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1.
Carrageenans are widely utilized in many commercial applications such as the food and pharmaceutical industry, due to their excellent functional properties. In this study, a sensitive LC-MS/MS method was developed to determine κ-3, κ-5, and κ-7 carrageenan oligosaccharides simultaneously. Optimum MRM transitions for κ-3, κ-5, and κ-7 carrageenan oligosaccharides were(645.079→565.111, [M-H]-),(515.137→474.946, [M-2H]~(2-)), and(471.484→445, [M-3H]~(3-)), respectively. Chromatographic separation was performed on an Amide column coupled with a guard column operated at 60℃ under stepwise gradient elution. The linearity of the LC-MS/MS method for κ-3, κ-5, and κ-7 carrageenan oligosaccharides, evaluated over the concentration range of 0.10- 20.0 μmol L~(-1), was excellent. The precisions of the method for κ-3, κ-5, and κ-7 carrageenan oligosaccharides were from 0.91% to 9.66%, and the inter-day precisions were from 0.92% to 10.5%. Validation of the LC-MS/MS method indicated that the method was precise and in line with the CFDA guidance. This method has been successfully applied to an in vitro absorption study.  相似文献   

2.
The total content ot the nch amino acids in two common red algae, Potystpnoma urceolata and Polysiphonia japonica growing in the Qingdao seashore were determined. The algae powder was hydrolyzed by 6 mol/L HC1 at 110℃ for 48 h and determined by amino acid analyzer. The content was 25.35% and 24.16% , respectively, much higher than that of some other species. In addition, a nutritive liquid with abundant amino acids was prepared (by the enzymatic hydrolysis method using Polysiphonia urceolata ) as raw material for a kind of health beverage. The dried seaweed was decolored by 0.25% KMnO4 and 0.5% active carbon, then enzymalized. In the selection of enzymalizing condition, the orthogonal experimental design was used. Four factors including kinds of enzyme, quantity of enzyme, temperature and time were studied at 3 levels. According to the orthogonal design results, we can choose an optimal condition: hydrolyzing at 45℃ by neutral proteinase (0.25% , w/w) for 2 h, adjusting pH to 8.5, then adding trypsin (0.25% , w/w) and hydrolyzing for 2 h. Finally the above solution was alkalized by NaOH and neutralized by casein. After the hydrolyzed liquid was filtered and concentrated, suitable additives were added. The final products contain rich amino acids.  相似文献   

3.
We previously isolated a natural product, namely guaiazulene-2,9-dione showing strong antibacterial activity against Vibrio anguillarum, from a gorgonian Muriceides collaris collected in South China Sea. In this experiment, guaiazulene-2,9-dione was quantitatively synthesized with an optimized one-step bromine oxidation method using guaiazulene as the raw material. The key reaction condition including reaction time and temperature, drop rate of bromine, concentration of aqueous THF solution, respective molar ratio of guaiazulene to bromine and acetic acid, and concentration of guaiazulene in aqueous THF solution, were investigated individually at five levels each for optimization. Combined with the verification test to show the absolute yield of each optimization step, the final optimal condition was determined as： when a solution of 0.025mmol mL-1 guaiazulene in 80% aqueous THF was treated with four volumes of bromine at a drop rate of 0.1 mL min 1 and four volumes of acetic acid at -5℃ for three hours, the yield of guaiazulene-2,9-dione was 23.72%. This was the first report concerning optimized one-step synthesis to provide a convenient method for the large preparation of guaiazulene-2,9-dione.  相似文献   

4.
We previously isolated a natural product, namely guaiazulene-2,9-dione showing strong antibacterial activity against Vibrio anguillarum, from a gorgonian Muriceides collaris collected in South China Sea. In this experiment, guaiazulene-2,9-dione was quantitatively synthesized with an optimized one-step bromine oxidation method using guaiazulene as the raw material. The key reaction condition including reaction time and temperature, drop rate of bromine, concentration of aqueous THF solution, respective molar ratio of guaiazulene to bromine and acetic acid, and concentration of guaiazulene in aqueous THF solution, were investigated individually at five levels each for optimization. Combined with the verification test to show the absolute yield of each optimization step, the final optimal condition was determined as: when a solution of 0.025 mmol m L-1 guaiazulene in 80% aqueous THF was treated with four volumes of bromine at a drop rate of 0.1 m L min-1 and four volumes of acetic acid at-5℃ for three hours, the yield of guaiazulene-2,9-dione was 23.72%. This was the first report concerning optimized one-step synthesis to provide a convenient method for the large preparation of guaiazulene-2,9-dione.  相似文献   

5.
A new κ-carrageenase gene cgk S was cloned from marine bacterium Shewanella sp. Kz7 by using degenerate and site-finding PCR. The gene was comprised of an open reading frame of 1224 bp, encoding 407 amino acid residues, with a signal peptide of 24 residues. Based on the deduced amino acid sequence, the κ-carrageenase Cgk S was classified into the Glycoside Hydrolase family 16. The cgk S gene was expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity with a specific activity of 716.8 U mg-1 and a yield of 69%. Recombinant Cgk S was most active at 45℃ and p H 8.0. It was stable at p H 6.0–9.0 and below 30℃. The enzyme did not require Na Cl for activity, although its activity was enhanced by Na Cl. Cgk S degraded κ-carrageenan in an endo-fashion releasing tetrasaccharides and disaccharides as main hydrolysis products.  相似文献   

6.
This paper investigates a microwave heating method for the determination of chemical oxygen demand (COD) in seawater. The influences of microwave-power, heating time and standard substances on the results are studied. Using the proposed method, we analyzed the glucose standard solution, the coefficient of variation being less than 2%. Compared with the traditional electric stove heating method, the results of F-test and T-test showed that there was no significant difference between the two methods, but the microwave method had slightly higher precision and reproducibility than the electric stove method. With the microwave heating method, several seawater samples from Jiaozhou Bay and the South Yellow Sea were also analyzed. The recovery was between 97.5% and 104.3%. This new method has the advantages of shortening the heating time, improving the working efficiency and having simple operation and therefore can be used to analyze the COD in seawater.  相似文献   

7.
The effects of different hydrolysis methods on peptidoglycan(PG) were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp,Litopenaeus vannamei.PG derived from Bifidobacterium thermophilum was prepared in the laboratory and processed with lysozyme and protease under varying conditions to produce several different PG preparations.A standard shrimp feed was mixed with 0.05% PG preparations to produce a number of experimental diets for shrimp.The composition,concentration,and molecular weight ranges of the soluble PG were analyzed.Serum phenoloxidase and acid phosphatase activity in the shrimp were determined on Days 6-31 of the experiment.The protective activity of the PG preparations was evaluated by exposing shrimp to white spot syndrome virus(WSSV).Data on the composition of the PG preparations indicated that preparations hydrolyzed with lysozyme for 72 h had more low-molecular-weight PG than those treated for 24 h,and hydrolysis by protease enhanced efficiency of hydrolysis compared to lysozyme.SDS-PAGE showed changes in the molecular weight of the soluble PG produced by the different hydrolysis methods.Measurements of serum phenoloxidase and acid phosphatase activity levels in the shrimp indicated that the PG preparations processed with enzymes were superior to the preparation which had not undergone hydrolysis in enhancing the activity of the two serum enzymes.In addition,the preparation containing more low-molecular-weight PG enhanced the resistance of the shrimp to WSSV,whereas no increased resistance was observed for preparations containing less low-molecular-weight PG.These findings suggest that the immunity-enhancing activity of PG is related to its molecular weight and that increasing the quantity of low-molecular-weight PG can fortify the effect of immunity enhancement.  相似文献   

8.
The surface of ZrO2 nanoparticles was modified by styrene coupling grafting method to improve the dispersion and interaction of the nanoparticles with the epoxy coating in which the modified ZrO2 nanoparticles were used as an additive. The grafting performance and microstructure of the nano- ZrO2/epoxy coating were analyzed by Fourier transformation infrared spectroscopy, X-ray diffraction, and scanning electron microscopy. The corrosion behavior of the nano-ZrO2/epoxy coating on mild steel was evaluated in neutral 3.5 wt% NaC1 solution using electrochemical impedance spectroscopy （EIS）. Both the coating capacitance and coating resistance fitted by the equivalent circuit from EIS were used to evaluate the protective performance of the coating towards the mild steel. The results show a superior stability and efficient corrosion protection by the modified ZrO2 nanoparticles. The epoxy coating containing 2 wt% modified ZrO2 nanoparticles exhibited the best corrosion performance among all the coating specimens. This research may provide an insight into the protection of mild steel using modified epoxy coatings.  相似文献   

9.
The 60% ethanolic extract from Gracilaria textorii (Rhbdophyta) was degraded with B- agarase, and certain charged (sulfated) and neutral oligosaccharides were separated by using DEAE Sephadex A 25 and Bio- gel P6 , P2 chromatographic techniques . Some of the charged oligomers were verified to be neoagarotetraose - 63- sulfate ( DP2 ). neoagarohexaose- 63 > 65 - disulfate ( DP3 ) and neoagarooctaose- 63, 65, 67-trisulfate (DP4) by using 13C-and 'H-NMR spectroscopy . One neutral oligomer was assumed to be a mixture of methylated neoagarotetraoses (DP2 ) by 1H-NMR spectroscopy .These oligomers assigned by their chemical shifts may be used as the model compounds for the structural investigation of the agar- type sulfated polysaccharides using the B- agarase degradation method .  相似文献   

10.
A new kinetic spectrophotometric method has been developed for the determination of vanadium (Ⅴ).The method is based on the catalytic effect of vanadium (Ⅴ) on the oxidation of weak acid brilliant blue dye (RAWL) by KBrO3 using the citric acid as activation reagent.The obtained optimum conditions are:c (RAWL)=1×10-4 molL-1, c (KBrO3)=3×10-2 molL-1, c (citric acid)= 9×10-3 molL-1, pH=2.50, the reaction time being 7.0 min and the temperature being 25.0℃.Under the optimum conditions, the pro-posed method allows the determination of vanadium (Ⅴ) in the range of 0?70.0 ng mL-1 and the detection limit is down to 0.407 ng mL-1.For standard vanadium (Ⅴ) solution determination, the recovery efficiency is in the range of 98.5%?102% and the RSD ranges from 0.76%?1.25%.Moreover, it is demonstrated that most cations and anions do not interfere with the determination of vanadium (Ⅴ) under the analytical condition.The new method was successfully applied in the determination of vanadium (Ⅴ) in fresh water and seawater samples with satisfactory results.Vanadium (Ⅴ) in the seawater samples from Qingdao offshore was determined using the method and the distribution of vanadium (Ⅴ) was mapped.Compared with other instrumental analytical methods, the proposed method shows good selectivity, sensitivity, simplicity, lower cost and rapidity.It can be employed on shipboard easily.  相似文献   

11.
A highly sensitive method was developed for the simultaneous determination of 8 steroid hormones in high-fat fish tissues using ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).The 8 steroid hormones were extracted from the tissues with diethyl ether.Differing from other common purification methods,the extract solutions were cleaned by gel permeation chromatography(GPC) using ethyl acetate-cyclohexane solution(1:1,v/v) as the mobile phase.The separation of target compounds was carried out by a BEH C18 column and a gradient elution consisting of acetonitrile and 0.2% aqueous formic acid(v/v).The compounds were detected under the multiple reaction monitoring(MRM) mode and quantified with external standard method.This method was validated with respect to linearity,specificity,accuracy and precision.A linearity with correlation coefficient larger than 0.995 was achieved in the range of 0.5 to 50 ng m L~(-1).The average recoveries at the spiked levels of 1.0,5.0,and 10.0 μg kg~(–1) varied between 81.7% and 90.8%,with the relative standard deviations(n=5) ranged from 3.50% to 10.0%.The limit of quantification(LOQ) for 8 steroid hormones ranged from 0.2 to 1.5 μg kg~(-1).It was concluded that this method can be successfully applied for the determination of 8 steroid hormones in complicated matrices including high-fat fish tissues.  相似文献   

12.
This study gives an analytical solution for wave interaction with a partially reflecting vertical wall protected by a submerged porous bar based on linear potential theory. The whole study domain is divided into multiple sub-regions in relation to the structures. The velocity potential in each sub-region is written as a series solution by the separation of variables. A partially reflecting boundary condition is used to describe the partial reflection of a vertical wall. Unknown expansion coefficients in the series solutions are determined by matching velocity potentials among different sub-regions. The analytical solution is verified by an independently developed multi-domain boundary element method(BEM) solution and experimental data. The wave run-up and wave force on the partially reflecting vertical wall are estimated and examined, which can be effectively reduced by the submerged porous bar. The horizontal space between the vertical wall and the submerged porous bar is a key factor, which affects the sheltering function of the porous bar. The wave resonance between the porous bar and the vertical wall may disappear when the vertical wall has a low reflection coefficient. The present analytical solution may be used to determine the optimum parameters of structures at a preliminary engineering design stage.  相似文献   

13.
κ-carrageenan oligosaccharides exhibit various biological activities. Enzymatic degradation by κ-carrageenase is safe and controllable. Therefore, κ-carrageenases have captured more and more attentions. In this study, a κ-carrageenase encoding gene, cgk X, was cloned from Pseudoalteromonas sp. QY203 with degenerate and inverse PCR. It comprised an ORF of 1194 bp in length, encoding a protein with 397 amino acid residues. Cgk X is a new member of glycoside hydrolase family 16. The deduced amino acid sequence shared a high similarity with Cgk X of Pseudoalteromonas κ-carrageenase; however, the recombinant Cgk X showed different biochemical characteristics. The recombinant enzyme was most active at p H 7.0 and 55℃ in the presence of 300 mmol L~(~(-1))Na Cl. It was stable in a broad range of acidity ranging from p H 3.0 to p H 10.0 when temperature was below 40℃. More than 80% of its activity was maintained after being incubated at p H 3.6–10.0 and 4℃ for 24 h. Cgk X retained more than 90% of activity after being incubated at 40℃ for 1 h. EDTA and SDS(1 mmol L~(-1)) did not inhibit its activity. Cgk X hydrolyzed κ-carrageenan into disaccharide and tetrasaccharide as an endo-cleaver. All these characteristics demonstrated that Cgk X is applicable to both κ-carrageenan oligosaccharide production and κ-carrageenase structure-function research.  相似文献   

14.
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15.
A novel method for on-site determination of trace iron was developed using membrane preconcentration and spectrophotometric detection. Fe(Ⅱ)-ferrozine complex was reacted with cetyltrimethylammonium bromide (CTAB) to form a Fe(Ⅱ)-ferrozine-CTAB paired compound, which was collected on a membrane by filtration under vacuum. The membrane was immersed in 2 mL of ethanol-nitric acid and the absorbance of the solution measured for quantitative analysis. Various factors affecting the iron collection and determination were investigated. With different sample preconcentration volumes, the range of determination was broadened to 0.5-120 g/L. The detection limit of this method reached 0.19 g/L and the recoveries were between 97.2 and 109% when the concentration enrichment was about 45. The relative standard deviation (n = 7) was 1.9% for samples containing 10 g/L Fe. Twelve seawater samples were analyzed on-site using the proposed method, and two were also analyzed in inductively coupled plasma mass spectrometry. No significant difference was shown between the two methods by the Student’s t-test. The method has also been used on-site for iron enrichment experiments with phytoplankton and concluded to be simple, accurate and inexpensive.  相似文献   

16.
Vitellin of mature female marine crab Charybdis japonica was purified by high-performance liquid chromatography （HPLC, DEAE-cellulose-16 anion exchange column）. The apparent molecular mass of the vitellin is 546 ku based on the data of native-PAGE. Under denatured condition （SDS-PAGE）, it was found that vitellin was composed of four polypeptides each at 120, 100, 65, and 55 ku. One disulfide bond was detected in the binding of polypeptide subunits. The purified vitellin, contained 4.47% phosphor and 10.6% polysaccharides, and was identified as glyco-lipo-carotenoprotein, according to the PAGE staining data. The purified vitellin can be used as antigen to raise polyclonal antisera in further application.  相似文献   

17.
We have reported that A. pullulans 98 produces a high yield of cellulase. In this study, a carboxymethyl cellulase(CMCase) in the supernatant of the culture of A. pullulans 98 was purified to homogeneity, and the maximum production of CMCase was 4.51 U(mg protein)-1. The SDS-PAGE analysis showed that the molecular mass of the purified CMCase was 67.0 k Da. The optimal temperature of the purified enzyme with considerable thermosensitivity was 40℃, much lower than that of the CMCases from other fungi. The optimal p H of the enzyme was 5.6, and the activity profile was stable in a range of acidity(p H 5.0–6.0). The enzyme was activated by Na+, Mg2+, Ca2+, K+, Fe2+ and Cu2+, however, it was inhibited by Fe3+, Ba2+, Zn2+, Mn2+ and Ag+. Km and Vmax values of the purified enzyme were 4.7 mg m L-1 and 0.57 μmol L-1 min-1(mg protein)-1, respectively. Only oligosaccharides with different sizes were released from carboxymethylcellulose(CMC) after hydrolysis with the purified CMCase. The putative gene encoding CMCase was cloned from A. pullulans 98, which contained an open reading frame of 954 bp(EU978473). The protein deduced contained the conserved domain of cellulase superfamily(glucosyl hydrolase family 5). The N-terminal amino acid sequence of the purified CMCase was M-A-P-H-A-E-P-Q-S-Q-T-T-E-Q-T-S-S-G-Q-F, which was consistent with that deduced from the cloned gene. This suggested that the purified CMCase was indeed encoded by the cloned CMCase gene in this yeast.  相似文献   

18.
The anodic stripping voltammetry with physically-coating mercury film electrode was used to investigate the complexing action of trace heavy metals in seawater with organic ligands. The apparent copper complexing capacity of seawater was determined by titrating the organic ligands in natural seawater with standard ionic copper solution. The complexing actions of copper in seawater with humic acid (HA) or fulvic acid (FA) were investigated by titrating copper in seawater with HA or FA solution. The equilibrium time, electrodeposition potential, and effect of pH etc. were investigated respectively. The results show that the interaction of copper in seawater with organic matter is a fast process. At natural pH, HA or FA tend to act with copper in seawater to form non-labile complexes. During experimental electrode process, these complexes did not significantly dissociate. The experimental results were calculated according to 1:1 complex formation.  相似文献   

19.
The new polyenzyme method for making gravy from Harengula zunasi offal involves protein enzymolysis with flavorase after proper alkaline and neutral protease levels were established by orthogonal trials to select the best hydrolytic conditions for processing offal with alkaline and neutral protease. The conditions for the polyenzyme method were pH of 7.0, temperature of 50℃ , alkaline and neutral protease concentration of 1.5% respectively, hydrolysis time of 120 min, andflavorase concentration of 2.0% , for 60 min. The new gravy-making technology yields a nutritious and delicious gravy containing 40.3 % of total essential amino acids, with delicious amino acids Glu, Asp, Gly, Ala, Pro and Ser comprising 49.5% , total and amino nitrogen being respectively1.9 and 1.1 g/100 g (amino acid nitrogen being 61.0% of total nitrogen), The polyenzyme method was used to make 14.8% protein gravy from Harengula zunasi offal. In addition, in inorganic elements, the phosphorus content is the highest.  相似文献   

20.
Ultrasound-assisted solvent extraction was confirmed as a novel,effective method for separating lipid from mackerel pro-tein,resulting in a degreasing rate (DR) of 95% and a nitrogen recovery (NR) of 88.6%.To obtain protein hydrolysates with high ni-trogen recovery and low bitter taste,enzymatic hydrolysis was performed using eight commercially available proteases.It turned out that the optimum enzyme was the ‘Mixed enzymes for animal proteolysis’.An enzyme dosage of 4%,a temperature of 50℃,and a hydrolysis time of 300 min were found to be the optimum conditions to obtain high NR (84.28%) and degree of hydrolysis (DH,16.18%) by orthogonal experiments.Glutamic acid was the most abundant amino acid of MDP (defatted mackerel protein) and MDPH (defatted mackerel protein hydrolysates).Compared with the FAO/WHO reference protein,the essential amino acid chemical scores (CS) were greater than 1.0 (1.0 1.7) in MDPH,which is reflective of high nutritional value.This,coupled with the light color and slight fishy odor,indicates that MDPH would potentially have a wide range of applications such as nutritional additives,functional ingredients,and so on.  相似文献   

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