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1.
A standard method for the detection of Enterobacteriaceae was modified for the presumptive detection of Enterobacter sakazakii, and the modified method was validated in an interlaboratory trial with 16 laboratories from 8 European countries. The modification included a differential-elective medium for the isolation of E. sakazakii, consisting of nutrient agar (NA) supplemented with 4-methyl-umbelliferyl alpha-D-glucoside (alpha-MUG). A 25 g sample was added to 225 mL buffered peptone water. After incubation at 35 degrees or 37 degrees C for 16 or 20 h, 10 mL nonselective enrichment was transferred into 90 mL selective enrichment. The selective enrichment was streaked on violet-red bile glucose agar (VRBGA) and incubated at 37 degrees C for 24 h. It was streaked in parallel on NA plates supplemented with alpha-MUG at 50 mg/L and incubated at 25 degrees C for 16 h, and afterwards for an additional 24 h at room temperature in the dark. E. sakazakii appeared as vivid yellow colonies under normal light and showed blue/violet fluorescence under UV light on NA + alpha-MUG plates. Validation samples represented powdered infant formula without E. sakazakii (blanks) and with low (1-10 colony-forming units [CFU]/25 g) and medium (1-10 CFU/g) contamination levels. All samples contained Pseudomonas aeruginosa and Lactobacillus spp. as background flora. The specificity for blank samples was 100%. The sensitivity of the low contamination level was similar for VRBGA and NA + alpha-MUG, i.e., 66.7% (66.7% accordance, 53.9% concordance). For the medium level the sensitivities were 96.7% (93.3% accordance, 93.5% concordance) for VRBGA and 98.3% (96.9% accordance, 96.9% concordance) for NA + alpha-MUG.  相似文献   

2.
Eicosapentaenoic acid (EPA) Production byMortierella alpina ATCC 32222   总被引:2,自引:0,他引:2  
Mortierella alpina ATCC 32222 grew well at 11 degrees C, as well as at 25 degrees C in a liquid medium containing glucose or linseed oil and yeast extract. High Eicosapentaenoic acid (EPA) yield was obtained at 11 degrees C. M. alpina cells did not produce EPA at 25 degrees C in the absence of linseed oil, whereas at 11 degrees C, EPA accumulation was noted in the absence of linseed oil. When grown at 11 degrees C for 10 d in a medium containing 2% linseed oil as carbon source, the mycelium yielded 435 mg/L EPA (20 mg EPA/g dry mycelia) with 5.1% in lipid fraction. By gradually increasing the concentration of linseed oil to 4%, yield of biomass and EPA were increased to 43 g/L and 596 mg/L, respectively.  相似文献   

3.
The kinetics of thermal decomposition of 4-carboxyl-2,6-dinitrobenzenediazonium ion (CDNBD), an arenediazonium ion newly developed as a derivatizing reagent for drug analysis, are described. The arenediazonium ion, in an optimized concentrated sulfuric acid/orthophosphoric acid medium, was incubated for various time intervals at 30 degrees, 45 degrees, 55 degrees , 65 degrees , 75 degrees, and 85 degrees C. The amount of ion left after each time interval was quantified selectively by colorimetric assay at 490 nm, using mefenamic acid as a model diazo-coupling component. The rate constants for the decomposition were determined graphically. An Arrhenius plot was used to delineate the dependence of the rate constant on temperature and to predict the half-life at 25 degrees C and lower temperatures. The diazonium ion decomposed by first-order kinetics. The rate constants of decomposition, which increased progressively with temperature, were 3.18 +/- 0.41 x 10(-5), 1.19 +/- 0.07 x 10(-4), 4.87 +/- 0.15 x 10(-4), 12.88 +/- 0.73 x 10(-4), and 21.32 +/- 2.74 x 10(-4) (s(-1)) with corresponding half-lives of 363, 97.06, 23.72, 8.97, and 5.42 min at 30 degrees, 45 degrees, 55 degrees, 65 degrees, and 75 degrees C, respectively. CDNBD is highly stable in concentrated acid medium, with half-life values of about 10 h, 10 days, and 7.3 months at 25 degrees, 0 degrees, and -20 degrees C, respectively. The reagent stability profile shows that it could be readily adapted for routine applications in instrumental chemical analysis.  相似文献   

4.
A highly selective immunoaffinity column was obtained by coupling anti-papaverine polyclonal antibodies to CNBr-activated Sepharose 4B. It was found that the coupling efficiency and performance of the immunoaffinity column were greatly improved by prolonging the coupling reaction time from 3 h at 20 degrees C with shaking to incubation overnight at 4 degrees C after the 3 h shaking reaction. The pH and ionic strength were observed to be the most important factors that influence the binding of papaverine to the immunoaffinity column. Using 0.01 mol/L phosphate buffered saline (PBS, pH 8.3) and methanol-water (80:20, v/v) as the loading and eluting solutions, respectively, papaverine was first retained on the column and then quantitatively eluted out with a mean recovery of 86% at a loading concentration of 1 microg/mL. When applied to real samples of pericarpium papaveris and food products, the established immunoaffinity column showed high efficiency in removing the matrix interferences in the samples and satisfactory recovery results were obtained. The method was useful for extraction and purification of papaverine from related samples.  相似文献   

5.
Ehrlich ascites tumor cells spontaneously release cell surface material (glycocalyx) into isotonic saline medium. Exposure of these cells to tritium-labeled 4,4'-diisothiocyano-1,2-diphenylethane-2,2'-disulfonic acid (3H2DIDS) at 4 degrees C leads to preferential labeling of the cell surface coat. We have combined studies of the kinetics of 3H2DIDS-label release, the effects of enzymatic treatment, and cell electrophoretic mobility to characterize the 3H2DIDS-labeled components of the cell surface. Approximately 73% of the cell-associated radioactivity is spontaneously released from the cells after 5 h at 23 degrees C. The kinetics of release is consistent with the first-order loss of two fractions; a slow (tau 1/2 = 360 min) component representing 33% of the radioactivity and a fast (tau 1/2 = 20 min) component representing 26%. The remaining 14% of the labile binding may reflect mechanically induced surface release. Trypsin (1 microgram/ml) also removes approximately 73% of the labeled material within 30 min and converts the kinetics of release to that of a single component (tau 1/2 = 5.5 min). The specific activity (SA) of material released by trypsin immediately after labeling is 83% of the SA of the material spontaneously lost in 1 h. However, trypsinization following a 2-h period of spontaneous release yields material of reduced (43%) SA. Neither 3H2DIDS labeling nor the initial spontaneous loss of labeled material alters cell electrophoretic mobility. However, extended spontaneous release is accompanied by a significant decrease in surface charge density. Trypsinization immediately following labeling or after spontaneous release (2 h) reduces mobility by 32%. We have tentatively identified the slowly released compartment as contributing to cell surface negativity.  相似文献   

6.
Propionic acid (PA) is widely used as additive in animal feed and also in the manufacturing of cellulose-based plastics, herbicides, and perfumes. Salts of propionic acid are used as preservative in food. PA is mainly produced by chemical synthesis. Nowadays, PA production by fermentation of low-cost industrial wastes or renewable sources has been an interesting alternative. In the present investigation, PA production by Propionibacterium acidipropionici ATCC 4965 was studied using a basal medium with sugarcane molasses (BMSM), glycerol or lactate (BML) in small batch fermentation at 30 and 36 degrees C. Bacterial growth was carried out under low dissolved oxygen concentration and without pH control. Results indicated that P. acidipropionici produced more biomass in BMSM than in other media at 30 degrees C (7.55 g l(-1)) as well as at 36 degrees C (3.71 g l(-1)). PA and biomass production were higher at 30 degrees C than at 36 degrees C in all cases studied. The best productivity was obtained by using BML (0.113 g l(-1) h(-1)), although the yielding of this metabolite was higher when using glycerol as carbon source (0.724 g g(-1)) because there was no detection of acetic acid. By the way, when using the other two carbon sources, acetic acid emerged as an undesirable by-product for further PA purification.  相似文献   

7.
In pH-controlled batch fermentations with pure sugar synthetic hardwood hemicellulose (1% [w/v] glucose and 4% xylose) and corn stover hydrolysate (8% glucose and 3.5% xylose) lacking acetic acid, the xyloseutilizing, tetracycline (Tc)-sensitive, genomically integrated variant of Zymomonas mobilis ATCC 39676 (designated strain C25) exhibited growth and fermentation performance that was inferior to National Renewable Energy Laboratory's first-generation, Tc-resistant, plasmid-bearing Zymomonas recombinants. With C25, xylose fermentation following glucose exhaustion wasmarkellyslower, and the ethanol yield (based on sugars consumed) was lower, owing primarily to an increase in lactic acid formation. There was an apparent increased sensitivity to acetic acid inhibition with C25 compared with recombinants 39676:pZB4L, CP4:pZB5, and ZM4:pZB5. However, strain C25 performed well in continous ferm entation with nutrient-rich synthetic corn stover medium over the dilution range 0.03–0.06/h, with a maximum provess ethanol yield at D=0.03/h of 0.46 g/g and a maximum ethanol productivity of 3 g/(L·h). With 0.35% (w/v) acetic acid in the medium, the process yield at D=0.04/h dropped to 0.32 g/g, and the maximum productivity decreased by 50% to 1.5 g/(L·h). Under the same operating conditions, rec Zm Zm 4:pZB5 performed better; however, the medium contained 20 mg/L of Tc to constantly maintain selective pressure. The absence of any need for antibiotics and antiboitic resistance genes makes the chromosomal integrant C25 more com patible with current regulatory specifications for biocatalysts in large-scale commercial operations.  相似文献   

8.
Anti-infectious lock is an emerging therapeutic option for preventing and/or controlling catheter-associated infection. Ethanol has widespread bactericidal activity, limited side effects, and low risk of inducing antimicrobial resistance. However, concerns have been raised about ethanol-induced catheter structural degradation. In this study, silicone catheters were immersed at 37 degrees C in three different solvents: 0.9% sodium chloride, 60% ethanol, and 95% ethanol for 4 h, 15 days and 15 days after a first storage of 4 h. Scanning electron microscopy (magnification 1000-20 000 times) of the inner surface of the catheter revealed no damage to the lumen surfaces of catheters immersed in 95% ethanol for 15 days compared with the reference catheter. Gas chromatography/mass spectrometry (GC/MS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) analysis of the storage solutions revealed a significant release of polydimethylsiloxanes having a number of dimethylsiloxane units lower than 30 in the 95% ethanol solution and a structure highly consistent with a cyclic structure. Most release occurred within the first 4 h of exposure. In contrast, there was no difference in the small amounts of silicone released in 0.9% sodium chloride as reference and 60% ethanol solution, whatever the exposure time. These results should allow the development of clinical trials to assess the efficacy of the 60% ethanol lock technique in preventing or controlling the infectious complications of silicone dialysis catheters.  相似文献   

9.
Proteases are some of the most important industrial enzymes, and one of their main applications is for the production of cheese in the dairy industry. Due to a shortage of animal rennet, microbial coagulant proteases are being sought. In this work, the production of microbial rennet from Thermomucor indicae-seudaticae N31 was studied in submerged fermentation. The best enzyme production was obtained in a fermentation medium containing 4 % wheat bran as the substrate in 0.3 % saline solution, incubated for 72 h at 45 °C and 150 rpm. The value of the milk clotting activity (MCA) was 60.5 U/mL, and the ratio to proteolytic activity (MCA/PA) was 510. The crude enzyme showed optimum pH at 5.5 and two peaks of optimum temperature (MCA at 65 °C and PA at 60 °C). The MCA was stable in the pH range 4.0–4.5 for 24 h and up to 55 °C for 1 h. It was stable during storage at different temperatures (?20 to 25 °C) for 10 weeks. Based on these results, we conclude that microbial rennet from T. indicae-seudaticae N31 produced by submerged fermentation showed good prospects of replacing traditional rennet.  相似文献   

10.
The stability of caffeic acid phenethyl ester (CAPE) and its fluorinated derivative (FCAPE) in rat plasma and conditions preventing their degradation are reported. Reverse-phase high-pressure liquid chromatography (HPLC) using taxifolin as an internal standard was applied for the quantitative determination of CAPE and FCAPE in rat plasma extracted with ethyl acetate. The assay was validated over a linear range of 0.25-10 microg/mL plasma (r(2) > 0.9990, n = 3). No endogenous interferences were observed in the chromatographic region of interest. The limits of quantification and detection were set at 0.25 and 0.1 microg/mL, respectively. The precision ranged from 0.7 to 13.7% for CAPE, and from 0.4 to 10.4% for FCAPE. Accuracy ranged from -2.8 to 12.4% for CAPE and from -0.6 to 6.8% for FCAPE. The stability was conducted at 4, 25 and 37 degrees C. First-order kinetics was observed for the degradation of CAPE and FCAPE. The energies of activation of CAPE and FCAPE were found to be 17.9 and 20.1 kcal/mol, respectively. Addition of 0.4% of sodium chloride and pH adjustment to 6 prevented their degradation in rat plasma for 24 h and at least one month at -20 degrees C. This study provides useful information for the future pharmacokinetic study of CAPE and FCAPE in rat.  相似文献   

11.
Tarasiewicz M  Basińska H 《Talanta》1974,21(6):425-430
In neutral medium, picric acid (PA) reacts with chlorpromazine (CPM) hydrochloride with the formation of an orange compound, sparingly soluble in water, of molar ratio PA:CPM = 1:1. The compound is thermally stable up to a temperature of about 200 degrees C. The m.p. (178 degrees C) can be used for identification of picric acid as well as of chlorpromazine hydrochloride. Chlorpromazine picrate is extracted quantitatively with benzene, and the extract can be used for colorimetric determination of chlorpromazine hydrochloride. Beer's law is obeyed in the CPM concentration range 10-60 mug/ml.  相似文献   

12.
The stability of aflatoxins B1, B2, G1, and G2 was studied in solutions containing different concentrations of water, acetonitrile, and/or methanol, and in autosampler vials treated with nitric acid or silanized. When stored at room temperature (20 degrees C) for 24 h, aflatoxins G1 and G2 were stable only in solutions containing 100% organic solvent, whereas aflatoxins B1 and B2 were stable in solutions of methanol-water and acetonitrile-water at greater than 60 and 40% organic content, respectively. At 5 degrees C, aflatoxins G1 and G2 showed a significant decrease in concentration only when kept in less than 20% aqueous organic solvent. Significant loss of aflatoxins was realized in standard, commercially available amber type I borosilicate autosampler vials, but chemical etching of the vials with nitric acid or with silanization prevented aflatoxin degradation. These results indicate that aflatoxins are unstable in aqueous solutions and that this instability can be counteracted by the presence of at least 20% organic solvent and keeping the solutions at 5 degrees C or by the use of treated vials.  相似文献   

13.
Ma H  Hua Y  Tu C  Yuan L  Wei P 《色谱》2012,30(3):304-308
以对甲氧基苯胺为衍生试剂,采用毛细管电泳法分析了藏红花植物细胞多糖中的单糖组成。对衍生条件进行了优化,并对毛细管分离条件进行了系统的研究。衍生反应在醋酸含量9.5%(v/v)、80 ℃下反应2 h的衍生产率最大,衍生产物紫外检测波长234 nm。在优化的毛细管电泳分离条件(未涂层熔融石英毛细管柱(60 cm(有效长度50 cm)×50 μm),柱温25 ℃,电压20 kV,使用350 mmol/L硼酸电解液(pH 10.21),压力(3.4475 kPa)进样5 s)下,基线分离了11种结构相近的醛糖(来苏糖、木糖、核糖、葡萄糖、甘露糖、半乳糖、鼠李糖、纤维二糖、麦芽糖、乳糖)、酮糖(果糖)的衍生产物。应用该方法定量检测了藏红花植物细胞多糖水解物中糖的成分,各糖的回收率为94.3%~105.4%,相对标准偏差为3.3%~4.6%。  相似文献   

14.
Bag teas, packed 3g of ground black, green, oolong, paochoung and pu-erh tea leaves (the particle size used was 1-2mm), were steeped in 150 mL of 70, 85 or 100 degrees C hot water to study the effects of the number of steeping (the same bag tea was steeped repeatedly eight times, 30s each time, as done in China for making ceremonial tea) and varied steeping durations (0.5-4 min) on caffeine, catechins and gallic acid in tea infusions. The changes in tea infusions during storage at 4 or 25 degrees C for 0-48 h and the variations in these compounds of bag tea infused with 150 mL of 4 or 25 degrees C cold water for 0.5-16 h were also investigated. A HPLC method with a C18 column and a step gradient solvent system consisting of acetonitrile and 0.9% acetic acid in deionized water was used for analysis. Results for all kinds of tea samples showed that the second tea infusion contained the highest contents of caffeine, catechins and gallic acid when bag teas were steeped in 70 degrees C water. It was different from that steeped at 85 and 100 degrees C, the highest contents existed in the first infusion. These compounds decreased gradually in later infusions. Higher amounts of caffeine, catechins and gallic acid could be released from bag teas as hotter water was used. As steeping duration prolonged, these ingredients increased progressively, however, their levels were lower than that cumulated from the infusions with the identical bag tea prepared recurrently at the same temperature and time points. (-)-Gallocatechin gallate and (+)-catechin existed in these tea infusions rarely and could not be detected until a certain amount of them infusing. Except gallic acid that showed a significant increase and caffeine that exhibited no significant change, all kinds of catechins decreased appreciably after tea infusions were stored at 25 degrees C for 36 h; nevertheless, all of them showed no evident changes at 4 degrees C storage. The caffeine, catechins and gallic acid in tea infused with cold water also increased with increasing duration. Their contents in 25 degrees C steeped tea were higher than that made at 4 degrees C; moreover, their infusion rates from bag teas to cold water were markedly lower than that steeped in hot water. Infusing efficiencies of non-gallated catechins were higher than gallated catechins under cold water steeping.  相似文献   

15.
The kinetics of adsorption by sodium, ammonium, calcium and magnesium forms of zeolite Y from aqueous solutions containing 25% w/v of either one or an equimolar mixture of glucose (G) and fructose (F) have been studied batch-wise at 50 degrees C. The adsorption of aqueous pure G was fast, while that of aqueous pure F depended on the cationic form, approaching that of G on the Mg-Y, and slowing down in the sequence of Mg2+ > NH4+ > Ca2+ > Na+ of the cations. The adsorption behavior from solutions containing both G and F indicated significant hindering effects of F on G on Na-Y. Na-Y and Mg-Y did not exhibit rate-based selectivity, while Ca-Y an NH4-Y adsorbed G faster than F. Addition of CaCl2 to the mixture of Ca-Y and aqueous solution of G and F improved the separation, by hindering the adsorption of F. Addition of NH4Cl to the mixture of the sugar solution and NH4-Y, on the other hand, had a negative effect on the separation. NH4-Y was found to be desorbing about 30% of the adsorbed sugars and this value was found to be around 50% for Ca-Y. Re-adsorption experiments resulted in similar or somewhat higher percentages of amounts adsorbed compared to adsorption on fresh samples. Both NH4-Y and Ca-Y were found to be re-adsorbing around 50% of the sugars they adsorbed on fresh samples.  相似文献   

16.
Deswelling kinetics of water and terbinafine hydrochloride adsorbed poly(N-vinyl-2-pyrrolidone/itaconic acid) P(VP/IA) hydrogels were investigated. Hydrogels were prepared by irradiating the ternary mixture of VP/IA and cross-linking agent ethylene glycol dimethacrylate (EGDMA) in water by γ-rays at ambient temperature. Hydrogels swelled in pure water and terbinafine hydrochloride (TER-HCl) solutions at room temperature and deswelling or water loss were investigated between 4 and 45 °C temperature range and on human skin. The influence of IA content, % swelling, temperature and TER-HCl content on the water loss from gel matrix were investigated. Induction time for 80% water loss from hydrogel systems are found to increase from 9.6 to 21.2 h by increasing IA content in the gel system at 25 °C and decreased by 11 h with addition of TER-HCl in the gel system. Kinetic analyses had shown that the basic properties affecting the water loss behavior of these hydrogels are the IA and TER-HCl content and temperature of the medium.  相似文献   

17.
The growth and dissolution rates of glucose isomerase crystals ({1 0 1} face) were measured in situ at 0.1 and 100 MPa. From these data, we determined that the solubilities at 25 degrees C were C(e) = 3.1 +/- 0.9 and 2.6 +/- 0.5 mg mL(-1) at 0.1 and 100 MPa, respectively. At the same supersaturation of sigma = 2.5 (sigma identical with ln(C/C(e)), C = the concentration of glucose isomerase, C(e) = the solubility) and temperature (T = 25 degrees C), the growth rate under 100 MPa was 7.6 times larger than that under 0.1 MPa. This result shows, for the first time, a kinetic acceleration of the growth rates of protein crystals with increasing pressure. The growth rates vs sigma data fitted well with a two-dimensional nucleation growth model of a polynucleation type. The fitting results indicate that the acceleration is mainly due to the decrease in the molecular surface energy of the glucose isomerase crystal with pressure.  相似文献   

18.
Oxidation of 3b or 14c,d with Pb(OAc)4 in AcOH for 20 min at 25 degrees C and 1 h at 75 degrees C gave flavoskyrin-type dimers 6b and 15c,d in 53-86% yield. Heating a solution of 6b or 15c,d in pyridine under air for 1 h at 75-80 degrees C and then for 1-2 h at 110 degrees C afforded rugulosin-type dimers 10b and 17c,d in 61-88% yield. This two-step sequence provides a practical route to this unusual natural product skeleton.  相似文献   

19.
Photoinactivation kinetics of photosystem I (PSI)-mediated electron transport rate was compared to that of P700 content at room (22 degrees C) and low (4 degrees C) temperatures in isolated spinach thylakoid membranes. The high light treatment was carried out under aerobic and anaerobic conditions. At 22 degrees C the decrease of electron transport rate showed first order exponential kinetics. The amount of P700 decreased linearly, being less affected in the first hours of illumination. During photoinhibition at 4 degrees C in the presence of oxygen, the kinetics of inactivation of PSI photochemical activity and the content of P700 were different. It was found that 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) had different protective effect on the electron transport rate and on P700 content at both temperatures. Treatment with high light intensity under N(2) atmosphere had no effect on the electron transport rate or P700 content. The possible degradation of PSI reaction centre proteins was determined using immunoblot methods. In the presence of linear electron transport at 22 degrees C correlation between formation of toxic hydroxyl radicals and inhibition of oxygen uptake was observed.  相似文献   

20.
The binding of H2O to MeAl(OAr)2 (1: Ar = 2,6-di-tert-butyl-4-methylphenyl) in THF-d8 at -40 degrees C provides aquo complex 2, the structure of which was determined by X-ray crystallography. Complex 2 is unstable above 0 degrees C in THF-d8 and decomposes to form ArOH (major), CH4 (minor), and a methyl aluminoxane of undetermined structure. Decomposition of 2 follows first-order kinetics with k = 3.0 x 10-4 s-1 at 5 degrees C. The hindered phenol ArOH slowly reacts with [Cp2ZrMe][MeB(C6F5)3] (4) in bromobenzene-d5 solution at 25 degrees C to furnish CH4 and [Cp2ZrOAr][MeB(C6F5)3] (5), the structure of which was confirmed by X-ray crystallography. This reaction follows second-order kinetics for [ArOH] = [4] = 0.045 M and with k = 2.8 x 10-3 M-1 s-1 at 25 degrees C. This corresponds to a rate that is >107 x slower than the apparent rate of ethylene insertion for 4 at 25 degrees C at typical concentrations encountered in olefin polymerization. The kinetic data, as well as control experiments involving the addition of ArOH to active catalyst producing poly(ethylene), demonstrate that ArOH has essentially no effect on polymerization kinetics involving 4.  相似文献   

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