Fast determination of Sudan dyes in chilli tomato sauces using partial filling micellar electrokinetic chromatography

A new method based on partial filling micellar electrokinetic chromatography (MEKC) for the quantitative determination of Sudan dyes (I, II, III, and IV) in chilli sauces is presented. The separation is achieved filling 25% of the capillary with a MEKC buffer composed of 40 mM NH(4)HCO(3), 25 mM sodium dodecyl sulfate, and 32.5% (v/v) acetonitrile (ACN). The rest of the capillary is filled using a capillary zone electrophoresis (CZE) buffer composed of 40 mM NH(4)HCO(3) and 32.5% (v/v) ACN. Under optimized conditions, the azo dyes are baseline separated in less than 8 min with limits of detection ranging from 0.57 to 0.71 μg mL(-1) (S/N > 3). Using an internal standard, the repeatability of the quantitative determination is improved almost four times. The applicability of the method for rapid screening and determination of Sudan dyes is corroborated by analyzing spiked chilli sauce samples with recoveries from 85 to 99%. The reported conditions are demonstrated to be compatible with mass spectrometry detection.     

Enantioseparation of imazalil residue in orange by capillary electrophoresis with 2-hydroxypropyl-beta-cyclodextrin as a chiral selector

Chiral resolution of imazalil, a fungicide, was performed by capillary electrophoresis (CE) using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector. Factors affecting the chiral resolution and migration time of imazalil were studied. The optimum running conditions were found to be 5 mM ammonium dihydrogenphosphate-50 mM phosphate buffer (pH 3.0) containing 4 mM 2-hydroxypropyl-beta-cyclodextrin with an effective voltage of +25 kV at 20 degrees C using direct detection at 200 nm. Under these conditions, the resolution (Rs) of racemic imazalil was approximately 6. The extraction of imazalil from orange samples was done with acetonitrile under basic conditions. The extract was purified with a solid-phase extraction cartridge (Sep-Pak plus PS-2) and was analyzed by the above CE method. Eight orange samples were analyzed, and imazalil was detected in seven samples. In four of these seven oranges, the level of (-)-imazalil was the same as that of (+)-imazalil, but in the other three oranges, the level of (-)-imazalil was found to be lower than that of (+)-imazalil, suggesting that (-)-imazalil was degraded more quickly than (+)-imazalil in oranges.     

Application of micellar electrokinetic capillary chromatography to the analysis of uncharged pesticides of environmental impact

A test mixture of five pesticides and metabolites (naphthalene acetamide, carbaryl, 1-naphthol, thiabendazole, and carbendazime) has been investigated by capillary electrophoresis with an ultraviolet diode array detector. These compounds were separated in <10 min by micellar electrokinetic capillary chromatography (MEKC). MEKC was performed in 30 mM ammonium chloride/ammonia buffer (pH 9.0) containing 15 mM sodium dodecyl sulfate. The lowest detection limit was obtained for the insecticide carbaryl (0.22 microg mL(-)(1)) and the highest for its metabolite 1-naphthol (1.13 microg mL(-)(1)). This method was applied to the analysis of the pesticides in cultivated vegetables such as cucumbers, which were extracted with a liquid-liquid extraction procedure, obtaining recovery percentages ranging from 90.1 to 110.2%.     

A capillary electrophoresis laser-induced fluorescence method for analysis of potato glycoalkaloids based on a solution-phase immunoassay. 1. Separation and quantification of immunoassay products

Solution-phase immunoassays are typically faster and more precise than ELISAs. This research developed a solution-phase for the immunoassay of potato glycoalkaloids (GAs) based on quantification by capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection. Solanidine coupled to 4'-(aminomethyl)fluorescein and a polyclonal antibody solution were used as the immunoreagents. Unbound fluorescent solanidine was detected by CE-LIF (excitation 488 nm, emission 520 nm). Optimum resolution of immunoassay products was achieved with a buffer consisting of 50 mM phosphate, 10% (v/v) methanol, and 1.5 mM SDS, pH 7.5. A plot of signal vs log [GA] produced a sigmoidal curve typical of immunoassays. Analysis of extracts of sprouted Yukon Gold potato tubers and nonsprouted Yukon Gold tubers resulted in total [GA] of 98 microg/g (RSD 9%) and 55 microg/g (RSD 9%), respectively. The findings indicated that CE-LIF coupled with a solution-phase immunoassay can be used to quantify total GA in potatoes.     

Epimeric separation of L-ascorbic acid and D-isoascorbic acid by capillary zone electrophoresis

Capillary zone electrophoresis (CZE) was used for separation of L-ascorbic acid (L-AA) and D-isoascorbic acid (D-IAA) in a model system. The effects of borate buffer concentration (0.05-0.25 M) and pH (pH 7.5-9.0) on migration time, resolution (Rs), and theoretical plates (N) were investigated. The migration times of L-AA and D-IAA increased with the increasing pH of carrier electrolyte (0.2 borate buffer), and the resolutions (Rs) of L-AA and D-IAA were calculated to be 12.98 at pH 9.0. Concentrations of borate buffer (pH 9.0) increased the Rs values of L-AA and D-IAA, and buffer concentrations >0.1 M were found to be effective for separation of L-AA and D-IAA. Methanol in the carrier electrolyte was also influential in improving the separation of L-AA and D-IAA, which increased with the increasing concentrations (0-10%) of methanol. The optimal separation conditions for L-AA and D-IAA were as follows: carrier electrolyte, 0.2 M borate buffer (pH 9.0); applied voltage, 25 kV, with an uncoated fused silica capillary, 75 microm (i.d.) x 57 cm.     

添加菌体蛋白和尿素对玉米秸秆厌氧发酵的影响

以玉米秸秆为原料的厌氧发酵,碳氮比（C/N）一直是影响其发酵的主要问题之一。该文针对玉米秸秆的C/N进行了研究,通过对比试验的方法,以尿素和菌体蛋白作为氮源添加剂配制成C/N分别为25/1和35/1的秸秆发酵料液,进行30d的厌氧发酵试验。结果表明,以尿素和菌体蛋白作为氮源调整C/N时,C/N为35/1时的产气量要好于C/N为25/1;与秸秆单独发酵相比,C/N为35/1的添加菌体蛋白试验组R2和添加尿素试验组R4的总产气量分别提高了31.43%和2.69%。此外,添加菌体蛋白和尿素能有效抑制秸秆体积的漂浮膨胀,对料液起到酸碱缓冲的作用。菌体蛋白作为秸秆发酵添加剂应用在沼气工程中可以变废为宝,提高经济效益。     

氮素形态对解磷细菌发酵液解磷效果及小白菜生长和品质的影响

采用盆栽试验,研究了施用硝态氮、铵态氮对解磷细菌发酵液不同组分解磷活性及其对小白菜产量、品质的影响。结果表明,施用相同氮肥时,浇灌解磷细菌原发酵液处理 T3的作用效果明显优于菌体悬浮液T2及菌液上清T1;以硝态氮为底肥时,施用发酵原液的土壤有效态磷含量较高,小白菜产量最高,品质明显优化,即硝态氮含量下降, Vc、可溶性糖、可溶性蛋白、纤维素以及植株磷含量显著增加,且增加变幅均高于以铵态氮为底肥的效果。由此可知,硝态氮与解磷细菌发酵原液配施时, W1菌株解磷效果更好,可能是解磷菌株W1最适的氮源。因此,种植小白菜时直接向其根际施用解磷细菌发酵液并配施硝态氮肥,更有利于提高小白菜的品质及产量。     

Capillary Electrophoresis as a Tool for Evaluating Lactic Acid Production from Sorghum

Sorghum is an underutilized resource for the production of bioindustrial chemicals like lactic acid. Capillary electrophoresis (CE) was tested to monitor the fermentation process, i.e., quantify the amount of lactic acid and by‐products in the fermentation broth using phosphate buffer pH 6.25 containing the electroosmotic flow modifier cetyltrimethylammonium bromide (CTAB). High levels of calcium carbonate were required during fermentation to stabilize the pH and these caused considerably prolonged migration times in CE. A 1:10 dilution of the samples with water was the best way to reduce salt load and thus conductivity in the sample plug, and thus to eliminate the problem of prolonged migration times. Further improvements were achieved by rinsing the capillary with HCl and water after each run, rather than NaOH and water. HCl might more efficiently remove Ca²⁺ ions from the capillary surface. The fermentation broth studied was based on liquefied sorghum inoculated with Rhizopus oryzae. The main product was lactic acid (24.60 ± 0.56 g/L) and a significant by‐product was fumaric acid (1.07 ± 0.04 g/L).     

Enzymatic Process for Corn Dry‐Grind High‐Solids Fermentation

A modified dry‐grind process that combined the use of conventional amylases (glucoamylase [GA]), phytase, and granular starch hydrolyzing enzymes (GSHE) to achieve low liquefaction viscosities and low glucose concentrations during simultaneous saccharification and fermentation (SSF) with a high slurry solids content (>33% w/w) was developed. Doses of GSHE and GA were optimized for the modified process. At 35% solids content, the modified process had 80% lower slurry viscosity, 24% lower peak glucose concentration, 7.5% higher final ethanol concentration, and 51% higher fermentation rate compared with the conventional dry‐grind process. At 40% solids content, the modified process had lower viscosities, lower peak and residual glucose concentrations, and higher ethanol concentrations than the conventional process; however, the results were in contrast to those for 35% solids content. At 40% solids content, SSF did not run to completion for conventional or modified processes, and more than 2.5% w/v of residual glucose was left in the fermentation broth. Final ethanol concentration achieved with the modified process at 40% solids content was 19.5% v/v, similar to the ethanol concentration achieved with the modified process at 35% solids content. At 35% slurry solids content, a GSHE level of 1.25 μL/g db of corn and a GA level of 0.25 μL/g db of corn were selected as optimum enzyme doses for the modified process.     

Accurate determination of oosporein in fungal culture broth by differential pulse polarography

A simple and accurate differential pulse polarographic method has been developed for the determination of oosporein in the culture broth of the fungus Beauveria brongniartii. This hydroxybenzoquinone derivative is the only major secondary metabolite secreted by this entomopathogenic fungus, which is used as biological pest control agent (BCA) against Melolontha melolontha larvae. It can be found in the host organism as well as in the formulated product. The polarographic behavior of oosporein was examined in various buffer systems over the pH range 3-10. In Britton-Robinson buffer/methanol solution (3:7 v/v, pH 5.5) the differential pulse polarograms exhibited reproducible peaks at E(p) = -0.18 V vs silver/silver chloride/potassium chloride (3 M). Under these conditions, a plot of peak height vs concentration of oosporein was found to be linear over the range 5.9 x 10(-)(7) to 2.5 x 10(-)(5) M (0.18-7.74 microg mL(-)(1); r = 0.9998). The detection limit was calculated to be 54 ng mL(-)(1). To evaluate the concentration of oosporein, the standard addition method was applied. The analysis of oosporein in the culture broth led to a mean value of 524.9 microg mL(-)(1) broth with a relative standard deviation (S(rel)) of +/-2.6%. The proposed polarographic method is accurate, not time-consuming, and it is of low cost because no separation steps are necessary.     

生物表面活性剂产生菌的分离培养及其产物特性研究

从加油站受石油污染的土壤中分离到1株高效产生物表面活性剂的菌株,经鉴定为气生单孢菌(Aeromonas sp.)。并对其发酵液表面活性剂性能及稳定性进行分析,最佳条件下发酵液的表面张力可由70.2 mN/m 降至28.9 mN/m,对温度、pH、矿化度有较好的稳定性,维持85 %以上油水乳化液稳定时间达到350 h。对其发酵条件进行了优化,确定表面活性剂产生最适C源为2 %甘油,以硝酸盐和铵盐作为N源对表面活性剂产生无影响。对其产生物表面活性剂进行提纯,经TLC分析初步确定该表面活性剂为糖脂类化合物。     

固体稀释倍数和超声波对豆渣厌氧发酵酸化过程的影响

为了优化超声波对固体废弃物酸化过程的调节作用,缓解酸化产物抑制,该文研究超声波处理时不同稀释倍数对基质性质的影响及对处理后基质发酵性能的影响,根据基质的挥发性有机酸去除率和发酵性能选取较佳的超声波处理参数。试验结果表明,超声波对不同浓度酸化基质的处理可去除有机酸中80%以上的非电离有机酸,减小对微生物的毒害,同时可减小基质的颗粒尺寸。处理后的基质经10 d发酵以评价不同浓度条件下的酸化性能,当基质挥发性有机酸质量浓度在18.8 g/L左右时,将基质稀释至固体质量浓度为46 g/L后再进行超声波处理,基质的挥发性有机酸增长率和挥发性固体降解率可分别从仅经过超声波辐射处理的51.8%、14.7%提高到197.4%、26.8%,表现出较佳的处理性能。可为超声波调节固体废弃物酸化过程提供参考。     

复合益生菌发酵液的功能特性及对对虾诱食效果

为探求复合菌发酵液的多种功能,该试验选择具有脱氮、产酶、抑菌等优良性能的酵母菌、乳酸菌及芽孢杆菌各1株,利用已优化的HJ培养基进行共培养,实时监测发酵过程,分时段取样,对复合菌发酵液的脱氮、产酶、抑菌、培藻及诱食等功能进行研究。结果表明,酿酒酵母菌NJ-02、屎肠球菌SC-01及枯草芽孢杆菌M7-1能够在HJ培养基中进行共发酵,连续培养24 h后3株微生物活菌数量分别达到3.88×10~8、2.41×10~(10)、5.38×10~9 CFU/mL。复合菌发酵液的脱氮、培藻性能同复合菌中枯草芽孢杆菌M7-1的活菌数相关较大,发酵至16h其降解亚硝态氮和培藻性能最为理想,亚硝态氮降解率为89%,并使小球藻叶绿素a质量分数提升49.6%。复合菌发酵液具有同枯草芽孢杆菌M7-1相当的产酶(蛋白酶、淀粉酶、纤维素酶)活性及同屎肠球菌SC-01相当的抑菌(副溶血弧菌)活性。复合菌发酵液饲喂对虾,诱食效果明显好于对照组(P0.05),同化学诱食剂二甲基-β-丙酸噻亭(dimethyl-beta-propiothetin hydrochloride, DMPT)差异不显著,其肠道中乳酸菌、酵母菌数目显著高于对照组及化学诱食剂组(P 0.05)。该研究所制备的脱氮、培藻、抑菌及诱食功能复合菌发酵液,为可持续生态水产养殖提供了新的微生物资源和技术方法。     

Liquid chromatographic determination of ochratoxin A in coffee beans and coffee products

A liquid chromatographic method for the determination of ochratoxin A in coffee beans (green and roast), instant coffee, and coffee drink is described. The sample is subjected to extraction with methanol-1% aqueous sodium bicarbonate (1 + 1) and C18 cartridge cleanup. The extract is chromatographed on a Nucleosil 5C18 column with a mobile solvent of acetonitrile-water-0.2M phosphate buffer pH 7.5 (50 + 47 + 3) containing 3 mM cetyltrimethylammonium bromide as an ion-pair reagent. Ochratoxin A is detected with a fluorometer (excitation 365 nm, emission 450 nm). The sensitivity was increased 20-fold by using ion-pair resolution. The detection limits corresponded to 2 micrograms/kg for coffee beans, 5 micrograms/kg for instant coffee, and 0.2 microgram/kg for coffee drink. The recoveries from coffee products were generally better than 80.7% and the relative standard deviations were 3.43-5.93%. The peak coinciding with ochratoxin A can be confirmed by treatment using alcohol (methanol, ethanol, or n-propanol) and H2SO4.     

Separation and Determination of Three Phenolic Xenoestrogens in Industrial Wastewater by Micellar Electrokinetic Chromatography on Polydimethylsiloxane Microchip

The separation on microchip provides the advantages including high efficiency, increased throughput, reduced quantities of hazardous materials, cost saving, relatively facile instrumentation, improved portability, etc. A technique of micellar electrokinetic chromatography (MEKC) coupled with amperometric detection has been actualized on a polydimethylsiloxane microchip for the rapid separation and determination of three phenolic xenoestrogens as octylphenol (OP), 4-nonylphenol (4-NP), and bisphenol A (BPA). The baseline separation of these phenolic xenoestrogens is successfully obtained within 55?s under the optimized MEKC conditions with borate running buffer of pH?8.0 containing sodium dodecyl sulfate and ??-cyclodextrin. The linear range for OP, 4-NP, and BPA are 20?C1,000, 15?C1,000, and 20?C1,000???g/L with the detection limit of 5.0, 4.0, and 3.0???g/L, respectively. The present method is successfully applied for the determination of these phenolic xenoestrogens in some industrial wastewater samples from mainland of China with the recoveries ranged from 90.2 to 109.4?%.     

Sorghum Protein Extraction by Sonication and Its Relationship to Ethanol Fermentation

The objectives of this research were to develop a rapid method for extracting proteins from mashed and nonmashed sorghum meal using sonication (ultrasound), and to determine the relationships between the levels of extractable proteins and ethanol fermentation properties. Nine grain sorghum hybrids with a broad range of ethanol fermentation efficiencies were used. Proteins were extracted in an alkaline borate buffer using sonication and characterized and quantified by size‐exclusion HPLC. A 30‐sec sonication treatment extracted a lower level of proteins from nonmashed sorghum meal than extracting the proteins for 24 hr with buffer only (no sonication). However, more protein was extracted by sonication from the mashed samples than from the buffer‐only 24‐hr extraction. In addition, sonication extracted more polymeric proteins from both the mashed and nonmashed samples compared with the buffer‐only extraction method. Confocal laser‐scanning microscopy images showed that the web‐like protein microstructures were disrupted during sonication. The results showed that there were strong relationships between extractable proteins and fermentation parameters. Ethanol yield increased and conversion efficiency improved significantly as the amount of extractable proteins from sonication of mashed samples increased. The absolute amount of polymeric proteins extracted through sonication were also highly related to ethanol fermentation. Thus, the SE‐HPLC area of proteins extracted from mashed sorghum using sonication could be used as an indicator for predicting fermentation quality of sorghum.     

拮抗菌DG1的鉴定及其对大白菜菌核病菌的抑制作用研究

为减少化学农药的使用,丰富生物防治菌株资源库,在分离大白菜菌核病菌的过程中,筛选到一株对其具有抑制作用的拮抗菌,命名为DG1.本研究通过形态观察、生理生化特征检测及16S rDNA序列分析,对拮抗菌DG1进行种属鉴定;通过测量菌落直径,研究DG1发酵液及挥发性物质对大白菜菌核病菌的抑制作用;通过平板对峙法测定拮抗菌DG...     

Combined use of supercritical fluid extraction, micellar electrokinetic chromatography, and reverse phase high performance liquid chromatography for the analysis of antioxidants from rosemary (Rosmarinus officinalis L.)

Antioxidants from rosemary were determined by the combined use of supercritical fluid extraction (SFE) prior to reverse-phase high-performance liquid chromatography (RP-HPLC) or micellar electrokinetic chromatography (MEKC). The separation of antioxidants found in the SFE fractions was achieved by using a new MEKC method and a published HPLC procedure, both with diode array detection. The characterization of the different antioxidants was further done by HPLC-mass spectrometry. Advantages and drawbacks of HPLC and MEKC for analyzing the antioxidants found in the different extracts are discussed. From the results it is concluded that HPLC renders higher peak area and is better in its reproducibility than MEKC; both techniques provide similar analysis time reproducibility. The main advantage of MEKC is its much higher separation speed, which is demonstrated to be useful for the quick adjustment of SFE conditions, allowing rosemary fractions of higher antioxidative power to be obtained quickly. Moreover, the possibilities of this approach for following the degradation of antioxidants are discussed.     

Enantioseparation of isoxanthohumol in beer by hydroxypropyl-gamma-cyclodextrin-modified micellar electrokinetic chromatography

Chiral resolution of isoxanthohumol (IX) in beer samples was performed by hydroxypropyl-gamma-cyclodextrin-modified micellar electrokinetic chromatography. The optimum running conditions were found to be 20 mM phosphate buffer (pH 7.0) containing 45 mM hydroxypropyl-gamma-cyclodextrin and 100 mM sodium dodecyl sulfate with an effective voltage of +20 kV at 20 degrees C using direct detection at 210, 295, and 370 nm. IX was detected in 12 beer samples and the total levels of (+)- and (-)-IX ranged from 0.15 to 1.4 mg/L. But the amount of xanthohumol (XN) was below the detection limit (0.017 mg/L). Each level of (-)-IX was almost the same as that of (+)-IX, suggesting that IX was a racemic mixture in these beer samples. The racemization of IX in beer could be attributed to the production of a racemic mixture from XN during boiling and to the fact that IX enantiomers were easily interconverted.