Feasibility of pressurization to speed up enzymatic hydrolysis of biological materials for multielement determinations

The feasibility of pressurized solvents (liquids at a high pressure and/or high temperature without the subcritical point being reached) has been newly investigated to accelerate enzymatic hydrolysis processes of mussel tissue for multielement determinations. The target elements (Al, As, Cd, Co, Cu, Fe, Hg, Li, Mn, Pb, Se, Sr, V, and Zn) were released from dried mussel tissue by action of two proteases (pepsin and pancreatin), and they have been evaluated by inductively coupled plasma optical emission spectrometry (ICP-OES). Variables inherent to the enzymatic activity (pH, ionic strength, temperature, and enzyme mass) and factors affecting pressurization (static time, pressure, and number of cycles) were simultaneously studied by applying a Plackett-Burman design (PBD) as the screening method. Results showed that pH, ionic strength, and temperature were the most statistically significant factors (confidence interval of 95%) under pressurized conditions for pepsin, while pH and ionic strength affected pancreatin activity. This means that metal extraction is mostly attributed to enzymatic activity. The static time (enzymatic hydrolysis time) was found statistically nonsignificant for most of the elements, meaning that the hydrolysis procedure can be finished within a 2-15 min range. For pepsin, optimized conditions (pH 1.0, temperature 40 degrees C, pressure 1500 psi, static time 2 min, and number of cycles 3) gave quantitative extractions for As, Cd, Co, Cu, Hg, Li, Mn, Pb, Se, Sr, V, and Zn. The pepsin mass was 0.05 g, and the solution was Milli-Q water at pH 1.0 (adjusted with hydrochloric acid). For pancreatin, quantitative recoveries were only reached for As, Cd, Cu, Li, Pb, and Sr at room temperature, at a pressure of 1500 psi, for a static time of 2 min and a number of cycles of 3. The extraction solution was a 0.3 M potassium dihydrogen phosphate/potassium hydrogen phosphate buffer at a pH of 7.5 working at room temperature. Around 0.5 g of diatomaceous earth was used as dispersing agent for hydrolyses with either enzyme. Analytical performances, such as limits of detection and quantification and repeatability of the overall procedure, have been established. Finally, accuracy of the methods was assessed by analyzing seafood certified reference materials (GBW-08571, DORM-2, DOLT-3, TORT-2), fatty tissues certified reference materials (BCR 185, NIST 1577b), and fibrous certified reference materials (BCR 62, GBW-08501).     

A Standard for the Measurement of the pH of Blood and Other Physiological Media

A buffer solution containing potassium dihydrogen phosphate (0.008695 molal) and disodium hydrogen phosphate (0.03043 molal) is proposed as a pH standard for the physiologically important range, pH 7 to 8. The proposed standard solution is prepared by dissolving 1.179 g (air weight) of potassium dihydrogen phosphate and 4.303 g (air weight) of disodium hydrogen phosphate in ammonia-free water and diluting to 1 liter at 25 °C. The ionic strength is 0.1.Standard pH values (pH_s) were assigned to this reference solution at temperatures from 0 to 50 °C by means of emf measurements of hydrogen-silver chloride cells without liquid junction. The activity coefficient of chloride ion, upon which the assignment of pH_s depends, was evaluated by means of a recently proposed convention. By this means, standard values precise to ± 0.001 unit could be derived from the emf data. At 25 °C pH_s is 7.414, and at 38 °C it is 7.382. The operational consistency of these standard values with those for the 0.025-m equimolal phosphate buffer (one of the NBS primary standards) was demonstrated.     

Improvements on enzymatic hydrolysis of human hair for illicit drug determination by gas chromatography/mass spectrometry

The use of ultrasound energy for accelerating the pronase E enzymatic hydrolysis of human hair for extracting illicit drugs has been novelty tested. The enzymatic extracts obtained after 30 min of sonication in an ultrasonic water bath were subjected to an optimized solid-phase extraction process, which involved a solution of 2.0% (v/v) acetic acid in methanol as eluting solution and concentration by N2 stream evaporation. A gas chromatography/mass spectrometry method was used to separate and determine cocaine, benzoylecgonine, codeine, morphine, and 6-monoacethylmorphine in 20 min. Variables affecting ultrasound-assisted pronase E hydrolysis such as hydrolysis temperature, hydrolysis time, enzyme concentration, catalyzer (1,4-dithiothreitol) concentration, ionic strength, pH, and ultrasound frequency were simultaneously evaluated by a Plackett-Burman design 2(8) PBD of resolution III. The most statistically significant variables were ionic strength and pH, which means that analyte extraction is mainly attributed to pronase E activity. The optimization or evaluation of all the factors has led to an accelerated pronase E hydrolysis of human hair, which can be completed in 30 min. Results have been found to be statistically similar to those obtained with conventional pronase E hydrolysis. The accelerated method was finally applied to several human hair samples from multidrug abusers.     

Use of potassium dihydrogen phosphate and sawdust as adsorbents of ammoniacal nitrogen in aerobic composting process

Three kinds of adsorbents-potassium dihydrogen phosphate, sawdust and mixture of potassium dihydrogen phosphate and sawdust were added respectively into composting to investigate their adsorption effect on ammonia. The experimental results showed that all the adsorbents could restrain ammonia volatilizing, with the sorption of potassium dihydrogen phosphate adsorbents being the best of all, the sorption of mixture adsorbent with potassium dihydrogen phosphate and sawdust being the second and the sorption of sawdust adsorbent being the last. Therefore, the total nitrogen loss ratios respectively reduced from 38% to 13%, 15% and 21% after adding these three kinds of adsorbents into composting. However, potassium dihydrogen phosphate produced negative influence on composting properties as its supplemented amount exceeded a quantity basis equivalent to 18% of total nitrogen in the composting, for example: pH value had been lessened, microorganism activity reduced, which finally resulted in the reduction of biodegradation ratio of organic matter. But it did not result in these problems when using the mixture of potassium dihydrogen phosphate and sawdust as adsorbent, in which the amount of potassium dihydrogen phosphate was under a quantity basis equivalent to 6% of total nitrogen in the composting. Moreover, the mixture adsorbent produced better adsorption effect on ammonia, and raised biodegradation ratio of organic matter from 26% to 33%.     

Electrochemical production of Palladium and its use as indicator electrode

The electrodeposition of palladium from electrolytes containing palladium chloride along with potassium-dihydrogen phosphate, sodium fluoride or oxalic acid was studied under a variety of condition. The conditions used were: 1.0 M palladium chloride, 0.15 M potassium dihydrogen phosphate, pH 1.5 at 298 K and current density 0.85 mA cm^?2/ The current efficiency was calculated and the crystal structure confirmed by X-ray analysis. Spectrophotomteric and chemical analysis reveals that the purity of the metal is better than 99%. Palladium electrode can function satisfactorily as indicator electrode for hydrogen ions in acid-base titrations. Titration of acids with sodium hydroxide and of salts of weak acids with hydrochloric acid showed that palladium can indicate very accurately the end point. The end point coincide to great extent with the visual values with an error not more than ± 2%.     

Proton transport in KDP-family of ferroelectric materials

Proton transport in potassium dihydrogen phosphate (KDP) and ammonium dihydrogen phosphate (ADP) is briefly reviewed. The experimental results of Wagner’s polarization measurement, coulometry, infrared spectroscopy, transient ionic current measurement, the variation of electrical conductivity with temperature and electrogravimetric analysis for KDP and ADP are reported. H⁺ and OH^- ions are ascertained as the mobile ionic species. A new mechanism for the proton transport in KDP and ADP is suggested: A three-fold rotation of H₂P₄ ^- units about any of the axes of PO₄ tetrahedra leads to a configuration like O...H...H...O, the rearrangement of which provides H...O...H bridge that gets electrodissociated on the application of a d.c. electric field.     

Linear electrooptic effect in doped KDP crystals

The dispersion of half wave voltages with wave length and temperature of potassium dihydrogen ortho phosphate (KDP) crystals with additives and dopants like borax, nickel phosphate, manganese phosphate, sodium dihydrogen orthophosphate and potassium arsenate were determined. The refractive index for all the above crystals were found by the modified Rayleigh’s refractometer method. The linear electrooptic coefficients were calculated at various wavelengths of light. The variation of half wave voltage was also studied as a function of concentration of the dopant.     

The hydrolysis kinetics of monobasic and dibasic aminoalkyl esters of ketorolac

Six aminoethyl and aminobutyl esters of ketorolac containing 1-methylpiperazine (MPE and MPB), N-acetylpiperazine (APE and APB) or morpholine (ME and MB), were synthesized and their hydrolysis kinetics were studied. The hydrolysis was studied at pH 1 to 9 (for MPE, APE and ME) and pH 1 to 8 (for MPB, APB and MB) in aqueous phosphate buffer (0.16?M) with ionic strength (0.5?M) at 37°C. Calculation of k_obs, construction of the pH-rate profiles and determination of the rate equations were performed using KaleidaGraph^® 4.1. The hydrolysis displays pseudo-first order kinetics and the pH-rate profiles shows that the aminobutyl esters, MPE, APB and MB, are the most stable. The hydrolysis of the ethyl esters MPE, APE and ME, depending on the pH, is either fast and catalyzed by the hydroxide anion or slow and uncatalyzed for the diprotonated, monoprotonated and nonprotonated forms. The hydrolysis of the butyl esters showed a similar profile, albeit it was also catalyzed by hydronium cation. In addition, the hydroxide anion is 105 more effective in catalyzing the hydrolysis than the hydronium cation. The hydrolysis pattern of the aminoethyl esters is affected by the number and pKa of its basic nitrogen atoms. The monobasic APE and ME, show a similar hydrolysis pattern that is different than the dibasic MPE. The length of the side chain and the pKa of the basic nitrogen atoms in the aminoethyl moiety affect the mechanism of hydrolysis as the extent of protonation at a given pH is directly related to the pKa.     

钙系磷酸盐化学键合材料的制备及其固化重金属研究

利用富含氧化钙的铬铁渣(FS)和磷酸二氢钾(P)反应制备钙系磷酸盐化学键合材料,并用其作为固化重金属离子(Pb~(2+)、Cd~(2+)、Cu~(2+))基体材料。研究了原料配比、缓凝剂及重金属掺量对胶凝材料初凝时间和抗压强度的影响。结果表明:当P/FS(质量比,下同)为1/4及硼砂掺量为2%时,材料性能最好,自然养护28d和常压蒸汽养护24h抗压强度分别可达25.65 MPa和36.86 MPa。随着重金属掺量的增加,材料抗压强度逐渐降低,掺量为3%时,自然养护28d和蒸汽养护24h试块抗压强度均大于10 MPa,满足建筑材料要求。固化体重金属毒性浸出试验表明:磷酸盐化学键合材料对重金属离子(Pb~(2+)、Cd~(2+)、Cu~(2+))均具有很好的固化效果,浸出浓度远低于相应的鉴别标准。通过XRD、SEM和FTIR分析,钙系磷酸盐化学键合材料固化重金属的机理是通过水化产物的化学键合、吸附以及物理包裹作用将Pb~(2+)、Cd~(2+)、Cu~(2+)固化在材料中。     

Degradation kinetics of somatostatin in aqueous solution

The degradation kinetics of somatostatin (somatotropin release inhibiting factor), a cyclic tetradecapeptide, was investigated as a function of temperature, pH, ionic strength, buffer type, and buffer concentration. In addition, the effect of different container materials in which the solutions were stored and the presence of an antimicrobial agent for in vitro use was examined. The degradation of somatostatin followed first-order kinetics under all investigated conditions. The pH-stability profile showed a well-defined stability optimum around pH 3.7. The degradation was accelerated at higher buffer concentrations, phosphate buffer being significantly more detrimental than acetate buffer. The ionic strength and the drug concentration had virtually no effect on the degradation rate. When general purpose glass vials were used as storage containers, degradation was faster due to release of alkali from the container material. The solution properties, i.e., pH, buffer type, buffer capacity, and the experimental setup such as container material and sterile conditions need to be carefully selected or maintained, in order to avoid accelerated degradation.     

Self-Assembled Carbohydrate-Stabilized Ceramic Nanoparticles for the Parenteral Delivery of Insulin

The insulin-bearing aquasomes were fabricated by first preparing the nanosize calcium phosphate dihydrate core. The calcium phosphate dihydrate core was prepared by colloidal precipitation and sonication of disodium hydrogen phosphate solution and calcium chloride solution at low temperature. This core was coated with cellobiose, pyridoxal-5-phosphate, or trehalose under sonication and was further loaded with the drug at low temperature by a partial adsorption mechanism. The prepared systems were characterized for size, shape, size distribution, drug loading efficiency, and in vivo performance. The in vivo performance of the formulated aquasome was compared with standard porcine insulin solution, and better results were observed compared to insulin solution.     

Self-assembled carbohydrate-stabilized ceramic nanoparticles for the parenteral delivery of insulin

The insulin-bearing aquasomes were fabricated by first preparing the nanosize calcium phosphate dihydrate core. The calcium phosphate dihydrate core was prepared by colloidal precipitation and sonication of disodium hydrogen phosphate solution and calcium chloride solution at low temperature. This core was coated with cellobiose, pyridoxal-5-phosphate, or trehalose under sonication and was further loaded with the drug at low temperature by a partial adsorption mechanism. The prepared systems were characterized for size, shape, size distribution, drug loading efficiency, and in vivo performance. The in vivo performance of the formulated aquasome was compared with standard porcine insulin solution, and better results were observed compared to insulin solution.     

Stability of Vancomycin Hydrochloride Solutions at Various PH Values as Determined by High-Performance Liquid Chromatography

A stability-indicating HPLC assay method for the quantitation of vancomycin HCl has been developed. The developed method was used to study the effect of pH, phosphate buffer concentration and ionic strength on the stability of vancomycin HCl. The pH-rate profile curve showed 3 regions, acid-catalyzed, general base-catalyzed and the pH-independent region. The region of maximum stability was between pH 3.0 to about 5.7. On increasing the buffer concentration, phosphate ions hastened the decomposition of vancomycin HCl, the effect being more pronounced at pH 6.85 than at pH 4.43. In the pH region of 1-3, the decomposition was catalyzed by H⁺ and between pH 3-7 by HPO₄^2-, H₂PO₄^- and OH^-. An increase in the ionic strength decreased the rate of decomposition.     

高纯铝阳极氧化膜的电化学行为研究

为考察氧化膜状态对其电化学性能的影响,采用电化学方法研究了高纯铝阳极氧化膜在磷酸二氢钠缓冲溶液中的电化学性能.测试结果表明,高纯铝氧化膜的阴极极化过程分为3个阶段:1)电流密度变化较小而电位迅速负移,系克服氧化膜阻挡层电阻阶段;2)电流密度迅速增大而电位变化较小的析氢阶段;3)电流密度变化较小而电位迅速负移的氢离子扩散...     

The Influence of Potassium Clavulanate on the Rate of Amoxicillin Sodium Degradation in Phosphate and Acetate Buffers in the Liquid State

The stability of aqueous admixtures of amoxicillin sodium and potassium clavulanate was studied in the liquid state at selected pH values. Potassium clavulanate was found to catalyze the rate of degradation of amoxicillin sodium under the conditions of this study. In phosphate buffer (at pH 7.0) both amoxicillin sodium and potassium clavulanate showed first-order degradation when stored separately. However, when combined the rate of amoxicillin degradation increased and t₉₀ values for amoxicillin decreased from 69.6 min for amoxicillin alone to 10.8 min for amoxicillin in the combination at 55°C. A kinetic model was developed that explained the catalytic behavior of potassium clavulanate and phosphate buffer. In acetate buffer the rate of degradation of amoxicillin sodium followed first-order kinetics, but the catalytic effect of clavulanate caused curvature in the rate plots at higher temperatures and clavulanate concentrations. This catalytic effect was less than that occurred in phosphate buffer (where the t₉₀ value of amoxicillin decreased from 137.3 min for amoxicillin alone to 52.5 min for amoxicillin in combination at 55°C). First-order bi-exponential decay occurred with amoxicillin degradation, which explained this change in rate.     

Degradation of tobramycin in aqueous solution

The kinetics of degradation of tobramycin (Ne-De-Ka) in aqueous solution was studied as a function of pH. Tobramycin hydrolyzes in acidic solution to yield kanosamine (Ka-OH) and nebramine (Ne-De-OH) with a pseudo first-order rate constant of 2.7 × 10^-6 s^-1 in 1 N HCl at 80°C. The activation energy for the acid catalyzed hydrolysis is 32 kcal mol^-1. In basic solution, the hydrolysis products are deoxystreptamine (De-OH), nebramine (Ne-De-OH) and deoxystreptamine-kanosaminide (HO-De-Ka). The pseudo first-order rate constant for the hydrolysis in 1 N KOH is 1 × 10^-8 s^-1 at 80°C. The activation energy for the base catalyzed hydrolysis is 15 kcal mol^-1. Tobramycin is very stable towards hydrolysis at neutral pH; however, it rapidly oxidizes giving several products including De-OH, Ne-De-OH, and HO-De-Ka. In pH 7 phosphate buffer (0.01 M), the t₉₀ value is 70 hr at 80°C.     

Determination of the ionization state of 11-thioundecyl-1-phosphonic acid in self-assembled monolayers by chemical force microscopy

The present article describes the preparation and preliminary characterization of a novel phosphate-functionalized self-assembled monolayer (SAM) and the determination of the surface ionization states of the phosphate headgroup in aqueous solutions by chemical force microscopy (CFM). The phosphate headgroup used was PO(OH)2, a diprotic acid. The adhesion force between an AFM probe and a flat substrate, both of which were chemically modified with the same phosphate SAM, was also measured as a function of pH and ionic strength. At low ionic strength (10(-4) M), two peaks were observed in the force titration curve (adhesion force versus pH) at pH 4.5 and 8.4. The two peaks are positioned 2.4 and 1.2 pH units higher, respectively, than the acid dissociation constants obtained for the phosphate group free in aqueous solution. At high ionic strength (10(-1) M), the adhesion forces were reduced by 1 order of magnitude and the peaks were replaced by shoulders similar to those previously reported for acid force titrations. On the basis of JKR theory, the surface pKa values of the phosphate group in high ionic strength solutions were found to be 4.5 and 7.7, respectively. However, in light of the effects of ionic strength on the force titration curves, we discuss the applicability of JKR theory to nanoscopic measurements of adhesion force and surface pKa.     

Stabilisation of heavy metal containing dusts by reaction with phosphoric acid: study of the reactivity of fly ash

Water-washed fly ash was reacted with phosphoric acid in order to transform available heavy metals into insoluble metal phosphate compounds. The temperature, pH and concentration of free phosphate were monitored during the first 80 min of reaction. Phosphoric acid reacted rapidly with second order kinetics and an apparent rate constant of 0.015 l/(mol s m2). Analysis of the evolution of the concentrations of other major elements of fly ash shows that the reaction follows a dissolution-precipitation type mechanism. The solubility of trace heavy metals tends to increase at low pH values. Various heat and mass transfer coefficients are derived and help understand the phosphate stabilisation procedure and design industrial reactors for this purpose. Calcium phosphates are formed which can trap heavy metals in a stable apatite mineral structure.     

Growth imperfections in NH4H2PO4-KH2PO4 mixed crystals by X-ray topographic analysis

X-ray topographs of ammonium dihydrogen phosphate (ADP)—potassium dihydrogen phosphate (KDP) mixed crystals with various compositions were taken by the Lang transmission method. Growth imperfections such as growth bands, misorientations, lattice deformations at the boundary between two growth sectors and inclusions etc., present in the crystals were analysed. Dislocations were identified by using the g.b criterion.     

Sponge-like nickel phosphide–carbon nanotube hybrid electrodes for efficient hydrogen evolution over a wide pH range

Cost-effective hydrogen production via electrolysis of water requires efficient and durable earth-abundant catalysts for the hydrogen evolution reaction (HER) over a wide pH range.Herein,we report sponge-like nickel phosphidecarbon nanotube (NixP/CNT) hybrid electrodes that were prepared by facile cyclic voltammetric deposition of amorphous NixP catalysts onto the threedimensional (3D) porous CNT support.These compounds exhibit superior catalytic activity for sustained hydrogen evolution in acidic,neutral,and basic media.In particular,the NixP/CNT electrodes generate cathodic currents of 10 and 100 mA.cm-2 at overpotentials of 105 and 226 mV,respectively,in a 1 M phosphate buffer solution (pH =6.5) with a Tafel slope of 100 mV·dec-1;the currents were stable for over 110 h without obvious decay.Our results suggest that the 3D porous CNT electrode supports could serve as a general platform for earth-abundant HER catalysts for the development of highly efficient electrodes for hydrogen production.