共查询到20条相似文献,搜索用时 15 毫秒
1.
《Process Biochemistry》2010,45(11):1795-1799
Microbial production and isolation of biosurfactants was studied. The production of lipopeptides surfactin and fengycin was performed by free and immobilized aerobic cells of Bacillus subtilis ATCC 21332. After preliminary tests with 5 polymer materials, the particles of polypropylene foamed with powder activated carbon (PPch) were selected for lipopeptides production for their thermal and mechanical stability and for the high colonizing effect. To avoid foaming during biosurfactant production, biofilm grown on solid floating support was aerated by air injected over the surface of cultural medium. The synthesis of both lipopeptides and especially of the fengycin was greatly enhanced by the immobilization. The relationship between support wettability, colonization of the cells, and lipopeptide production was discussed. Extraction behaviour of the lipopeptides into alkanes was studied. The distribution ratio of surfactin was found to be higher than this of fengycin at the same conditions and the n-heptane was more efficient solvent for both lipopeptides. Kinetics of surfactin recovery from fermentation broth applying batch pertraction in a rotating discs contactor was studied. Lipopeptide was successfully extracted (more than 75% in the first hour) using n-heptane as liquid membrane and a 0.2 mol L−1 phosphate buffer solution (pH ∼ 7.3) as receiving solution. However, the stripping of the organic liquid and surfactin accumulation into the receiving phase were less efficient. 相似文献
2.
Elham Tahmasebi Yadollah Yamini Abolfazl Saleh 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2009,877(20-21):1923-1929
The applicability of hollow fiber liquid phase microextraction (HF-LPME) for extraction and preconcentration of trace amounts of pioglitazone (PGL) as an anti-diabetic drug in biological fluids, prior to determination by high-performance liquid chromatography (HPLC), was evaluated. In this technique, the target drug was extracted into di-n-hexyl ether immobilized in the wall pores of a porous hollow fiber from 10 mL of the aqueous sample (source phase, SP) with pH 8.0, and then back extracted into the receiving phase (RP) with pH 2.2 located in the lumen of the hollow fiber. The extraction occurred due to a pH gradient between the two sides of the hollow fiber. After extracting for a prescribed time, 24 μL of the RP solution was taken back into the syringe and injected directly into a HPLC instrument for quantification. The Taguchi orthogonal array (OAD) experimental design with an OA16 (45) matrix was employed to optimize the HF-LPME conditions. Different factors affecting the HF-LPME efficiency such as the nature of organic solvent used to impregnate the membrane, pH of the SP and RP, stirring speed, extraction time and ionic strength were studied and optimized. Under the optimum conditions (di-n-hexyl ether as membrane impregnation solvent, pHs of the SP and RP equal to 8.0 and 2.2, respectively, extraction time of 30 min, stirring speed of 500 rpm and 10% (w/v) NaCl for adjusting the ionic strength), preconcentration factor of 180, linear dynamic range (LDR) of 2.5–250 μg L?1 with good correlation of determination (r2 > 0.998) and limit of detection (LOD) of 1.0 μg L?1 were obtained for the target drug. The percent relative intra-day and inter-day standard deviations (RSDs%) based on five replicate determinations were 4.7 and 15%, respectively. Once LPME was optimized, the performance of the proposed technique was evaluated for the determination of PGL in different types of biological fluids such as plasma and urine samples. The results showed that the proposed HF-LPME method could be successfully applied to determine trace amounts of PGL in biological samples. 相似文献
3.
Lectin from crude extract of small black kidney bean (Phaseolus vulgaris) was successfully extracted using the reversed micellar extraction (RME). The effects of water content of organic phase (Wo), ionic strength, pH, Aerosol-OT (AOT) concentration and extraction time on the forward extraction and the pH and ionic strength in the backward extraction were studied to optimize the extraction efficiency and purification factor. Forward extraction of lectin was found to be maximum after 15 min of contact using 50 mM AOT in organic phase with Wo 27 and 10 mM citrate-phosphate buffer at pH 5.5 containing 100 mM NaCl in the aqueous phase. Lectin was backward extracted into a fresh aqueous phase using sodium-phosphate buffer (10 mM, pH 7.0) containing 500 mM KCl. The overall yield of the process was 53.28% for protein recovery and 8.2-fold for purification factor. The efficiency of the process was confirmed by gel electrophoresis analysis. 相似文献
4.
《Biochimica et Biophysica Acta - Proteins and Proteomics》2018,1866(2):373-378
In alkaline media (pH 12) a catalytic peroxidase activity of cytochrome b5 was found associated to a different conformational state. Upon incubation at this pH, cytochrome b5 electronic absorption spectrum was altered, with disappearance of characteristic bands of cytochrome b5 at pH 7.0. The appearance of new electronic absorption bands and EPR measurements support the formation of a cytochrome b5 class B hemichrome with an acquired ability to bind polar ligands. This hemichrome is characterized by a negative formal redox potential and the same folding properties than cytochrome b5 at pH 7. The acquired peroxidase-like activity of cytochrome b5 found at pH 12, driven by a hemichrome formation, suggests a role of this protein in peroxidation products propagation. 相似文献
5.
The effects of different oxygen vectors on the synthesis and molecular weight of poly(γ-glutamic acid) (PGA) were investigated in the batch fermentation of Bacillus subtilis NX-2. n-Hexane, n-heptane, and n-hexadecane enhanced the PGA concentration and molecular weight. The PGA concentration reached a maximum of 39.4 ± 0.19 g L?1, and the highest molecular weight obtained was (19.0 ± 0.02) × 105 Da with the addition of 0.3% n-heptane. However, n-dodecane decreased the PGA concentration and molecular weight to final values of 20.1 ± 0.10 g L?1 and (8.4 ± 0.02) × 105 Da, respectively. Analysis of the intracellular nucleotide levels of B. subtilis NX-2 with n-heptane and n-dodecane additives showed that the lowest NADH/NAD+ ratio and ATP levels were obtained with the n-dodecane additives, which can explain the decreased PGA yield and molecular weight. The metabolic flux distribution of B. subtilis NX-2 with n-heptane and n-dodecane additives was also investigated. Flux distribution was primarily directed to the EMP and TCA cycles with n-heptane additives. The flux of 2-oxoglutarate to intracellular glutamate and the flux distribution from extracellular to intracellular glutamate both increased to improve PGA production. 相似文献
6.
François Coutte Didier Lecouturier Valérie Leclère Max Béchet Philippe Jacques Pascal Dhulster 《Process Biochemistry》2013,48(1):25-32
Recently, a bubbleless membrane bioreactor (BMBR) has been successfully developed for biosurfactant production by Bacillus subtilis [1]. In this study, for the first time, continuous culture were carried out for the production of surfactin in a BMBR, both with or without a coupled microfiltration membrane. Results from continuous culture showed that a significant part of biomass was immobilized onto the air/liquid membrane contactor. Immobilized biomass activity onto the air/liquid membrane contactor was monitored using a respirometric analysis. Kinetics of growth, surfactin and primary metabolites production were investigated. Planktonic biomass, immobilized biomass and surfactin production and productivity obtained in batch culture (3 L) of 1.5 days of culture were 4.5 g DW, 1.3 g DW, 1.8 g and 17.4 mg L?1 h?1, respectively. In continuous culture without total cell recycling (TCR), the planktonic biomass was leached, but immobilized biomass reached a steady state at an estimated 6.6 g DW. 11.5 g of surfactin was produced after 3 days of culture, this gave an average surfactin productivity of 54.7 mg L?1 h?1 for the continuous culture, which presented a surfactin productivity of 30 mg L?1 h?1 at the steady state. TCR was then investigated for the continuous production, extraction and purification of surfactin using a coupled ultrafiltration step. In continuous culture with TCR at a dilution rate of 0.1 h?1, planktonic biomass, immobilized biomass, surfactin production and productivity reached 7.5 g DW, 5.5 g DW, 7.1 g and 41.6 mg L?1 h?1 respectively, after 2 days of culture. After this time, biomass and surfactin productions stopped. Increasing dilution rate to 0.2 h?1 led to the resumption of biomass and surfactin production and these values reached 11.1 g DW, 10.5 g DW, 7.9 g and 110.1 mg L?1 h?1, respectively, after 3 days of culture. This study has therefore shown that with this new integrated bioprocess, it was possible to continuously extract and purify several grams of biosurfactant, with purity up to 95%. 相似文献
7.
《Process Biochemistry》2014,49(7):1182-1188
An efficient method for removing microbial cells and macromolecular impurities and purification of surfaction from fermentation broth produced by Bacillus amyloliquefaciens fmb50 was carried out in this study. Among three inorganic flocculants and a macromolecular flocculants, the combination use of CaCl2 and Na2HPO4 was the most effective separation process. Addition of 50% ethanol into fermentation broth could not only disrupt the surfactin micells, but also promote the permeating of surfactin in filtration. The flocculation condition was optimized by an L9 (34) orthogonal design. The light transmittance, surfactin recovery rate, protein removal rate and filtration flux could reach to 96.3%, 95.31%, 56.59% and 3204.41 L m−2 h−1 respectively, the surfactin purity reached to 79.5% and the residual protein was 8.1% in separated product under the optimal flocculation condition (flocculants dosage 0.5%, pH 5.0, and temperature 35 °C). Validation test also demonstrated stable results under the optimal conditions. Due to higher efficiency, lower cost and scale-up more easily of flocculation and filtration processes, it is feasible to separate surfactin from fermentation broth. 相似文献
8.
Atsushi Mukaiyama Takashi Nakamura Koki Makabe Kosuke Maki Yuji Goto Kunihiro Kuwajima 《Journal of molecular biology》2013,425(2):257-272
The kinetic folding of β2-microglobulin from the acid-denatured state was investigated by interrupted-unfolding and interrupted-refolding experiments using stopped-flow double-jump techniques. In the interrupted unfolding, we first unfolded the protein by a pH jump from pH 7.5 to pH 2.0, and the kinetic refolding assay was carried out by the reverse pH jump by monitoring tryptophan fluorescence. Similarly, in the interrupted refolding, we first refolded the protein by a pH jump from pH 2.0 to pH 7.5 and used a guanidine hydrochloride (GdnHCl) concentration jump as well as the reverse pH jump as unfolding assays. Based on these experiments, the folding is represented by a parallel-pathway model, in which the molecule with the correct Pro32 cis isomer refolds rapidly with a rate constant of 5–6 s? 1, while the molecule with the Pro32 trans isomer refolds more slowly (pH 7.5 and 25 °C). At the last step of folding, the native-like trans conformer produced on the latter pathway isomerizes very slowly (0.001–0.002 s? 1) into the native cis conformer. In the GdnHCl-induced unfolding assays in the interrupted refolding, the native-like trans conformer unfolded remarkably faster than the native cis conformer, and the direct GdnHCl-induced unfolding was also biphasic, indicating that the native-like trans conformer is populated at a significant level under the native condition. The one-dimensional NMR and the real-time NMR experiments of refolding further indicated that the population of the trans conformer increases up to 7–9% under a more physiological condition (pH 7.5 and 37 °C). 相似文献
9.
Magda C. Semedo Amin Karmali Patrícia D. Barata José V. Prata 《Journal of Molecular Catalysis .B, Enzymatic》2010,62(1):96-103
The present work involves the use of p-tert-butylcalix[4,6,8]arene carboxylic acid derivatives (tButyl[4,6,8]CH2COOH) for selective extraction of hemoglobin. All three calixarenes extracted hemoglobin into the organic phase, exhibiting extraction parameters higher than 0.90. Evaluation of the solvent accessible positively charged amino acid side chains of hemoglobin (PDB entry 1XZ2) revealed that there are 8 arginine, 44 lysine and 30 histidine residues on the protein surface which may be involved in the interactions with the calixarene molecules. The hemoglobin–tButyl[6]CH2COOH complex had pseudoperoxidase activity which catalysed the oxidation of syringaldazine in the presence of hydrogen peroxide in organic medium containing chloroform. The effect of pH, protein and substrate concentrations on biocatalysis was investigated using the hemoglobin–tButyl[6]CH2COOH complex. This complex exhibited the highest specific activity of 9.92 × 10?2 U mg protein?1 at an initial pH of 7.5 in organic medium. Apparent kinetic parameters (V′max, K′m, k′cat and k′cat/K′m) for the pseudoperoxidase activity were determined in organic media for different pH values from a Michaelis–Menten plot. Furthermore, the stability of the protein–calixarene complex was investigated for different initial pH values and half-life (t1/2) values were obtained in the range of 1.96 and 2.64 days. Hemoglobin–calixarene complex present in organic medium was recovered in fresh aqueous solutions at alkaline pH, with a recovery of pseudoperoxidase activity of over 100%. These results strongly suggest that the use of calixarene derivatives is an alternative technique for protein extraction and solubilisation in organic media for biocatalysis. 相似文献
10.
Wang CN Chen JY Sabu S Chang YL Chang SD Kao CC Peng HH Chueh HY Chao AS Cheng PJ Lee YS Chi LM Wang TH 《Free radical biology & medicine》2011,50(9):1124-1130
In the complex mechanism of preeclampsia, oxidative stress is an important pathogenic factor, and F2-isoprostane is a marker of oxidative stress and lipid peroxidation. The objective of this study was to identify if the amniotic fluid (AF) levels of F2-isoprostanes were elevated in women who later developed preeclampsia. In this study, we analyzed AF F2-isoprostane concentrations with enzyme immunoassay (EIA), and the EIA results could be validated by quantitative mass spectrometry. The mean AF concentration of F2-isoprostanes was significantly higher in pregnancies with subsequent development of preeclampsia (123.1 ± 57.6 pg/ml, n = 85) than in controls (73.8 ± 36.6 pg/ml, n = 85). The AF elevation of F2-isoprostanes was even higher in the preeclampsia with intrauterine growth restriction group (138.3 ± 65.2 pg/ml, n = 39). The area under the curve of the receiver operating characteristics analysis for AF F2-isoprostanes assay was 0.81, supporting its potential as a biomarker for preeclampsia. These results indicate that oxidative stress existed before the onset of clinical preeclampsia, further suggesting that the elevation of AF F2-isoprostanes may be used as a guide for antioxidant supplementation to reduce the risk and/or severity of preeclampsia. 相似文献
11.
《生物化学与生物物理学报:生物膜》2015,1848(5):1127-1138
N,N-dimethyldodecylamine-N-oxide (C12NO) is a surfactant that may exist either in a neutral or cationic protonated form depending on the pH of aqueous solutions. Using small angle X-ray diffraction (SAXD) we observe the rich structural polymorphism of pH responsive complexes prepared due to DNA interaction with C12NO/dioleoylphosphatidylethanolamine (DOPE) vesicles and discuss it in view of utilizing the surfactant for the gene delivery vector of a pH sensitive system. In neutral solutions, the DNA uptake is low, and a lamellar Lα phase formed by C12NO/DOPE is prevailing in the complexes at 0.2 ≤ C12NO/DOPE < 0.6 mol/mol. A maximum of ~ 30% of the total DNA volume in the sample is bound in a condensed lamellar phase LαC at C12NO/DOPE = 1 mol/mol and pH 7.2. In acidic conditions, a condensed inverted hexagonal phase HIIC was observed at C12NO/DOPE = 0.2 mol/mol. Commensurate lattice parameters, aHC ≈ dLC, were detected at 0.3 ≤ C12NO/DOPE ≤ 0.4 mol/mol and pH = 4.9–6.4 suggesting that LαC and HIIC phases were epitaxially related. While at the same composition but pH ~ 7, the mixture forms a cubic phase (Pn3m) when the complexes were heated to 80 °C and cooled down to 20 °C. Finally, a large portion of the surfactant (C12NO/DOPE > 0.5) stabilizes the LαC phase in C12NO/DOPE/DNA complexes and the distance between DNA strands (dDNA) is modulated by the pH value. Both the composition and pH affect the DNA binding in the complexes reaching up to ~ 95% of the DNA total amount at acidic conditions. 相似文献
12.
Christiane Trevisan Slivinski Edgar Mallmann Janete Magali de Araújo David Alexander Mitchell Nadia Krieger 《Process Biochemistry》2012,47(12):1848-1855
We studied the production of surfactin by Bacillus pumilus UFPEDA 448 in solid-state fermentation (SSF), using a medium based on okara with the addition of sugarcane bagasse as a bulking agent. The optimum proportions of okara and sugarcane bagasse were 50% each, by mass. Due to the relatively high production of proteases during SSF, pre-hydrolysis of the okara with a protease did not improve surfactin levels. The optimum temperature for surfactin production was 37 °C, with the incubation temperature affecting the ratios of the various surfactin homologues produced. Cultivation in column bioreactors with forced aeration under optimized conditions gave a surfactin level of 809 mg L?1 of impregnating solution, which corresponds to 3.3 g kg-dry-solids?1. This is the highest surfactin level that has been produced to date in SSF with a non-recombinant microorganism. The peak O2 uptake rate was 20 mmol min?1 kg-initial-dry-solids?1, corresponding to metabolic waste heat production rate of 182 W kg-initial-dry-solids?1. The tensioactive properties of the surfactin were similar to those reported in the literature for surfactin produced by submerged fermentation. These results suggest that it might be feasible to use SSF to produce surfactin but at large scale special attention will need to be given to heat removal. 相似文献
13.
《Biochemical Engineering Journal》2000,4(3):189-195
The mass transfer characterization in reversed micellar extraction of amino acid phenylalanine (Phe) is presented. The mass transfer rates in forward extraction of Phe from aqueous KCl solutions (pH 1.4 – 2.3) to AOT/isooctane reversed micellar solutions and in backward extraction from the reversed micellar organic phase to KHCO3/KOH buffer solutions (pH 9.0 – 11.0) were investigated using a stirred cell with a flat liquid–liquid interface. Both the forward and the backward extraction rates are controlled by the interfacial rate processes, i.e., the solubilization and the release processes. The solubilizing rate constants for the forward extraction of Phe increase with decreasing pH and initial Phe concentration and with increasing initial AOT concentration. On the other hand, the releasing rate constants for the backward extraction decrease with increasing initial AOT concentration and with decreasing ionic strength, but are little influenced by pH. The backward extraction rates are fairly slow compared to the forward extraction rates, and are accelerated by the addition of 2-methyl-2-propanol, similar to the extraction of protein lysozyme. 相似文献
14.
《Process Biochemistry》2007,42(9):1319-1325
A two-step enzymatic resolution process for production of (R)- and (S)-glycidyl butyrate was investigated and the lipases were screened. The first step involved a hydrolysis of (R,S)-glycidyl butyrate catalyzed by porcine pancreatic lipase (S-favored) with an E of 21 for production of (R)-glycidyl butyrate (13.2 mmol, 98% ee, 36% yield) under the optimal conditions (pH 7.4, 30 °C, 30 mg/ml CTAB). Then, the recovered (R)-enriched glycidol (19.8 mmol, 65% ee, 56% yield) was used for transesterification catalyzed by Novozym 435 (R-favored) with an E of 69 to obtain (S)-glycidyl butyrate (15.1 mmol, 98% ee, 42% yield) under the optimum conditions (aW = 0.24, n-heptane, 80 min). 相似文献
15.
《Journal of plant physiology》2014,171(10):868-875
Gaseous nitrogen dioxide (NO2) can disturb normal plant growth and trigger complex physiological responses. NO2-induced responses are influenced by biotic or abiotic factors. In this study, we investigated the effects of exogenous sodium sulfide (Na2S, 5 mmol L−1) on epidermis and stomata related physico-chemical responses of hybrid poplar cuttings (Pouplus alba × P. berolinensis) to gaseous NO2 (4 μl 1−1) for three time periods (0, 14 and 48 h). We also investigated hydrogen sulfide (H2S), nitrate-nitrogen and nitrate reductase activity (NR) in control and Na2S treated plants. Our results showed that NO2 exposure for 48 h led to the decline of NR, maximal PSII quantum yield (Fv/Fm), net photosynthetic rate (Pn), and dark respiration rate (Rd). The maximum rate for the post-illumination carbon dioxide burst (PIB) occurred in 48-h exposed leaves 13–15 s after darkening. Moreover, NO2 exposure resulted in a significant increase in nitrogen percentage (from 0 to 33%) and a decrease in the macro and micro-elements of leaf surface. Spraying Na2S aqueous solution on the leaf surfaces significantly increased the thicknesses of palisade/spongy tissue and H2S content. Na2S pretreatment alleviated NO2-caused toxic effects as indicated by increased NR and higher values of Pn, Fv/Fm, and actual photochemical efficiency in light (ФPSII) compared with the control. Na2S pretreatment had no significant impacts on PIB-based photorespiration or elements composition of a leaf surface. 相似文献
16.
The action of 10 and 20 mM Ca against harmful Cd effect on PS2 complex isolated from leaves of Phaseolus coccineus L. cv. Pi?kny Ja? was studied. The changes in fast chlorophyll a fluorescence induction kinetics and protein composition of PS2 complex were the symptoms of Cd toxicity and Ca protection of PS2 complex. Calcium applied at 10 mM concentration prevented F0 reduction caused by the presence of 250–1000 μM Cd in the incubation mixture, but that of (the variable chlorophyll a fluorescence) Fv, Fm, Fv/F0, and Fv/Fm only at 250 μM Cd. Ca concentration doubling in the incubation mixture resulted in complete overcoming the toxicity of 250–1000 μM Cd to Fv and Fm. However, the protection of Fv/F0 and the photochemical efficiency of PS2 (Fv/Fm) from 1000 μM Cd was only partial even at 20 mM Ca. A protective effect of 10 mM Ca on D1, D2 and 17 kDa proteins was found in PS2 complex exposed to 250 μM Cd, and on 43 kDa protein in the complex treated with 500 μM Cd. However, 20 mM Ca counteracted the toxicity of 500 μM Cd to the 43, 47 and 17 kDa proteins, as well as the harmful effect of 1000 μM Cd on 47 and 17 kDa ones. 相似文献
17.
Small-molecule fluorescent sensors that allow specific measurement of nuclear pH in living cells will be valuable for biological research. Here we report that Hoechst-tagged fluorescein (hoeFL), which we previously developed as a green fluorescent DNA-staining probe, can be used for this purpose. Upon excitation at 405 nm, the hoeFL–DNA complex displayed two fluorescence bands around 460 nm and 520 nm corresponding to the Hoechst and fluorescein fluorescence, respectively. When pH was changed from 8.3 to 5.5, the fluorescence intensity ratio (F520/F460) significantly decreased, which allowed reliable pH measurement. Moreover, because hoeFL binds specifically to the genomic DNA in cells, it was applicable to visualize the intranuclear pH of nigericin-treated and intact living human cells by ratiometric fluorescence imaging. 相似文献
18.
Mahmoud A. Ibrahim Mona M. Mohamed Abdel-Hady M. Ghazy Hassan M.M. Masoud 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》2013,164(3):221-228
Three superoxide dismutases (EC 1.15.1.1) (TLSOD1, TLSOD2 and TLSOD3) were purified from larvae of the camel tick Hyalomma dromedarii by ammonium sulfate precipitation, ion exchange and gel filtration columns. SDS-PAGE revealed that the subunit molecular masses of the SODs are 40 ± 2 kDa, 67 ± 1.5 kDa and 45 ± 2.6 kDa for TLSOD1, TLSOD2 and TLSOD3, respectively. TLSOD1 and TLSOD2 are monomeric proteins, while TLSOD3 isoenzyme exhibits dimeric structure with native molecular mass of 90 kDa. The pI values are estimated at pH 8.0, pH 7.2 and pH 6.6 for the three SODs which displayed pH optima at 7.6, 8.0 and 7.8, respectively. CuCl2 and ZnCl2 increase the activity of TLSOD2 and TLSOD3, while MnCl2 increases the activity of TLSOD1. KCN inhibits the activity of TLSOD2 and TLSOD3, while a remarkable resistance of TLSOD1 isoenzyme was detected. TLSOD1 is suggested to be a manganese containing isoenzyme while TLSOD2 and TLSOD3 are suggested to be copper/zinc-containing isoenzymes. These results indicate the presence of three different forms of SODs in the larval stage of camel tick. This finding will contribute to our understanding of the physiology of these ectoparasites and the development of non-traditional methods to control them. 相似文献
19.
Mas E Barden AE Corcoran TB Phillips M Roberts LJ Mori TA 《Free radical biology & medicine》2011,50(9):1171-1176
General and spinal anesthesia are used extensively in orthopedic surgery. However, these methods of anesthesia may result in different amounts of oxygen being delivered to the patient. Ischemia/reperfusion injury after release of the tourniquet initiates free radical-mediated oxidative stress. F2-isoprostanes are reliable markers of in vivo lipid peroxidation. However, under conditions of high oxygen tension, isofurans are formed. We aimed to compare plasma isofurans and F2-isoprostanes in spinal versus general anesthesia in patients undergoing knee-replacement surgery in a randomized, blinded study. Thirty-nine patients were randomized to spinal (SA; n = 19) or general anesthesia (GA; n = 20). Blood was collected before anesthesia, and a tourniquet was then applied to the limb during surgery. After release of the tourniquet, blood samples were collected at 30 min, 2 h, and 24 h for measurement of plasma F2-isoprostanes and isofurans by gas chromatography–mass spectrometry. The two groups were comparable in age and body mass index. Plasma F2-isoprostanes were significantly lower in the GA patients compared with the SA patients (p = 0.045). In contrast, the GA patients had significantly elevated plasma isofurans (p = 0.032). Increased isofurans during GA compared with SA are likely to reflect increased oxidative stress due to elevated oxygen concentrations during GA. Further studies are required to assess the implications of these findings on perioperative outcomes. 相似文献
20.
M.D.G. Anderson S.A. Breidinger E.J. Woolf 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2009,877(11-12):1047-1056
HPLC–MS/MS methods for the determination of a Hepatitis C NS3/NS4 protease inhibitor (MK-7009) in human plasma and Tween-treated urine were developed and validated over the concentration range 1–1000 ng/mL and 0.2–100 μg/mL respectively. A stable isotope labeled internal standard (ISTD), D4-MK-7009, was employed. Analytes were chromatographed by reversed phase HPLC and quantified by an MS/MS system. Electrospray ionization in the positive mode was employed. Multiple reaction monitoring of the precursor to product ion pairs m/z 758.6 → 637.4 MK-7009 and m/z 762.5 → 637.4 ISTD was used for quantitation. Analyte and internal standard were extracted from 250 μL of plasma using an automated 96-well liquid–liquid extraction. Plasma pH adjustment prior to extraction minimized ionization suppression in plasma samples from patients with Hepatitis C. The urine method involved direct dilution in the 96-well format of 0.020 mL Tween-treated urine. These methods have supported several clinical studies. Incurred plasma sample reanalysis demonstrated adequate assay reproducibility and ruggedness. 相似文献