min) at these temperatures completely suppressed IVAA. Microwaved garlic had no effect on platelet aggregation. However, increasing the concentration of garlic juice in the aggregation reaction had a positive IVAA dose response in crushed, but not in uncrushed, microwaved samples. The addition of raw garlic juice to microwaved uncrushed garlic restored a full complement of antiplatelet activity that was completely lost without the garlic addition. Garlic-induced IVAA was always associated with allicin and pyruvate levels. Our results suggest that (1) allicin and thiosulfinates are responsible for the IVAA response, (2) crushing garlic before moderate cooking can reduce the loss of activity, and (3) the partial loss of antithrombotic effect in crushed-cooked garlic may be compensated by increasing the amount consumed.     

Quantitative determination of allicin in garlic: supercritical fluid extraction and standard addition of alliin

A quantitative method is described for the determination of allicin (2-propene-1-sulfinothioic acid S-2-propenyl ester) in garlic, using standard additions of alliin (l-(+)-S-allylcysteine sulfoxide) in conjunction with supercritical fluid extraction (SFE) and high performance liquid chromatography analysis with UV-vis absorbance detection. Optimum CO(2)-SFE conditions provided 96% recovery for allicin with precision of 3% (RSD) for repeat samples. The incorporation of an internal standard (allyl phenyl sulfone) in the SFE step resulted in a modest improvement in recovery (99%) and precision (2% RSD). Standard additions of alliin were converted to allicin in situ by endogenous alliinase (l-(+)-S-alk(en)ylcysteine sulfoxide lyase, EC 4.4.1.4). Complete conversion of the spiked alliin to allicin was achieved by making additions after homogenization-induced conversion of the naturally occurring cysteine sulfoxides to thiosulfinates had taken place, thus eliminating the likelihood of competing reactions. Concentration values for allicin determined in samples of fresh garlic (Allium sativum L. and Allium ampeloprasum) and commercially available garlic powders (Allium sativum L.) by standard addition of alliin were found in all cases to be in statistical agreement (95% confidence interval) with values determined using a secondary allicin standard (concentration determined using published extinction coefficients). This method provides a convenient alternative for assessing the amount of allicin present in fresh and powdered garlic, as alliin is a far more stable and commercially prevalent compound than allicin and is thus more amenable for use as a standard for routine analysis.     

Effect of Processing and Cooking Conditions on Onion ( Allium cepa L.) Induced Antiplatelet Activity and Thiosulfinate Content

Allium vegetables serve as sources of antiplatelet agents that may contribute to the prevention of cardiovascular disease. However, onion and garlic, the major Allium species, are usually cooked before consumption. Here, we examined the effect of cooking on onion in vitro antiplatelet activity (IVAA). Two different cooking systems (convection oven and microwaves) and several time-temperature variables were tested on whole bulbs, quarters of bulbs, and completely crushed bulbs, monitoring the degradation of sulfur antiplatelet compounds (e.g., thiosulfinates) by analysis of pyruvate levels. Although heating was, in general, detrimental for onion IVAA, the extent of this effect varied greatly, from unaffected antiplatelet activity (AA) (i.e., similar to raw onion) to a complete lost of activity, depending upon the manner in which onions were prepared prior to heating, the cooking method, and the intensity of the heat treatment. "Whole", "quarters", and "crushed" onions lost their IVAA after 30, 20, and 10 min of oven heating, respectively. The longer retainment of AA in intact bulbs was attributed to a later alliinase inactivation. Proaggregatory effects observed in samples subjected to the most intense oven and microwave heat treatments suggest that extensively cooked onions may stimulate rather than inhibit platelet aggregation. The efficacy of Allium species as antiplatelet agents, as affected by preparation and cooking conditions, is discussed.     

Identification of two novel pigment precursors and a reddish-purple pigment involved in the blue-green discoloration of onion and garlic

By using a model reaction system representing blue-green discoloration that occurs when purees of onion (Allium cepa L.) and garlic (Allium sativum L.) are mixed, we isolated two pigment precursors (PPs) and a reddish-purple pigment (PUR-1) and determined their chemical structures. PPs were isolated from a heat-treated solution containing color developer (CD) and either l-valine or l-alanine, and their structures were determined as 2-(3,4-dimethylpyrrolyl)-3-methylbutanoic acid (PP-Val), and 2-(3,4-dimethyl-1H-pyrrolyl) propanoic acid (PP-Ala), respectively. Next, PUR-1 was isolated from a heat-treated solution containing PP-Val and allicin, and its structure was determined as (1E)-1-(1-((1S)-1-carboxy-2-methylpropyl)-3,4-dimethyl-1H-pyrrol-2-yl)-prop-1-enylene-3-(1-((1S)-1-carboxy-2-methylpropyl)-3,4-dimethyl-1H-pyrrol-2-ylidenium). The structure of PUR-1 suggested that PP molecules containing a 3,4-dimethyl pyrrole ring had been cross-linked by an allyl group of allicin to form conjugated pigments. While PUR-1 is a dipyrrole compound exhibiting a reddish-purple color, a color shift toward blue to green can be expected as the cross-linking reaction continues to form, for example, tri- or tetrapyrrole compounds.     

Biological and chemical stability of garlic-derived allicin

This study verifies the instability of garlic ( Allium sativum L.)-derived allyl 2-propenylthiosulfinate (allicin) in various aqueous and ethanolic solutions as well as in vegetable oil through chemical and biological analyses performed simultaneously. Crushed fresh garlic cloves generated antibacterial activity and chemically detectable allicin, a major antibacterial principle, and both declined on a daily basis in aqueous and ethanolic solutions at room temperature, showing biological and chemical half-lives of about 6 and 11 days, respectively. Allicin was more stable in 20% alcohol than in water, but surprisingly unstable in vegetable oil, with an activity half-life 0.8 h, as estimated from its antibacterial activity toward Escherichia coli, and a chemical half-life of 3.1 h, based on chromatographic quantification. In alcoholic and aqueous extracts, the biological half-life of allicin tended to be longer than the chemical one, suggesting the occurrence of bioactive compounds other than allicin in the extracts.     

Allicin and allicin-derived garlic compounds increase breath acetone through allyl methyl sulfide: use in measuring allicin bioavailability

Progress in establishing systemic pharmacological effects for fresh, crushed garlic (Allium sativum L) in humans has been hindered by (1) the inability to measure allicin bioavailability, (2) lack of direct evidence that allicin has significant systemic activity at doses of garlic normally consumed, and (3) lack of a model for an acute effect. We have addressed these problems by quantifying the increases in breath acetone and breath allyl methyl sulfide (AMS). The area under the 48 h curve was measured in humans after consumption of standardized garlic preparations, allicin, and allicin-derived compounds, at the equivalent of 7 g of crushed garlic. It was shown that the allyl thiosulfinates (mainly allicin) are solely responsible for breath AMS and increased breath acetone. Diallyl trisulfide, diallyl disulfide, ajoene, and S-allylmercaptocysteine, at isomolar dithioallyl, showed the same quantitative effects as allicin. Consumption of AMS at isomolar allyl also gave the same effects as allicin, indicating that AMS is the main metabolite of allicin and is an active metabolite. In conclusion, allicin and allicin-derived compounds are rapidly metabolized to AMS, a compound which stimulates the production of acetone and which can be used to measure the bioavailability of allicin and, hence, the ability of garlic supplements to represent fresh garlic.     

紫色土增施单质硫对大蒜生长发育和硫素营养的影响

对紫色土施用单质硫条件下大蒜生长发育及硫素营养吸收和代谢进行了研究。结果表明,大蒜株高、叶面积、经济产量(蒜头)和经济系数以中等供硫水平最高,而茎粗和生物产量则以高硫水平最大。大蒜能耐高浓度的硫素供应,在0～120kghm-2供硫情况下,其全硫(TS)、水溶性硫(SS)和无机硫(Io-S)含量均随供硫量的增加而上升,小分子水溶性含硫氨基酸含量(Ws-S)以低硫水平最高,而大分子蛋白质硫含量(Wis-S)以中等供硫水平时最高,与大蒜素含量的变化一致。大蒜素含量与不同硫组分比率的关系分析发现,大蒜素含量与Ws-S/TS呈极显著的正相关(r=0.752 ),与Io-S/SS呈显著的正相关(r=0.702 )。全氮含量与全硫含量变化趋势一致,全磷含量以低硫水平时最高,全钾含量以中等供硫水平时最高。     

Inhibitory effect of α-lipoic acid on platelet aggregation is mediated by PPARs

Peroxisome proliferator-activated receptors (PPARs) isoforms (α, β/δ, and γ are present in human platelets, and activation of PPARs inhibits platelet aggregation. α-Lipoic acid (ALA), occurring naturally in human food, has been reported to exhibit an antiplatelet activity. However, the mechanisms underlying ALA-mediated inhibition of platelet aggregation remain unknown. The aim of this study was to investigate whether the antiplatelet activity of ALA is mediated by PPARs. ALA itself significantly induced PPARα/γ activation in platelets and increased intracellular amounts of PPARα/γ by blocking PPARα/γ secretion from arachidonic acid (AA)-activated platelets. Moreover, ALA significantly inhibited AA-induced platelet aggregation, Ca(2+) mobilization, and cyclooxygenase-1 (COX-1) activity, but increased cyclic AMP production in rabbit washed platelets. Importantly, ALA also enhanced interaction of PPARα/γ with protein kinase Cα (PKCα) and COX-1 accompanied by an inhibition of PKCα activity in resting and AA-activated platelets. However, the above effects of ALA on platelets were markedly reversed by simultaneous addition of selective PPARα antagonist (GW6471) or PPARγ antagonist (GW9662). Taken together, the present study provides a novel mechanism by which ALA inhibition of platelet aggregation is mediated by PPARα/γ-dependent processes, which involve interaction with PKCα and COX-1, increase of cyclic AMP formation, and inhibition of intracellular Ca(2+) mobilization.     

Apigenin inhibits platelet adhesion and thrombus formation and synergizes with aspirin in the suppression of the arachidonic acid pathway

Previous studies using washed platelets demonstrated that certain flavonoids inhibit platelet function through several mechanisms including blockade of TxA(2) receptors (TPs). We aimed to analyze the binding capacity of flavonoids to TPs in platelet rich plasma (PRP), investigated their effect in flowing blood, and evaluated the ability of apigenin to improve the efficacy of aspirin in the inhibition of platelet aggregation. The binding of flavonoids to TPs in PRP was explored using binding assays and the TP antagonist [ (3)H]SQ29548. Effects of flavonoids on platelet adhesion were assessed using arterial subendothelium with annular plate perfusion chambers, and global evaluation of apigenin on high-shear-dependent platelet function was determined by the PFA-100. To evaluate the ability of apigenin to potentiate the effect of aspirin, arachidonic acid-induced platelet aggregation was measured prior to and after consumption of subaggregatory doses of aspirin in the presence or absence of apigenin. Binding assays revealed that apigenin was an efficient competitor of [ (3)H]SQ29548 binding to PRP ( K i = 155.3 +/- 65.4 microM), and perfusion studies showed that apigenin, genistein, and catechin significantly diminished thrombus formation when compared to control (26.2 +/- 3.8, 33.1 +/- 5.2, and 26.2 +/- 5.2 vs 76.6 +/- 2.6%, respectively; p < 0.05). Apigenin, similarly to the TP antagonist SQ29548, significantly prolonged collagen epinephrine-induced PFA-100 closure time in comparison to the control and, when added to platelets that had been exposed in vivo to aspirin, potentiated its inhibitory effect on platelet aggregation. The inhibitory effect of some flavonoids in the presence of plasma, particularly apigenin, might in part rely on TxA(2) receptor antagonism. There is a clear increase in the ex vivo antiplatelet effect of aspirin in the presence of apigenin, which encourages the idea of the combined use of aspirin and certain flavonoids in patients in which aspirin fails to properly suppress the TxA(2) pathway.     

Antiplatelet activity of soy sauce as functional seasoning.

In seeking the functionality of foodstuffs applicable to medicine, soy sauce was found to show antiplatelet activity. Therefore, the active components in soy sauce were purified, structurally identified, and studied for their inhibitory effects on the aggregation of human platelets. Aqueous 2-fold diluents of soy sauce inhibited platelet aggregation induced by collagen and epinephrine depending on the dilution factor. Since a basic extract with diethyl ether completely inhibited collagen-induced aggregation, it was subjected to serial extractions and multistep HPLC fractionations for purifying antiplatelet components. The finally obtained isolates were identified as 1-methyl-1,2,3,4-tetrahydro-beta-carboline and 1-methyl-beta-carboline on the basis of EI-MS, (1)H NMR, diode array, and fluorescence spectra. Their spectral data and chromatographic behaviors were the same as those of synthetic ones. 1-Methyl-1,2,3, 4-tetrahydro-beta-carboline showed mean concentrations (n = 5-6) of 4.6, 4.2, 28.6, 11.6, and 65.8 microgram/mL to produce 50% inhibition of the maximal aggregation response induced by epinephrine, platelet-activating factor, collagen, adenosine 5'-diphosphate, and thrombin, respectively. Its inhibitory effect was much greater than that of 1-methyl-beta-carboline on platelet aggregation by all the tested inducers. The quantitative HPLC analysis revealed that the significant amounts of both antiplatelet compounds were uniformly contained in commercially available soy sauce. From these results, soy sauce may be referred to as functional seasoning containing alkaloidal components with the potent preventive effect on thrombus formation.     

硒对大蒜生理特性、含硒量及品质的影响

以金蒜3号为试材,采用盆栽试验,通过叶面喷施不同浓度、不同次数的亚硒酸钠研究了硒对大蒜生理特性、含硒量和品质的影响。结果表明:在生长期4月5日、15日和25日累计喷施1次、2次、3次3个浓度（5、10、15 mg/L）的亚硒酸钠溶液,均不同程度影响大蒜生理特性、含硒量和品质。与对照相比,叶面喷施2次10 mg/L的亚硒酸钠溶液,可显著提高叶片中光合色素含量和光合性能;能显著改善蒜薹和鳞茎品质,单薹鲜质量和单头鲜质量分别提高68.25%和29.00%,可溶性糖、可溶性蛋白、维生素C含量分别提高73.05%、104.66%、18.95%和82.01%、51.27%、69.82%;并可有效提高鳞茎中游离氨基酸含量（38.91%）,降低蒜薹中游离氨基酸含量（36.95%）;同时可降低蒜薹和鳞茎中大蒜素含量,但差异不显著。蒜薹和鳞茎硒含量随硒浓度和喷施次数的增加而显著提高,最高可达19.81和23.96 mg/kg,为对照的3.85和4.41倍。综合各因素指标,大蒜以叶面喷施2次10 mg/L的亚硒酸钠为宜。     

适宜的氮硫配施提高基质栽培大蒜鳞茎品质

试验采用蛭石-珍珠岩盆栽方式,研究了不同浓度氮(5、10、20 mmol/L,记作N1、N2、N3)、硫(2、4、8 mmol/L,记作S1、S2、S3)配施条件对大蒜鳞茎品质的影响,并采用隶属函数法对大蒜品质进行综合评价,分析鳞茎品质与氮硫互作水平的响应关系。试验结果表明,氮素、硫素对大蒜主要营养成分含量影响不尽相同,氮硫配施能够不同程度提高大蒜鳞茎中可溶性蛋白、VC、大蒜多糖、游离氨基酸的含量,且氮、硫单一因素对大蒜鳞茎品质影响远低于元素间交互作用。但在氮素适宜水平下(10~20 mmol/L),硫素水平对大蒜鳞茎中大蒜辣素含量有显著影响,提高硫浓度能够显著增加大蒜辣素含量,而在硫素浓度过高时(8 mmol/L)则明显抑制该物质合成。隶属函数法综合评价以N3S2处理组大蒜鳞茎品质最优,隶属函数值为0.81,对照组(CK)最差。综合分析可见,无机基质栽培条件下氮硫配施显著影响大蒜鳞茎品质,氮素浓度20 mmol/L(N3)、硫素浓度4 mmol/L(S2)时最利于大蒜鳞茎品质的提升,为大蒜无机基质栽培最佳氮、硫配施水平,且两元素配施存在明显的互作效应。该结论为大蒜栽培氮、硫元素合理配施提供试验指导。     

Isolation and identification of antiplatelet aggregatory principles from the leaves of Piper lolot

The methanolic extract of Piper lolot, having shown potent inhibitory activity on platelet aggregation induced by arachidonic acid (AA) and platelet activating factor (PAF), was subjected to activity-guided isolation to yield twelve new amide alkaloids, piperlotine A-L (1-12), along with twenty-nine known compounds. Their structures were elucidated on the basis of spectroscopic analysis. The isolated compounds were tested for their inhibitory activity on the rabbit platelet aggregation. The compounds piperlotine A (1), piperlotine C (3), piperlotine D (4), piperlotine E (5), 3-phenyl-1-(2,4,6-trihydroxyphenyl)propan-1-one (21), 3-(4-methoxyphenyl)-1-(2,4,6-trihydroxyphenyl)propan-1-one (22), 1-trans-cinnamoylpyrrolidine (24), sarmentine (26), pellitorine (27), methyl 3-phenylpropionate (32), and (10S)-10-hydroxypheophorbide a methyl ester (40) showed potent antiplatelet aggregation activity.     

Chemical and genomic combined approach applied to the characterization and identification of Italian Allium sativum L

Chemotype analyses and random amplified polymorphic DNA (RAPD) genomic analyses have been applied to the characterization of Allium sativum variety from Voghiera (Ferrara, Italy), a typical Italian product actually demanding the Protected Designation of Origin (PDO). The garlic from Voghiera is characterized by peculiar morphological and composition characteristics. The proximate composition and atomic absorbance spectrometry elemental pattern of this garlic suggested as the chemical composition did not depend on the intrinsic pedologic soil features only, but it was probably connected to some peculiar genetic characters. Amplification of genomic DNA using random primers highlighted a good clustering differentiating of Voghiera Allium sativum from five commercial reference samples used in this study (Piacentino, Serena, France, China, and Adriano varieties), confirming the existence of intervarietal genetic difference. The intravarietal polymorphisms of Voghiera samples were low.     

超高压和超声波预处理对蒜片热风干燥过程及品质的影响

为缩短蒜片热风干燥时间,提高干制蒜片品质,将超高压与超声波技术应用到蒜片干燥前处理,并利用低场核磁共振技术(Low Field Nuclear Magnetic Resonance, LF-NMR)分析预处理对蒜片干燥过程中内部不同状态水分迁移的影响。结果表明:干燥前超声或超高压预处理有利于加快干燥过程,超声、超高压和超高压-超声预处理的干燥时间分别比没有预处理的对照组降低了37.50%、43.75%、62.50%;三种预处理均可以降低干燥能耗,尤以超高压-超声最为显著(P0.05),相比对照降低了32.31%。低场核磁共振测定结果表明,预处理后蒜片中存在三种状态的水分,自由水占的比例最大,不易流动水次之,结合水最少。干燥过程中脱除的主要是自由水,超声和超高压均降低了自由水的脱除时间,超高压-超声预处理脱除自由水的时间最少,为90 min,说明超高压和超声联合更有利于减弱蒜片组织对水分的束缚并增强水分的流动性。通过扫描电镜发现,超高压使蒜片细胞间隙增大,细胞壁破坏严重,而经超声预处理的蒜片组织出现疏松多孔的结构,超高压-超声扩增了蒜片的显微通道,验证了超高压-超声更有利于提高水分的流动性。经超高压或超声预处理后,干制蒜片的色差降低,复水比和蒜素含量显著提高(P0.05),超高压与超声联合预处理得到的干制蒜片品质较优,色差值为16.11,大蒜素含量为2.67 mg/g,复水比为2.90 g/g。研究结果为高效节能、高品质的大蒜干燥技术提供理论参考。     

C-phycocyanin, a very potent and novel platelet aggregation inhibitor from Spirulina platensis

The aim of this study was to systematically examine the inhibitory mechanisms of C-phycocyanin (C-PC), one of the major phycobiliproteins of Spirulina platensis (a blue-green alga), in platelet activation. In this study, C-PC concentration-dependently (0.5-10 nM) inhibited platelet aggregation stimulated by agonists. C-PC (4 and 8 nM) inhibited intracellular Ca2+ mobilization and thromboxane A2 formation but not phosphoinositide breakdown stimulated by collagen (1 microg/mL) in human platelets. In addition, C-PC (4 and 8 nM) markedly increased levels of cyclic GMP and cyclic GMP-induced vasodilator-stimulated phosphoprotein (VASP) Ser(157) phosphorylation. Rapid phosphorylation of a platelet protein of Mw 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12,13-dibutyrate (150 nM). This phosphorylation was markedly inhibited by C-PC (4 and 8 nM). In addition, C-PC (4 and 8 nM) markedly reduced the electron spin resonance (ESR) signal intensity of hydroxyl radicals in collagen (1 microg/mL)-activated platelets. The present study reports on a novel and very potent (in nanomolar concentrations) antiplatelet agent, C-PC, which is involved in the following inhibitory pathways: (1) C-phycocyanin increases cyclic GMP/VASP Ser157 phosphorylation and subsequently inhibits protein kinase C activity, resulting in inhibition of both P47 phosphorylation and intracellular Ca2+ mobilization, and (2) C-PC may inhibit free radicals (such as hydroxyl radicals) released from activated platelets, which ultimately inhibits platelet aggregation. These results strongly indicate that C-PC appears to represent a novel and potential antiplatelet agent for treatment of arterial thromboembolism.     

Inhibiting effects of egg white dry-heated at 120 degrees C on heat aggregation and coagulation of egg white and characteristics of dry-heated egg white.

Dialyzed and freeze-dried egg white (FDEW) was dry-heated at 120 degrees C for up to 6 h. The inhibiting effects of the dry-heated egg white (DHEW) on the heat aggregation and coagulation of egg white (as 10% FDEW solution) and characteristics of the DHEW were examined. From the changes in turbidities and soluble protein contents of supernatant in various mixtures of 10% FDEW and DHEW solutions induced by heating (60 degrees C, 5 min), it was found that the inhibiting capacity increased with increases in the dry-heating time (DHT). The FDEW proteins were denatured with a mild conformational change (not secondary but tertiary structure) with the increase in DHT and aggregated partially. However, the more transparent solutions of DHEW containing soluble aggregates according to DHT were also obtained after heating. The transparency according to DHT came to be scarcely affected by the NaCl concentration and the dilution with diluents containing SDS, urea, and 2-mercaptoethanol. These findings suggest that the heat aggregations and coagulations of ovotransferrin and lysozyme in the FDEW were inhibited by their bindings with the soluble aggregates in DHEW.     

Exploration of the antiplatelet activity profile of betulinic acid on human platelets

Betulinic acid, a natural pentacyclic triterpene acid, presents a diverse mode of biological actions including antiretroviral, antibacterial, antimalarial, and anti-inflammatory activities. The potency of betulinic acid as an inhibitor of human platelet activation was evaluated, and its antiplatelet profile against in vitro platelet aggregation, induced by several platelet agonists (adenosine diphosphate, thrombin receptor activator peptide-14, and arachidonic acid), was explored. Flow cytometric analysis was performed to examine the effect of betulinic acid on P-selectin membrane expression and PAC-1 binding to activated platelets. Betulinic acid potently inhibits platelet aggregation and also reduced PAC-1 binding and the membrane expression of P-selectin. Principal component analysis was used to screen, on the chemical property space, for potential common pharmacophores of betulinic acid with approved antithrombotic drugs. A common pharmacophore was defined between the NMR-derived structure of betulinic acid and prostacyclin agonists (PGI2), and the importance of its carboxylate group in its antiplatelet activity was determined. The present results indicate that betulinic acid has potential use as an antithrombotic compound and suggest that the mechanism underlying the antiplatelet effects of betulinic acid is similar to that of the PGI2 receptor agonists, a hypothesis that deserves further investigation.     

Effects of benomyl and captan on rhizosphere fungi and the growth of Allium cepa

The effects of soil treatment with Benomyl or Captan on the rhizosphere fungi of Allium cepa were investigated. The mycoflora of the rhizosphere was considerably inhibited by both fungicides. Captan was more inhibiting than Benomyl, but the latter seemed to be more persistent. The fungi isolated from the rhizosphere of control plants, plants treated with Benomyl or with Captan are listed and differences between treatments are discussed. The total number of species isolated from the rhizosphere of control plants was 65, Benomyl 44 and Captan 29. The growth of onions receiving fungicide treatment was greatly reduced: differences in dry weight were 31% (Captan) and 36% (Benomyl) less than control plants.     

Low allicin release from garlic supplements: a major problem due to the sensitivities of alliinase activity

Most garlic supplements are standardized on allicin potential and are enteric-coated to prevent gastric acid inactivation of the allicin-producing enzyme, alliinase. To determine whether these products release the claimed amount of allicin under simulated gastrointestinal conditions, USP dissolution method 724A for drug release was applied to all 24 known brands of enteric-coated tablets. It was found that nearly all brands employed effective coatings and that they met their claims for allicin potential when crushed and suspended in water. However, all brands except one gave low dissolution allicin release, with 83% of the brands releasing less than 15% of their potential. The low allicin release was found to be due to both impaired alliinase activity, mostly caused by tablet excipients, and to slow tablet disintegration, which also impairs alliinase activity. Only when tablets had high alliinase activity and disintegrated rapidly did they show high allicin release. The ability of USP 724A to estimate allicin release in vivo was validated by monitoring breath levels of the allicin metabolite, allyl methyl sulfide. In conclusion, garlic powder supplements should no longer be standardized on allicin potential, but rather on dissolution allicin release.