Reversible and covalent binding of 5-(hydroxymethyl)-2-furaldehyde (HMF) with lysine and selected amino acids

The chemical reactivity of 5-(hydroxymethyl)-2-furaldehyde (HMF) with lysine, glycine, and proline was studied using isotope labeling technique. To confirm the formation of HMF adducts in glucose amino acid model systems, a useful strategy was developed in which products simultaneously possessing six glucose (HMF moiety) and any number of amino acid carbon atoms in addition to nitrogen were targeted using specifically labeled precursors such as [(15)N(α)]lysine·2HCl, [(15)N(ε)]lysine·2HCl, [U-(13)C(6)]lysine·2HCl, [(13)C(6)]lysine·2HCl, and [U-(13)C(6)]glucose in the case of lysine model system. In addition, model systems containing HMF and amino acids were also studied to confirm specific adduct formation. Complete labeling studies along with structural analysis using appropriate synthetic precursors such as HMF Schiff base adducts of piperidine and glycine have indicated that HMF generated in the glucose/amino acid model systems initially forms a Schiff base adduct that can undergo decarboxylation through an oxazolidin-5-one intermediate and form two isomeric decarboxylated Schiff bases. Unlike the Schiff bases resulting from primary amines or amino acids such as glycine or lysine, those resulting from secondary amino acids such as proline or secondary amines such as piperidine can further undergo vinylogous Amadori rearrangement, forming N-substituted 5-(aminomethyl)furan-2-carbaldehyde derivatives.     

Isotope labeling studies on the formation of 5-(hydroxymethyl)-2-furaldehyde (HMF) from sucrose by pyrolysis-GC/MS

Although it is generally assumed that the reactivity of sucrose, a nonreducing sugar, in the Maillard reaction is due to its hydrolysis into free glucose and fructose, however, no direct evidence has been provided for this pathway, especially in dry and high temperature systems. Using specifically (13)C-labeled sucrose at C-1 of the fructose moiety, HMF formation was studied at different temperatures. Under dry pyrolytic conditions and at temperatures above 250 degrees C, 90% of HMF originated from fructose moiety and only 10% originated from glucose. Alternatively, when sucrose was refluxed in acidic methanol at 65 degrees C, 100% of HMF was generated from the glucose moiety. Moreover, the relative efficiency of the known HMF precursor 3-deoxyglucosone to generate HMF was compared to that of glucose, fructose and sucrose. Glucose exhibited a much lower conversion rate than 3-deoxyglucosone, however, both fructose and sucrose showed much higher conversion rates than 3-deoxyglucosone thus precluding it as a major precursor of HMF in fructose and sucrose solutions. Based on the data generated, a mechanism of HMF formation from sucrose is proposed. According to this proposal sucrose degrades into glucose and a very reactive fructofuranosyl cation. In dry systems this cation can be effectively converted directly into HMF.     

Thermal decomposition of 5-(hydroxymethyl)-2-furaldehyde (HMF) and its further transformations in the presence of glycine

Thermal decomposition of HMF has been so far studied indirectly through carbohydrate degradation reactions assuming HMF as the main product. Such studies, however, do not necessarily generate relevant information on HMF decomposition because many other products are generated simultaneously. Direct thermal decomposition using different concentrations of HMF in silica gel was studied using pyrolysis-GC-MS. Undiluted HMF generated four peaks corresponding to 5-methylfurfural, 2,5-furandicarboxaldehdye, HMF, and a major unknown peak at retention time of 20.73 min. The diluted HMF in silica gel (15-fold) generated only the first three peaks. The generation of the unknown peak was dependent on the concentration of HMF, indicating the possibility of a dimeric structure; furthermore, when HMF was generated from [U-13C6]glucose in the reaction mixture, the highest mass in the spectrum of the unknown peak showed the incorporation of 11 carbon atoms from the glucose. Thermal decomposition studies of HMF have also indicated that in the absence of amino acids it can mainly dimerize and the initially formed dimer can degrade to generate 5-methylfurfural and 2,5-furandicarboxaldehyde. On the other hand, thermal degradation of HMF in the presence of glycine generated Schiff base adducts of HMF, 5-methylfurfural, and 2,5-furandicarboxaldehdye in addition to 2-acetyl-5-methylfuran and a newly discovered adduct, 5-[(dimethylamino)methyl]-2-furanmethanol.     

Investigation of the reaction between 4-hydroxy-5-methyl-3(2H)-furanone and cysteine or hydrogen sulfide at pH 4.5

Reaction of 4-hydroxy-5-methyl-3(2H)-furanone (HMF) with cysteine or hydrogen sulfide at pH 4.5 for 60 min at 140 degrees C produced complex mixtures of volatile compounds, the majority of which contained sulfur. Sixty-nine compounds were identified, some tentatively, by GC/MS. These included disulfides (26), thiols (7), dithiolanones (6), thiophenones (4), dithianones (3), and thienothiophenes (6). The main non-sulfur compounds were 2, 3-pentanedione, 2,4-pentanedione, and 3,4-hexanedione. Both systems produced approximately the same total quantity of volatile compounds, but the reaction containing cysteine gave the larger number of individual compounds, with thiols quantitatively the dominant components. By comparison, the major products formed in the reaction with hydrogen sulfide were the dithiolanones. Reaction pathways are presented for the major products and, where applicable, possible reasons for the differences in composition of the two systems are discussed. The contribution of these reactions, and their products, to the flavor of roasted foods is considered.     

Heterocyclic volatiles formed by heating cysteine or hydrogen sulfide with 4-hydroxy-5-methyl-3(2H)-furanone at pH 6.5

The reaction of 4-hydroxy-5-methyl-3(2H)-furanone (HMF) with cysteine or hydrogen sulfide at pH 6.5 for 60 min at 140 degrees C produced complex mixtures of volatile compounds, the majority of these containing either sulfur or nitrogen. Of the 68 compounds detected, 63 were identified, some tentatively, by GC-MS. Among the identified compounds were thiophenes (10), thiophenones (6), thienothiophenes (5), thiazoles (5), trithiolanes (4), pyrazines (6), and oxazoles (4). More compounds were produced in the reaction of HMF with cysteine (63) than were formed in the reaction with hydrogen sulfide (33). In both systems, thiophenones were major reaction products, accounting for 25-36% of the total volatiles formed. Possible reasons for the differences in the composition of the two systems are discussed. The contributions of these reactions, and their products, to the flavor of heated foods are considered.     

Simulated acid rain (H2SO4) and microbial activity in soil

Soil mixtures containing 9% kaolinite, 9% montmorillonite, or no clay supplements were amended with 1% glucose and treated with H₂SO₄ to lower their bulk pH to levels ranging from 5.4 to 0.8. Acidification had little effect on soil respiration (CO₂ evolution) until the pH was lowered below 3. Glucose was not degraded at approximately pH 2 but was degraded once the soil pH was raised to non-inhibitory levels, i.e. pH 4.1–4.3. When the soil pH was reduced to 1.4 or below, it was necessary to reinoculate the soil and raise the pH to a non-inhibitory level to obtain CO₂ evolution. The addition of clay minerals, particularly montmorillonite, mitigated the toxic effect of H₂SO₄, especially at pH values below 3. The growth of Aspergillus niger, A. flavipes, Trichoderma viride and Penicillium brefeldianum was reduced or completely inhibited in soils acidified below pH 3.5. The addition of montmorillonite enhanced fungal growth under these acidic conditions, but kaolinite had no effect.     

Simultaneous quantitation of 2-acetyl-4-tetrahydroxybutylimidazole, 2- and 4-methylimidazoles, and 5-hydroxymethylfurfural in beverages by ultrahigh-performance liquid chromatography-tandem mass spectrometry

An ultrahigh-performance liquid chromatography (UHPLC) tandem mass spectrometric (MS/MS) method was developed for the simultaneous quantification of 2-acetyl-4-tetrahydroxybutylimidazole (THI), 2- and 4-methylimidazoles (2-MI and 4-MI), and 5-hydroxymethylfurfural (HMF) in beverage samples. A C30 reversed-phase column was used in this method, providing sufficient retention and total resolution for all targeted analytes, with an MS/MS instrument operated in selected reaction monitoring (SRM) mode for sensitive and selective detection using isotope-labeled 4-methyl-d(3)-imidazole (4-MI-d(3)) as the internal standard (IS). This method demonstrates lower limit of quantification (LLOQ) at 1 ng/mL and coefficient of determination (r(2)) >0.999 for each analyte with a calibration range established from 1 to 500 ng/mL. This method also demonstrates excellent quantification accuracy (84.6-105% at 5 ng/mL, n = 7), precision (RSD < 7% at 5 ng/mL, n = 7), and recovery (88.8-99.5% at 10, 100, and 200 ng/mL, n = 3). Seventeen carbonated beverage samples were tested (n = 2) in this study including 13 dark-colored beverage samples with different flavors and varieties and 4 light-colored beverage samples. Three target analytes were quantified in these samples with concentrations in the range from 284 to 644 ng/mL for 4-MI and from 706 to 4940 ng/mL for HMF. THI was detected in only one sample at 6.35 ng/mL.     

Identification and quantitation of key aroma compounds formed in Maillard-type reactions of fructose with cysteamine or isothiaproline (1,3-thiazolidine-2-carboxylic acid)

Fructose was reacted in the presence of either cysteamine (model A) or isothiaproline (model B) in aqueous buffer at 145 degrees C and pH 7.0. Application of an aroma extract dilution analysis on the bulk of the volatile compounds formed in model A revealed 5-acetyl-3,4-dihydro-2H-1,4-thiazine (19), N-(2-mercaptoethyl)-1,3-thiazolidine (16), 4-hydroxy-2,5-dimethyl-3(2H)-furanone (15), and 2-acetyl-2-thiazoline (11) as the key aroma compounds among the 10 odorants detected. A similar set of aroma compounds was formed when isothiaproline was reacted (model B), but the flavor dilution factors were generally lower. Substitution of the buffer by silica gel/water (9 + 1 w/w) in both models and application of 150 degrees C for 10 min also gave the same key odorants from both thio compounds; however, under these conditions isothiaproline was the better precursor of, in particular, 19 and 11. Quantitative measurements performed by means of stable isotope dilution assays revealed a significant effect of the pH on odorant formation. For example, in model A, formation of 19 as well as of 11 was suppressed at pH values <5.0. A clear maximum was, however, found for 19 at pH 7.0 (approximately 1 mol % yield), whereas 11 increased with increasing pH from 7.0 to 9.0.     

Formation of carcinogenic 4(5)-methylimidazole in Maillard reaction systems

4(5)-Methylimidazole has received the attention of federal and state regulatory agencies because of its carcinogenicity and common presence in foods and beverages. In the present study, the formation of 4(5)-methylimidazole in Maillard reaction model systems consisting of D-glucose/NH(3), L-rhamnose/NH(3), methylglyoxal/NH(3), and methylglyoxal/formaldehyde/NH(3) was investigated. 4(5)-Methylimidazole was formed at levels ranging from 0.49 to 0.71 mg/mL in the d-glucose/NH(3) model system. The formation of 4(5)-methylimidazole was slightly higher in the L-rhamnose/NH(3) system (0.91 mg/mL) than in the d-glucose/NH(3) system (0.71 mg/mL) under the conditions used in the present study. A methylglyoxal/NH(3) system produced significantly higher levels of 4(5)-methylimidazole (5.70 mg/mL), suggesting that methylglyoxal is an important precursor of 4(5)-methylimidazole. Ammonolysis of methylglyoxal, which is one of the glucose degradation products, was proposed to form formamide, which subsequently reacted with 2-aminopropanal (α-aminocarbonyl intermediate) formed from methylglyoxal to give 4- or 5-methylimidazole. The levels of 4(5)-methylimidazole found in commercial cola soft drinks range from 0.30 μg/mL (brand 3) to 0.36 μg/mL (brands 1 and 5).     

Analysis of a model reaction system containing cysteine and (E)-2-methyl-2-butenal, (E)-2-hexenal, or mesityl oxide

Cysteine conjugates, resulting from the addition of cysteine to alpha,beta-unsaturated carbonyl compounds, are important precursors of odorant sulfur compounds in food flavors. The aim of this work was to better understand this chemistry in the light of the unexpected double addition of cysteine to two unsaturated aldehydes. These reactions were studied as a function of pH. When (E)-2-methyl-2-butenal (tiglic aldehyde, 4) was treated with cysteine in water at pH 8, the major product formed was the new compound (4R)-2-(2-[[(2R)-2-amino-2-carboxyethyl]thio]methylpropyl)-1,3-thiazolidine-4-carboxylic acid (6). Under acidic conditions (pH 1), we also observed a double addition, but the second cysteine was linked by a vinylic sulfide bond to form the previously unreported major product, (2R,2'R,E)-S,S'-(2,3-dimethyl-1-propene-1,3-diyl)bis-cysteine (7). When (E)-2-hexenal (12) was treated with cysteine under acidic conditions, the major product was the novel (4R,2' 'R)-2-[2'-(2' '-amino-2' '-carboxyethylthio)pentyl]-1,3-thiazolidine-4-carboxylic acid (13), and the formation of an vinylic sulfide compound analogous to 7 was not observed. Reduction of the acidic crude reaction mixture with NaBH(4) afforded 13 and the cysteine derivative (R)-S-[1-(2-hydroxyethyl)butyl]cysteine (14) in 14% yield. Treating (E)-2-hexenal with cysteine at pH 8 followed by NaBH(4) reduction yielded the new product (3R)-7-propylhexahydro-1,4-thiazepine-3-carboxylic acid (15). Addition of cysteine to mesityl oxide (16), at pH 8, followed by reduction with NaBH(4) furnished (R)-S-(3-hydroxy-1,1-dimethylbutyl)cysteine (3) and the new compound (3R)-hexahydro-5,7,7-trimethyl-1,4-thiazepine-3-carboxylic acid (18).     

Degradation of the Amadori compound N-(1-deoxy-D-fructos-1-yl)glycine in aqueous model systems

The fate of the Amadori compound N-(1-deoxy-D-fructos-1-yl)glycine (DFG) was studied in aqueous model systems as a function of time and pH. The samples were reacted at 90 degrees C for up to 7 h while maintaining the pH constant at 5, 6, 7, or 8. Special attention was paid to the effect of phosphate on the formation of glycine and the parent sugars glucose and mannose, as well as formic and acetic acid. These compounds and DFG were quantified by high-performance anion-exchange chromatography. The rate of DFG degradation increased with pH. Addition of phosphate accelerated this reaction, particularly at pH 5-7. The rate of glycine formation increased with pH in both the absence and presence of phosphate. High glycine concentrations (60-70 mol %) were obtained, preferably at pH 6-8 with phosphate. However, the yield of glycine formed from DFG decreased at the advanced reaction stage for all pH values studied, both in water and in phosphate buffer. The rate of parent sugar formation increased from pH 5 to pH 7 in the absence of phosphate, leading to glucose and mannose in a constant ratio of 7:3. Addition of phosphate accelerated this reaction, yielding up to 18% parent sugars, most likely formed by reverse Amadori rearrangement. The formation rate of acetic and formic acid increased with increasing pH. The sum of both acids attained 76 mol %. However, the acetic acid concentrations were much higher than those of formic acid.     

Reactivity of epicatechin in aqueous glycine and glucose maillard reaction models: quenching of C2, C3, and C4 sugar fragments

Mechanisms of how epicatechin alters the pathways of the Maillard reaction were investigated. Carbon-13 and nitrogen-15 labeling studies were utilized to define the reactivity of epicatechin with glucose, glycine, and/or reaction products in an aqueous model (pH 7, 125 degrees C for 30 min) via GC, GC/MS and HPLC/MS analysis. Quantification of the volatile reaction product isotopomers by GC/MS from a 1:1 labeled to unlabeled glucose (carbohydrate module labeling technique) plus glycine model system indicated the formation of 2,3-butanedione and acetol were primarily formed via intact C4 and C3 sugar fragments, whereas pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine, and cyclotene were primarily formed via intact C2/C2, C2/C3, C3/C3, C3/C3, and C3/C3 sugar fragment pairs, respectively. The formation of these seven compounds was also reported by GC analysis to be dramatically inhibited when epicatechin was added to the glucose/glycine model system (observed 9-113-fold reduction). HPLC/MS analysis of both the glucose-labeled and glycine-labeled model systems with and without epicatechin indicated that epicatechin reacted directly with C2, C3, and C4 sugar fragments, while epicatechin did not report any direct reactivity with glycine. In conclusion, the quenching of sugar fragmentation products via epicatechin was correlated with the observed inhibition on volatile compound formation when epicatechin was added to a glucose/glycine aqueous reaction model system.     

Formation of 5-methyl-4-hydroxy-3[2H]-furanone in cytosolic extracts obtained from Zygosaccharomyces rouxii

Formation of the flavor compound and precursor 4-hydroxy-5-methyl-3[2H]-furanone (HMF, norfuraneol) was demonstrated in cytosolic protein extracts obtained from Zygosaccharomyces rouxii after incubation with a number of carbohydrate phosphates. 4-Hydroxy-5-methyl-3[2H]-furanone was produced from d-fructose-1,6-diphosphate, d-fructose-6-phosphate, d-glucose-6-phosphate, 6-phosphogluconate, d-ribose-5-phosphate, and d-ribulose-1,5-diphosphate. Enzyme assays revealed d-fructose-1,6-diphosphatase, phosphohexose isomerase, d-glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase activity in the cytosolic extracts. Model studies showed the spontaneous formation of HMF from d-ribulose-5-phosphate. It is assumed that d-ribulose-5-phosphate is generated in cytosolic extracts by the action of the investigated enzymes from the carbohydrate phosphates and is then chemically transformed to HMF. The hypothesis was proven by the production of HMF in solutions containing commercially available enzymes and [6-(13)C]-d-glucose-6-phosphate.     

生物基呋喃与甲醇耦合催化热解制备芳烃化合物

为了提高芳烃的选择性产率和减少催化剂的积碳,以生物基呋喃为原料,以甲醇为耦合试剂进行催化共热解,探讨工艺条件对芳烃的选择性产率的影响,同时对其转化路径以及催化剂的积碳进行分析。结果表明:呋喃与甲醇耦合协同催化共热解可以有效的提高芳烃的选择性产率,二者具有明显的协同效应,甲醇的添加促进了甲醇制烯烃(methanol to olefin,MTO)反应、Diels-Alder环加成反应以及苯烷基化反应的发生,高温促使多烷基化合物以及萘及其同系物选择性产率的增加;强酸性促进了甲醇的脱水以及Diels-Alder环加成反应;同时,羰基抑制了呋喃环和烯烃的Diels-Alder反应,而羟基的存在有效的促进了甲苯以及二甲苯的生成,因此,当采用HZSM-5(SiO2/Al2O3=25)为催化剂,当热解温度为500℃,催化温度为550℃,MF∶甲醇=1∶5,物质的进样量为0.2 mL/min时,其芳烃的选择性产率达到99.73%,积碳量达到11.06%,苯、甲苯、二甲苯以及乙苯的总含量达到40.49%,萘及其同系物的含量仅为10.15%,有效的提高了烷基苯的选择性产率。     

Role of different factors affecting the formation of 5-hydroxymethyl-2-furancarboxaldehyde in heated grape must

The influence of concentration and water activity (a(w)) on the formation of 5-hydroxymethyl-2-furancarboxaldehyde (HMF) in thermally treated grape must was evaluated. Must was cryoconcentrated and then heated to study the pure effect of sugar concentration. Moreover, NaCl was added to the must to lower a(w), maintaining the same sugar concentration, with the purpose of evaluating the pure effect of a(w). Finally, the influence of minimal pH changes on the formation of HMF was evaluated by means of a model solution. The results showed that a(w) and sugar concentration are both determinant in the formation of HMF in grape must. Sugar concentration influences the reaction by supplying substrates; low a(w) enhances the formation of HMF by changing the equilibrium in the dehydration step of the reaction.     

不同磁场环境和培养基对拟南芥种子诱导愈伤组织的影响

为探明磁场强度和培养基成份在拟南芥愈伤组织诱导中的作用,以野生型拟南芥(Columbia型)种子为材料,研究3种磁场环境(亚磁场、地磁场和中强磁场)、4种诱导培养基(NB1、NB2、MS1、MS2)条件下愈伤组织的诱导情况。结果显示,与地磁场相比,亚磁场对接种后第3～6天愈伤组织的发生有明显的抑制作用,中强磁场有明显的促进作用;在接种后第3～7天,NB2、MS2比NB1、MS1培养基促进了愈伤组织形成,且差异显著。愈伤组织净重反映了细胞增殖的状态,统计结果显示,与地磁场相比,亚磁场对细胞增殖没有明显影响,而中强磁场有显著的抑制作用;NB2、MS2比NB1、MS1培养基促进了细胞增殖,且差异显著。以上试验结果说明,磁场强度和培养基成份的变化对拟南芥种子诱导愈伤组织过程中第3～7天的愈伤组织形成和细胞增殖有不同程度影响,但2因素不存在交互作用,且2种因素对15d后的愈伤组织总出愈率无显著影响。6-BA对早期的愈伤组织诱导和细胞增殖均有促进作用,而且在地磁和亚磁场中,添加有6-BA的NB2和MS2培养基所诱导的愈伤组织色泽鲜亮,致密度适中,利于后续继代培养。     

The formation of wine lactone from grape-derived secondary metabolites

Wine lactone (i.e., 3a,4,5,7a-tetrahydro-3,6-dimethylbenzofuran-2(3H)-one, 1a/1b) was formed hydrolytically at wine pH from both racemic (E)-2,6-dimethyl-6-hydroxyocta-2,7-dienoic acid (3) and the corresponding glucose ester 2a at 45 °C but at room temperature was only formed from the acid 3. The glucose ester does not appear to be a significant precursor for the formation of wine lactone in wine. The slow formation of wine lactone from the free acid 3 indicates that the acid is not likely to be an important precursor to wine lactone in young wines unless present in high concentration (? 1 mg/L), but could be a significant precursor to wine lactone in wine that is several years old. The wine lactone formed in hydrolysates of the (6R)-enantiomer of 3 was partially enriched in the (3S,3aS,7aR)-enantiomer 1a when the hydrolysis was conducted at pH 3.2 and 100 °C in a closed vessel or under simultaneous distillation-extraction (SDE) conditions, and the enantiomeric excess (ee) varied from 5 to 22%. Hydrolysis of (6R)-3 in sealed ampules at 45 °C and at pH 3.0, 3.2, or 3.4 gave near-racemic wine lactone, but when the hydrolyses were conducted at room temperature, the product was enriched in the (3S,3aS,7aR)-enantiomer 1a and the ee was greater at higher pH (up to 60% at pH 3.4).     

Aroma biosynthesis in strawberry: s-adenosylmethionine:furaneol o-methyltransferase activity in ripening fruits

Among the most important volatile compounds in the aroma of strawberries are 2,5-dimethyl-4-hydroxy-3(2H)-furanone (Furaneol) and its methoxy derivative (methoxyfuraneol, mesifuran). Three strawberry varieties, Malach, Tamar, and Yael, were assessed for total volatiles, Furaneol, and methoxyfuraneol. The content of these compounds sharply increased during fruit ripening, with maximum values at the ripe stage. An enzymatic activity that transfers a methyl group from S-adenosylmethionine (SAM) to Furaneol sharply increases during ripening of strawberry fruits. The in vitro generated methoxyfuraneol was identified by radio-TLC and GC-MS. The partially purified enzyme had a native molecular mass of approximately 80 kDa, with optimum activity at pH 8.5 and 37 degrees C. A high apparent K(m) of 5 mM was calculated for Furaneol, whereas this enzyme preparation apparently accepted as substrates other o-dihydroxyphenol derivatives (such as catechol, caffeic acid, and protocatechuic aldehyde) with much higher affinities (K(m) approximately 105, 130, and 20 microM, respectively). A K(m) for SAM was found to be approximately 5 microM, regardless of the acceptor used. Substrates that contained a phenolic group with only one OH group, such as p-coumaric and trans-ferulic acid, as well as trans-anol and coniferyl alcohol, were apparently not accepted by this activity. It is suggested that Furaneol methylation is mediated by an O-methyltransferase activity and that this activity increases during fruit ripening.     

Improved synthesis and deployment of (2S,3R)-2-(2Z,5Z-octadienyl)-3-nonyloxirane, a pheromone of the pink moth, Lymantria mathura

We report two new syntheses of (2S,3R)-2-(2Z,5Z-octadienyl)-3-nonyloxirane, the main sex pheromone component of the pink moth, Lymantria mathura. The key step in the first route was the construction of (Z,Z)-1-bromo-1,4-heptadiene (6), which was coupled in the final step with 2-iodomethyl-3-nonyloxirane 4 via a Grignard reaction. The second approach employed alkylation of 1,4-heptadiynyllithium with epoxy triflates 7 in ether/hexane and provided the pheromone in >/=37% overall yield from alcohol 2. The 4:1 ratio of pheromone enantiomers, reportedly the most attractive to pink moth males, can be directly crafted from appropriately selected Sharpless asymmetric epoxidation conditions.     

5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)- 1,2-isoxazoline as a useful rice herbicide

5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)-1,2-isoxazoline derivative was synthesized, and its herbicidal activity was assessed under glasshouse and flooded paddy conditions. 5-(2,6-Difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)-1,2-isoxazoline demonstrated good rice selectivity and potent herbicidal activity against annual weeds at 125 g of a.i. ha(-1) under greenhouse conditions. Soil application of this compound showed complete control of barnyard-grass to the fourth leaf stage at 250 g of a.i. ha(-1). Field trials indicated that this compound controlled annual weeds rapidly with a good tolerance on transplanted rice seedlings by post-emergence and soil application. This compound showed a low mammalian and environmental toxicity in various toxicological tests.