Enzymatic preparation of kaempferol from green tea seed and its antioxidant activity

Among the flavonols in green tea, kaempferol has many biological activities but kaempferol of plant origin is too expensive to be used in commercial products. Recently, we confirmed that green tea seed (GTS) contained a reasonable amount of kaempferol glycoside. After conducting structure analysis, two kaempferol glycosides were identified, kaempferol-3-O-[2-O-beta-D-galactopyranosyl-6-O-alpha-L-rhamnopyranosyl]-beta-D-glucopyranoside (compound 1) and kaempferol-3-O-[2-O-beta-D-xylopyranosyl-6-O-alpha-L-rhamnopyranosyl]-beta-D-glucopyranoside (compound 2), respectively. Also, a commercially useful method for kaempferol preparation was suggested by enzymatic hydrolysis using these two flavonoids. After several enzyme reactions were performed for the complete bioconversion of compounds 1 and 2 to kaempferol, we found that the optimum enzyme combination was reaction with beta-galactosidase and hesperidinase. Finally, we produced pure kaempferol with over 95% purity. We also compared the antioxidant effect of these two GTS flavonoids and its aglycone, kaempferol. Kaempferol is a more efficient scavenger of 1,1-diphenyl-2-picrylhydrazyl radicals and a better inhibitor of xanthine/xanthine oxidase than the two glycosides.     

Oxidative degradation of triazine derivatives in aqueous medium: a radiation and photochemical study

Pulse and steady state radiolysis techniques have been used to determine the bimolecular rate constants and to investigate the spectral nature of the intermediates and the degradation induced by hydroxyl radicals ((*)OH) with 1,3,5-triazine (T), 2,4, 6-trimethoxy-1,3,5-triazine (TMT), and 2,4-dioxohexahydro-1,3, 5-triazine (DHT) in aqueous medium. A competitive kinetic method with KSCN as the (*)OH scavenger was used to determine the rate constants for the reaction of (*)OH with T, TMT, and DHT. The bimolecular rate constants are 3.4 x 10(9), 2.06 x 10(8), and 1.61 x 10(9) dm(3) mol(-)(1) s(-)(1) respectively, for T, TMT, and DHT at pH approximately 6. The transient absorption spectra obtained from the reaction of (*)OH with T, TMT, and DHT have single absorption maxima at 320, 300, and 300 nm, respectively, and were found to undergo a second-order decay. The formation of TOH(*) [C(6)OH-N(5)-yl radical], TMTOH(*) [N(5)OH-C(6)-yl radical], and DHT(*) [C(6)-yl radical] is proposed from the initial attack of (*)OH with T, TMT, and DHT, respectively. A complete degradation of TMT (10(-3) mol dm(-3)) was obtained after absorbed doses of 5 kGy in N(2)O-saturated solutions and 16 kGy in aerated solutions. A similar degradation pattern was obtained with DHT in N(2)O-saturated solutions. Complete degradation was observed with an absorbed dose of 7 kGy. On the basis of the results from both pulse and steady state radiolysis, a possible reaction mechanism involving (*)OH-mediated oxidative degradation is proposed. A complete photodecomposition of DHT was also observed in the presence of ferric perchlorate using ultraviolet light at low pH. Photoinduced electron transfer between Fe(III) and DHT in the Fe(III)-DHT complex and subsequent formation of DHT(*) are proposed to be the major processes that lead to the complete degradation of DHT at pH 3.     

Radiolysis of quercetin in methanol solution: observation of depside formation

Radiolysis of the flavonol quercetin, a natural antioxidant, was performed in methanol. The degradation process was followed by HPLC analyses. The major product was identified as a depside (Q1) by NMR and LC-MS. The G(Q1) radiolytic factor was plotted versus the initial concentration of quercetin. This radiolytic process was attributed to the CH3O* radicals presented in the irradiated medium. The proposed mechanism invoked a stereospecific oxidation of the 3-hydroxyl group of quercetin which led to C-ring opening and to the formation of the depside Q1. In presence of water, Q1 was transformed into another depside, Q2, by an inverse esterification reaction. A chemical equilibrium was observed between Q1 and Q2. The comprehension of the radiolytic process of quercetin in methanol solution is of importance. Indeed, the same type of oxidative reactions could occur on flavonoids during preservation of food by ionizing radiation.     

辐照对黄原胶分子量及抗氧化活性的影响

以0~500kGy 60Co γ射线对固态黄原胶进行辐照,研究辐照对黄原胶分子量及还原力、清除1,1-二苯基-2-苦基肼自由基（DPPH·）、超氧自由基（O2-·）及羟自由基（·OH）能力等抗氧化活性的影响。研究表明,10kGy的辐照可使黄原胶粘均分子量从1.71×106增加到2.23×106,超过10kGy后,黄原胶粘均分子量随辐照剂量的增大逐渐减少,500kGy时降为1.59×104。低于10kGy辐照,黄原胶的还原力、对DPPH·和对O2-·的清除率与剂量呈负相关,与对照相比,还原力降低了3.16%,对DPPH·和O2-·的清除率分别从17.37%和18.18%减少到16.87%和8.51%,增大辐照剂量,黄原胶的还原力和对DPPH·和O2-·的清除率与剂量呈正相关,500kGy时黄原胶的还原力比对照增加164.22%,对DPPH·和O2-·的清除率升高至60.79%和30.06%;对·OH清除能力与辐照剂量呈负相关,清除率从0kGy时的9.29%降低到500kGy时的3.63%。辐照可以引起黄原胶聚合或降解,从而导致其分子量及抗氧化活性发生显著变化。     

γ射线辐照降解木聚糖的机理研究

为了探讨γ射线辐照降解半纤维素的机理,以木聚糖为模型化合物,经0、200、400、600、800、1 000、1 200 k Gy剂量的辐照后,采用扫描电镜(SEM)、傅里叶拉曼光谱(FT-Raman)、气相色谱质谱联用仪(GC-MS)研究其表观形貌结构、分子结构及降解产物的变化特征。结果表明,γ射线辐照处理后木聚糖表观形貌结构发生破坏,多孔性增加;随着辐照剂量升高,木聚糖3 354 cm~(-1)处拉曼光谱峰强度降低且峰位发生红移,1 122 cm~(-1)和1 093 cm~(-1)处拉曼光谱峰强度逐渐减弱,表明木聚糖分子中的氢键、糖苷键发生破坏;木聚糖降解产物的GC-MS分析表明,辐照处理后木聚糖乙酸乙酯提取液中共检测到18种脂肪酸类化合物、5种糖类化合物和2种呋喃类化合物,其相对含量分别在1 200、800和1 200 k Gy达到最大值(16.59%、1.46%和1.03%)。本研究结果为木聚糖γ射线辐照降解产物全利用提供了支持。     

Antioxidative effect of kaempferol and its equimolar mixture with phenyltin compounds on UV-irradiated liposome membranes

The work investigates the possibility of the protective action of kaempferol on phosphatidylcholine liposome membranes exposed to the pro-oxidative action of diphenyltin dichloride (DPhT) and triphenyltin chloride (TPhT) induced by UV radiation (lambda = 253.7 nm). The concentrations of kaempferol and its equimolar mixtures with DPhT and TPhT were determined so that they induce 50% inhibition in oxidation of liposomes irradiated with UV. They are 11.6, 10.0, and 4.5 microM/L, which constitute the following sequence of antioxidative activity: kaempferol/triphenyltin > kaempferol/diphenyltin > kaempferol. This relationship is confirmed by the results on the antiradical ability of kaempferol and its mixtures with DPhT and TPhT toward the free radical 2,2-diphenyl-1-picrylhydrazyl. Similar sequences obtained in both studies suggest a possible mechanism of the antiradical action of the mixtures as free radical scavengers. Kaempferol's ability, then documented, to form complexes with phenyltins indicates (a) a possible way to liquidate the peroxidation caused by the free radical forms of phenyltins and (b) the stabilizing role of chelating in the antioxidative action of the kaempferol/phenyltins. The differentiation in the action of the compounds studied may, among others, result from different localizations in the liposome membrane, which is indicated by the results of the fluorometric studies.     

水溶液和芦笋中毒死蜱的辐照降解

为寻求一种能有效降解毒死蜱残留的技术方法,本文采用60Co γ射线辐照水溶液和芦笋中的毒死蜱,并对比了毒死蜱的辐照降解率。结果表明,辐照剂量在0～12kGy时,水溶液中毒死蜱的降解率随剂量的增加而增大,4kGy的辐照可使其降解率达95.5%;芦笋中毒死蜱的辐照降解率偏低,8kGy辐照时降解率最高,达到30.0%。进一步研究表明,芦笋成分Vc、芦丁和总糖等的存在对辐照降解毒死蜱有抑制效应。     

Light-induced oxidative changes in a model dairy spread. Wavelength dependence of quantum yields

Light-induced formation of lipid peroxides in a water-in-oil emulsion based on purified rape-seed oil (80%) was found to increase with decreasing wavelength and to have the (apparent) quantum yields (1.1 +/- 0.1) x 10(-)(3) for 436 nm, (2.6 +/- 0.1) x 10(-)(3) for 405 nm, and (4.5 +/- 0.4) x 10(-)(3) for 366 nm irradiation, as determined after 12 h of exposure to monochromatic light of an approximate intensity of 10(18) quanta.min(-)(1).mL(-)(1) and related to total light absorption. Riboflavin (0.8 ppm) had no effect on lipid peroxidation, but photodegraded with a quantum yield ((1.5 +/- 0.3) x 10(-)(5) for 436 nm, (1.7 +/- 0.2) x 10(-)(5) for 405 nm and (1.39 +/- 0.09) x 10(-)(5) for 366 nm irradiation) independent of irradiation wavelength. beta-Carotene was only photodegraded to a minor extent, but protected riboflavin against photodegradation and the lipids against peroxidation for 436 and 405 nm irradiation (reduction in quantum yield three times for 4.5 ppm beta-carotene for lipid oxidation and more for riboflavin degradation), but not for 366 nm irradiation, where beta-carotene has an absorption minimum.     

Fractionation of lentil seeds (Lens culinaris Medik.) for insecticidal and flavonol tetraglycoside components

Crude methanol extracts from four cultivated varieties of mature lentil seeds (Lens culinaris Medik.) were found to possess antifeedant and insecticidal properties in laboratory tests with the rice weevil (Sitophilus oryzae L.), an insect pest of stored products. Flash chromatography with silica gel on active Diaion HP-20 methanol extracts gave flavonol, lysolecithin, soyasaponin, and peptide fractions, as determined by HPLC and electrospray ionization LC/MS. The flavonol fraction was shown by high-resolution NMR experiments to contain a mixture of kaempferol 3-O-beta-glucopyranosyl(1-->2)-O-[alpha-rhamnopyranosyl(1-->6)]-beta-galactopyranoside-7-O-alpha-rhamnopyranoside and, tentatively, kaempferol 3-O-beta-glucopyranosyl(1-->2)-O-[alpha-rhamnopyranosyl(1-->6)]-beta-glucopyranoside-7-O-alpha-rhamnopyranoside. These inactive tetraglycosides, although inseparable under the reported HPLC conditions, were detected by NMR spectroscopy in nearly equal proportions. Three lysolecithins were identical to those previously identified in pea extracts. Soyasaponin I (soyasaponin Bb) and soyasaponin VI (soyasaponin betag) were found in Diaion HP-20 methanol extracts. An insecticidal lentil peptide with a mass of 3881 Da, isolated from an Eston variety in small quantities by anion exchange chromatography, was related to the cysteine-rich pea albumin 1b class of botanical insecticides. Binary mixtures of the insecticidal lentil peptide and soybean soyasaponin I were synergistic in tests with S. oryzae.     

辐照与化学相结合协同降解壳聚糖研究

研究了单一辐照和协同处理对壳聚糖降解的影响。在协同降解中研究了辐照剂量、H2O2浓度、溶剂/溶质体积质量比、乙酸4个因素的作用。对前3个因素进行了方差分析和不同水平的多重比较,同时对降解产物的结构进行了红外光谱分析。结果表明协同降解效果较好,即15%H2O2、1.5%HAc、H2O2溶液/壳聚糖体积质量比4∶1、辐照剂量为110kGy可以使壳聚糖的分子量从8.7×105降低到104以下,且结构没有明显变化。     

辐照对发酵香肠品质特性的影响

本文以0、2、4、6kGy剂量60Co γ射线辐照发酵香肠,以水分活度、水分含量、pH值、含盐量、氨基酸总量、非蛋白氮、总氮、硫代巴比妥酸值、挥发性风味物质、肌浆蛋白与肌原纤维蛋白降解情况为指标,研究辐照对香肠品质的影响。结果表明：发酵香肠经不同剂量辐照后水分含量差异显著(P<0.05),氯化钠含量差异并不显著,而0、2kGy辐照后发酵香肠的pH值显著高于4、6kGy辐照,4kGy剂量辐照的发酵香肠中氨基酸总量显著高于2、6kGy处理的样品(P<0.05),4kGy剂量辐照香肠烯烃、酸和酮类物质含量最多,6kGy辐照组中醇类物质含量最多。辐照处理的发酵香肠中肌浆蛋白和肌原蛋白的降解情况低于对照。     

Evaluation of 2-alkylcyclobutanones in irradiated cured pork products during vacuum-packed storage

The 2-Alkylcyclobutanones (2-ACBs) content was determined in three Italian cured pork products (salame Milano, coppa, and pancetta) irradiated at different targeted irradiation doses (2, 5, and 8 kGy) during vacuum-packed storage. Among 2-ACBs, three different compounds were investigated, namely, 2-dodecylcyclobutanone, 2-tetradecylcyclobutanone, and 2-(tetradec-5'-enyl)cyclobutanone. 2-ACBs were absent from the nonirradiated samples, whereas their content increased with irradiation dose. Their presence was recorded occasionally at 2 kGy and constantly at higher irradiation doses (5 and 8 kGy). The plot of 2-ACBs content against targeted irradiation doses showed an exponential relationship. The effect of vacuum-packed storage time on the 2-ACBs content was dependent on the irradiation dose. During vacuum-packed storage for up to 60 days, the 2-ACBs content remained unchanged in the cured pork products irradiated at 2 and 5 kGy, whereas a significant increase was observed in the pork products irradiated at 8 kGy.     

建立谷物制品辐照杀虫工艺规范的研究

通过对国内外大量文献和补充试验数据的分析 ,确定了谷物制品辐照杀虫加工工艺规范的工艺参数和技术要求 :辐照前要求产品含水量小于 1 2 % ,无蛹和成虫期害虫 ,产品包装后立即辐照 ,辐照杀虫最低有效剂量为 0 3kGy ,各种谷物制品的最高耐受剂量在 0 5～ 0 8kGy之间。     

Mechanochemical-assisted extraction and antioxidant activities of kaempferol glycosides from Camellia oleifera Abel. meal

A mechanochemical-assisted extraction (MCAE) method was proposed and investigated for the fast extraction of two kaempferol glycosides (kaempferol-3-O-[2-O-β-D-galactopyranosyl-6-O-α-L-rhamnopyranosyl]-β-D-glucopyranoside and kaempferol-3-O-[2-O-β-D-xylopyranosyl-6-O-α-L-rhamnopyranosyl]-β-D-glucopyranoside) from Camellia oleifera Abel. meal. The effects of operating parameters in terms of NaOH content, grinding time, extraction time, and ratio of solution to solid were evaluated by means of response surface methodology (RSM). Under the optimal conditions with a ratio of material to NaOH of 20:1 (g/g), a milling time of 15 min, and a ratio of solution to solid of 20:1 (mL/g) for 60 min, the maximum extraction yields of the two kaempferol glycosides reached 13.34 and 13.83%, respectively. The antioxidant activity of kaempferol glycosides extract was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay and ferric thiocyanate (FTC) assay. Compared with the heat reflux extraction (HRE) method, the yield and the antioxidant activities of the extracts from MCAE with water as solvent were higher and stronger.     

60Co-γ射线和电子束辐照对鲜香菇保鲜效果的初步研究

为研究⁶⁰Co-γ和电子束2种射线的不同剂量对新鲜香菇采后保鲜效果的影响,本试验通过测定鲜香菇的硬度、呼吸强度、电导率、感官品质等指标,筛选2种辐照射线的最优剂量,并对比分析最优辐照处理下香菇细胞的微观结构。结果表明,⁶⁰Co-γ射线中,2 kGy剂量组香菇硬度降幅为37%,呼吸强度为10.81 mg·kg^-1·h^-1,相对电导率为0.268%,均小于其他剂量组;电子束射线中,3 kGy剂量组香菇硬度降幅为38%,呼吸强度为10.39 mg·kg^-1·h^-1,相对电导率为0.241%,均小于其他剂量组。综合感官分析可知, 3 kGy电子束组和2 kGy ⁶⁰Co-γ射线组香菇的保鲜效果分别优于同类射线其他剂量组。显微分析可知,与2 kGy ⁶⁰Co-γ射线相比,3 kGy电子束辐照能够较好地保持新鲜香菇的菌丝形态,延缓其进一步降解,从而延长其贮藏期。本研究探索了辐照在香菇保鲜过程中的可行性,为推动电子束辐照技术和⁶⁰Co-γ辐照技术在香菇保鲜领域的应用提供了可靠的理论依据与技术支持。     

Raman spectral analysis in the C-H stretching region of proteins and amino acids for investigation of hydrophobic interactions.

Raman spectra of amino acids showed complexity in the C-H stretching region (2800-3100 cm(-)(1)) attributed to diversity of CH, CH(2), and CH(3) groups in the side chains, ionization state, and microenvironment. The involvement of specific amino acids in the C-H stretching region of selected proteins, namely, lysozyme, alpha-lactalbumin, beta-lactoglobulin, and their binary mixtures, was investigated by deconvolution using maximum likelihood techniques. The main protein band near 2940 cm(-)(1) was attributed not only to aromatic and aliphatic amino acids but also to many other amino acids. A band near 3065 cm(-)(1) was assigned to aromatic residues, whereas bands near 2880 and 2900 cm(-)(1) corresponded primarily to aliphatic amino acids. Heating at 90 degrees C increased relative intensity near 2940 cm(-)(1) and decreased relative intensity at 2895-2902 cm(-)(1) for lysozyme and its mixtures with alpha-lactalbumin or beta-lactoglobulin. Additional bands at 2812 or 2838 and 3003 cm(-)(1) were observed after heating or in 8 M deuterated urea, reflecting changes upon denaturation.     

Effect of gamma-irradiation on degradation of alginate

The aqueous solution of alginate was irradiated by 60Co gamma-rays in the dose range of 10-500 kGy. To assess the effect of irradiation on the degradation of alginate, the irradiation-induced changes in the viscosity, molecular weight, color, monomer composition, and sequence were measured. The molecular weight of raw alginate was reduced from 300000 to 25000 when irradiated at 100 kGy. The degradation rate decreased and the chain breaks per molecule increased with increasing irradiation dose. The viscosity of irradiated alginate solution reached a near minimum as low as at 10 kGy. No appreciable color changes were observed in the samples irradiated at up to 100 kGy, but intense browning occurred beyond 200 kGy. The 13C NMR spectra showed that homopolymeric blocks, MM and GG, increased and the M/G ratio decreased with irradiation. Considering both the level of degradation and the color change of alginate, the optimum irradiation dose was found to be 100 kGy.     

Characterization of residual lignin after SO(2)-catalyzed steam explosion and enzymatic hydrolysis of Eucalyptus viminalis wood chips

The lignin component found in both water insoluble (WI) and water and alkali insoluble (WIA) fractions derived from SO(2)-impregnated steam-exploded eucalyptus chips (SEE) was isolated and characterized. Dioxane lignins with a sugar content lower than 2% (w/w) were obtained after each material was treated with commercial cellulases. The C9 formulas of both SEE-WI and SEE-WIA dioxane lignins were C(9)H(6.83)N(0.04)O(2.24)(OCH(3))(1.21)(OH(aro))(0.56)(OH(ali))(0. 77) and C(9)H(8.65)N(0.29)O(1.97)(OCH(3))(0.90)(OH(aro))(0. 46)(OH(ali))(1.02), respectively. The weight-average molecular weight (M(w)) of the SEE-WI lignin corresponded to 3.85 kDa, whereas the SEE-WIA lignin had an M(w) of 3.66 kDa for the same polydispersity of 2.4. The SEE-WIA lignin was shown to be more thermally stable than the SEE-WI lignin, requiring temperatures in the range of 520 degrees C for complete degradation. FTIR and (1)H NMR analyses of both untreated and peracetylated lignin fractions showed that (a) the alkali insoluble lignin contained a relatively higher degree of substitution in aromatic rings per C9 unit and that (b) alkaline extraction removed lignin fragments containing appreciable amounts of phenolic hydroxyl groups.     

Impacts of ionizing radiation on volatile production by ripening gala apple fruit

Apple (Malus x domestica Borkh., cv. Gala) fruit treated with 0.5 microL x L(-1) 1-methylcyclopropene (MCP) or air (non-MCP) for 12 h at 20 degrees C were exposed to gamma radiation at doses of 0, 0.44, 0.88, or 1.32 kGy at 23 degrees C and then stored at 20 degrees C. Production of volatile compounds was measured on the day of irradiation and 1, 3, 7, 14, and 21 days after irradiation. Both MCP treatment and irradiation inhibited ethylene production. MCP treatment reduced production of all volatile esters and alcohols detected, whereas irradiation inhibited production of most, but not all, esters and some alcohols by non-MCP-treated fruit. The inhibition of volatile production following irradiation increased with dose. Production of methyl and propyl esters was inhibited more than that of other esters following irradiation or MCP treatment. The impact of irradiation on production of esters and alcohols by MCP-treated fruit was minimal. Non-MCP-treated fruit irradiated at 0.44 kGy produced the most esters during the 21-day period at 20 degrees C following irradiation, and the ester production rate in these fruit was comparable to that of the nonirradiated fruit 21 days after irradiation. Fruit treated with doses higher than 0.44 kGy did not recover their ability to produce volatile compounds. These results indicate both MCP and ionizing radiation inhibit production of many aroma compounds produced by ripening apple fruit.