EFFECTS OF ESCULETIN AS A LIPOXYGENASE INHIBITOR IN SOYBEAN EXTRACTS

This study was designed to determine the ability of esculetin to inhibit lipoxygenase activity in soybean. Lipoxygenase was extracted from ground soybean using 0.1 M Tris-HCl buffer (pH 8.6) and centrifuged at 3000 g for 15 min at 4C. The extract was mixed with different volumes of 0.05M esculetin solutions to provide 1.8 × 10⁻⁴M, 3.3 × 10⁻⁴M, 4.6 × 10⁻⁴M and 5.7 × 10⁻⁴M to final assay systems, respectively. Lipoxygenase activity decreased as the concentration of esculetin solution mixed in soybean extract increased. Mixing of 1.8 × 10⁻⁴M esculetin solution to soybean extract showed the least inhibition effect and was significantly different from that of 3.3 × 10⁻⁴M, 4.6 × 10⁻⁴M and 5.7 × 10⁻⁴M which inhibited LOX (lipoxygenase)'s activity of soybean extracts. In the study comparing the inhibition abilities for lipoxygenase activity among selected antioxidants, esculetin was significantly better than BHA (butylated hydroxyanisole) and α-tocopherol.     

Pacific Cod Muscle 5'-Nucleotidase

SUMMARY: A 5'-nucleotidase, widely distributed in teleost fish muscles, was purified about 20-fold from Pacific cod (Gadus macrocephalus) by chromatography of a dialyzed aqueous extract of the muscle on DEAE-cellulose. The enzyme was unstable and lost 85% of its activity in 1 hr at 37°C 53% in 10 min at 42°C and 40% in 1 hr at 30°C. It was stable for 6 days at 0°C, could be dialyzed for up to 3 days at 0°C against 1 mM tris buffer pH 7.5 and quickly frozen and thawed without loss of activity. However, it was inactivated rapidly when held at −30°C. Brief exposure to pH 4.0 or 5.0 effected marked destruction. Attempts at further purification by means of chromatography on hydroxylapatite, adsorption using alumina Cγ and starch gel electrophoresis failed due to instability.
The enzyme was strongly inhibited by EDTA, pyrophosphate, KF and ZnCl₂ (1-10 mM); less markedly inhibited by GSH, 2-mercaptoethanol, carbonate and CaCl₂ (10 to 100 mM). It was strongly activated by Mn⁺⁺ and weakly activated by Mg⁺⁺. The optimum pH was 7.6, and the Km was 5 × 10⁻⁴M with UMP and 8 ⁻⁴M with IMP. It hydrolyzed, in order of effectiveness, LJMP, IMP, CMP, d-AMP, GMP, d-IMP, d-GMP, d-UMP and AMP, but not p-nitro phenylphosphate, sugar phosphates or a number of other compounds including 2',3'-nucleotides.     

Singlet Oxygen Oxidation Rates of ∝-, γ-, and δ-Tocopherols

ABSTRACT:  The reaction rate constants of 5 × 10⁻⁴ M, 10 × 10⁻⁴ M, and 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols with singlet oxygen in methylene chloride containing 1 × 10⁻⁵ M chlorophyll under light at 25 °C for 60 min were studied. The oxidation of tocopherols determined by a spectrophotometric method showed that the losses of 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols after 60 min under light were 21%, 16%, and 9%, respectively. The degradation of α-, γ-, and δ-tocopherols was undetectable in the absence of chlorophyll under light or in the presence of chlorophyll in dark. The losses of tocopherols under light were mainly due to singlet oxygen oxidation. The degradation rates of 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols were 6.6 ×10⁻⁶ M/min, 5.0 × 10⁻⁶ M/min, and 2.9 × 10⁻⁶ M/min, respectively. The reaction rates between α-, γ-, or δ-tocopherol and singlet oxygen were 4.1 ×10⁶/M s, 3.3 × 10⁶/M s, and 1.4 × 10⁶/M s, respectively. The singlet oxygen oxidation rate of δ-tocopherol was significantly lower than α- or γ-tocopherol at α= 0.05. As the electron density in the chromanol ring of tocopherol increased, the singlet oxygen oxidation was increased.     

EFFECT OF SEVERAL EXPERIMENTAL PARAMETERS ON COMBINATION OF RED KIDNEY BEAN (PHASEOLUS VULGARIS) α-AMYLASE INHIBITOR WITH PORCINE PANCREATIC α-AMYLASE

Purified red kidney bean (Phaseolus vulgaris) amylase inhibitor forms a 1:1 stoichiometric complex with porcine pancreatic α-amylase leading to complete loss of enzyme activity on starch. Rate of complex formation is pH dependent and is maximal at pH 5. The rate constants for complex formation, as measured by loss of amylase activity, were 2.85 × 10⁴ M^-1 sec^-1 at pH 6.9 (ionic strength of 0.918) and 2.55 × 10⁵ M^-1 sec^-1 at pH 5 at 30°C. At pH 6.9, rate of complex formation was 4.8 times faster at 0.918 ionic strength as compared with the rate at 0.138 ionic strength. At 30°C, pH 6.9 and ionic strength of 0.168 the dissociation constant of the enzyme-inhibitor complex was determined to be 3.5 × 10^-11 M. The rate constant for dissociation of the complex was calculated to be 8.7 × 10^-8 sec^-1 under the same conditions. The rate constant for complex formation, at ionic strength of 0.168, was 1.1 × 10⁴ M^-1 sec^-1 at 37⁰ and 9.77 × 10² M^-1 sec^-1 at 25.7°C. The calculated activation energy for complex formation is 39.5 kcal/mole suggesting a rate-controlling conformational change. Oxidation of the carbohydrate moiety of the glycoprotein inhibitor caused complete loss of activity. Maltose, a competitive inhibitor of α-amylase, bound as readily to the enzyme-inhibitor complex as to free α-amylase. Trypsinized α-amylase, although still able to bind to Sephadex, did not bind inhibitor. The experiments with maltose and trypsinized amylase suggest the inhibitor may not bind at the active site of α-amylase.     

Isolation and characterization of polygalacturonase and invertase from a yeast Endomycopsis fibuligera (3812) bioconversion of date wastes

Properties of polygalacturonase [E.C. 3.2.1.15] (E₁) and invertase (E.C. 3.2.1.26] (E₂) isolated and purified in powdered form from Endomycopsis fibuligera grown on date wastes were measured.
The molecular weights were estimated to be about 32000 and 69500, and they were highly specific for hydrolysing (1–4) and (1–2) glycosidic links respectively. the K_m value of E₁ was 7.1 × 10^-2M for soluble-pectic acid and that for E₂ was 6.7 × 10^-2M for sucrose at pH4.5, and optimal pHs were 3.5 for E₁ and 4.0 for E₂. the purified enzymes are stable on storage for more than one year at - 20°C.     

Quenching Mechanisms and Kinetics of α-Tocopherol and β-Carotene on the Photosensitizing Effect of Synthetic Food Colorant FD&C Red No. 3

ABSTRACT: Effects of FD&C Red No. 40, Red No. 3, Yellow No. 5, Yellow No. 6, Green No. 3, Blue No. 1 and Blue No. 2 on 0.03M soybean oil oxidation in acetone at 25 °C under light were studied by measuring headspace oxygen depletion. As Red No. 3 increased from 0 to 5, 20, 100 and 200 ppm, the headspace oxygen was reduced by 2 to 70, 73, 77 and 77%, respectively, for 4 h. Only Red No. 3 acted as a photosensitizer to produce singlet oxygen in the oil. The quenching rates of α-tocopherol and β-carotene for the singlet oxygen by Red No.3 were 4.1 × 10⁷ M⁻¹s⁻¹ and 7.3 × 10⁹ M⁻¹s⁻¹, respectively. When β-carotene was below 1.86 × 10⁻⁶ M, β-carotene quenched singlet oxygen, but it quenched both singlet oxygen and Red No. 3 at or above 3.72 × 10⁻⁶ M. However, α-tocopherol quenched singlet oxygen only.     

CHARACTERIZATION OF PHYTASE OF BEANS (PHASEOLUS VULGARIS)

Phytase from California Small White beans was extracted, concentrated and enriched by heat treatment and ammonium sulfate fractionation. Crude enzyme fractions tenaciously retained endogenous phytate which could be removed by autolysis at 45°C and pH 5.2. Characterization of the phytase preparation showed: (1) increase in activity with increasing temperature from 37 to 60°C, (2) an activation energy of approximately 9,200 cal/mole for the hydrolysis of inositol hexaphosphate; (3) pH optimum of 5.2; (4) a K_m value of 2.22 × 10^-4M; (5) apparently partial inhibition at high substrate concentrations; (6) fully competitive inhibition by one of the reaction products, inorganic phosphate, with K_i=3.1 × 10⁴M. These properties and kinetic constants are comparable to those reported for phytase preparations from similar sources and they adequately account for in situ autolytic loss of phytic acid from these beans.     

Effect of technological parameters on the characteristics of kasseri cheese made from raw or pasteurized ewes' milk

Two groups of kasseri cheese (pasta filata type) were manufactured from raw or pasteurized ewes' milk, without starter cultures. Cheeses of each group were divided into two subgroups: the first was ripened and stored at 4°C and packaged in plastic film; the second ripened and stored at 15°C and coated with paraffin wax. Milk pasteurization and technological parameters had a significant effect on the pH ( P  < 0.05), while only technological parameters had an effect on the total solids content. At day 120, the range of mean cfu/g counts for the mesophilic aerobic flora was 9.5 × 10⁷−1.4 × 10⁸; for the thermophilic streptococci, the range was 2.6 × 10⁷−7.6 × 10⁷; and for the thermophilic bacilli, 9.8 × 10⁶−1.7 × 10⁷. Changes in the N fractions became significant after 30 days of ripening. For mature 120-day-old cheeses, the percentage of total N soluble at pH 4.6 was 22.7%–22.9% in raw milk cheeses and 19.0%–21.7% in pasteurized milk cheeses. The percentage of total N soluble at 12% TCA was 10.1%–12.2% in raw milk cheeses and 7.3%–11.5% in pasteurized milk cheeses; the percentages of total N soluble at 5% PTA were 3.1%–4.0% and 2.6%–3.6%, respectively. The residual α_s-casein percentages at day 120 ranged between 63% and 78% of the respective area at day 1; the residual β-casein ranged between 67% and 75%. There were some characteristic differences in the reverse phase-HPLC peptide profiles of the four cheeses. In general, the effect of the different ripening conditions was more pronounced in cheeses made from pasteurized milk.     

Natural Antioxidants as a Component of an Egg Albumen Film in the Reduction of Lipid Oxidation in Cooked and Uncooked Poultry

ABSTRACT: Egg albumen coatings with natural antioxidants fenugreek, rosemary, and vitamin E were evaluated for their antioxidant activity in diced raw and diced cooked poultry breast meat. Coatings were applied using a vacuum tumbling and a boiling method. Coated samples and uncoated controls were evaluated for their malondialdehyde and reactive substances content as a measure of lipid oxidation via the thiobarbituric acid test. Cooked egg albumen-coated samples showed reactive compound changes of 1.56 × 10⁻⁷M for 4 d of refrigeration, compared to 1.59 × 10⁻⁶ M change in the cooked control. Raw control changed by 1.97 × 10⁻⁶M versus 1.67 × 10⁻⁶ M for raw rosemary coated samples. In raw and cooked studies EAS coating showed most effective against lipid oxidation.     

BACTERIAL FORMATION OF HISTAMINE IN JACK MACKEREL (TRACHURUS SYMMETRICUS)

Peak histamine concentrations of 0.023, 0.031 and 0.027 g histamine/100 g muscle and maximal bacteria concentrations of 1.75, 1.59 and 0.423 g dry cells/100 g muscle were observed in muscles of jack mackerel stored at 25, 15 and 5C, respectively. Incubated fish homogenates suggest rate and transport limitations in histamine formation in muscle. The Mulchandani model predicted bacterial growth in muscle. The Luedeking and Piret expression fitted histamine formation in muscle; α values were 3.0 × 10⁻³, 1.23 × 10⁻² and 4.17 × 10⁻² g histamine/g dry cells, while β-values were 4.5 × 104, 8.0 × 10⁻⁵ and 0 g histamine/g dry cells × h at 25, 15, and 5C, respectively. The model predicts that jack mackerel could be stored from 4.5 to 5.5 days in ice, from 1 to 2 days at 15C and from 17 h to 2 days at 25C before fishmeal quality might be affected.     

Development of a Meat-Like Process Flavoring from Soybean-Based Enzyme-Hydrolyzed Vegetable Protein (E-HVP)

ABSTRACT: Response surface methodology (RSM) was used to develop a meat-like process flavoring agent from enzyme-hydrolyzed vegetable protein (E-HVP). Five factors were evaluated: pH (3.6 to 8.4), temperature (51 to 99 °C), heating period (0.3 to 2.7 h), amount of ribose (0 to 1 × KHmol) and amount of cysteine (0 to 1 × 10 ³mol). Sensory analysis limited to aroma in terms of overall liking and intensity of specific aroma attributes was investigated. The aroma attributes measured included bean-like, potato-like, Brussels sprouts-like, molasses-like, chicken-like, beef-like, egg-like, roasted and apple sauce-like). Based on the fitted surfaces and consumer test data (overall liking), the optimum reaction conditions for production of a meat-like flavoring were pH 6,99 °C reaction temperature, 1.5 h heating time, 5 × 10⁻⁴ mol of ribose and 5 × 10⁻⁴ mol of cysteine.     

Purification and Characterization of an α-L-Rhamnosidase from Aspergillus terreus of Interest in Winemaking

ABSTRACT: An enzyme with α-L-rhamnosidase activity was purified to homogeneity from a culture filtrate of Aspergillus terreus after growth in a medium containing L-rhamnose as the sole carbon source. The biosynthesis of this enzyme was repressed by glucose. The enzyme had a molecular mass of 96 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of 4.6 as determined by analytical isoelectric focusing. The pH and temperature optima for the enzyme were found to be 4.0 and 44 °C, respectively. Using p-nitrophenyl-α-L-rhamnopyranoside as a substrate, the enzyme exhibited Michaelis-Menten kinetics with K_M and V_max values of 0.17 mM and 84 U/mg, respectively. The enzyme was inhibited competitively by L-rhamnose (K₁ 2.5 mM). Divalent cations such as Ca²⁺ Mg²⁺ Zn²⁺ and Co²⁺ stimulated the a-L-rhamnosidase activity, whereas this was inhibited by Hg2+ and Cd2+. Ethanol (12% v/v) and glucose (21% w/v) decreased enzyme activity by approximately 20%, while this was not affected by SO₂.     

Whey Protein Film Composition Effects on Potassium Sorbate and Natamycin Diffusion

ABSTRACT: To assess the ability of whey protein films to act as antimicrobial carriers, the effect of film composition on preservative diffusion was investigated. Preservative diffusion coefficients were measured at 24°C in whey protein isolate (WPI) films with different WPI-glycerol plasticizer ratios (1:1 to 15:1), beeswax (BW) content, 0% to 40% w/w dry solids, and preservative addition of 0.3% (w/w) natamycin or 1.6% (w/w dry solids) potassium sorbate. Diffusion coefficients for potassium sorbate and natamycin were in the ranges 1.09 × 10⁻¹¹ to 13.0 × 10⁻¹¹ m²/s and 6.16 × 10⁻¹⁴ to 37.8 × 10⁻¹⁴ m²/s, respectively, and significantly decreased as the WPI-glycerol ratio increased. No significant difference in sorbate diffusion was seen with the addition of BW.     

Inverse Method to Estimate Kinetic Degradation Parameters of Grape Anthocyanins in Wheat Flour Under Simultaneously Changing Temperature and Moisture

ABSTRACT:  Thermal and moisture effects on grape anthocyanin degradation were investigated using solid media to simulate processing at temperatures above 100 °C. Grape pomace (anthocyanin source) mixed with wheat pastry flour (1: 3, w/w dry basis) was used in both isothermal and nonisothermal experiments by heating the same mixture at 43% (db) initial moisture in steel cells in an oil bath at 80, 105, and 145 °C. To determine the effect of moisture on anthocyanin degradation, the grape pomace–wheat flour mixture was heated isothermally at 80 °C at constant moisture contents of 10%, 20%, and 43% (db). Anthocyanin degradation followed a pseudo first-order reaction with moisture. Anthocyanins degraded more rapidly with increasing temperature and moisture. The effects of temperature and moisture on the rate constant were modeled according to the Arrhenius and an exponential relationship, respectively. The nonisothermal reaction rate constant and activation energy (mean ± standard error) were k _{80 °C, 43% (db) moisture} = 2.81 × 10⁻⁴± 1.1 × 10⁻⁶ s⁻¹ and Δ E  = 75273 ± 197 J/g mol, respectively. The moisture parameter for the exponential model was 4.28 (dry basis moisture content)⁻¹. One possible application of this study is as a tool to predict the loss of anthocyanins in nutraceutical products containing grape pomace. For example, if the process temperature history and moisture history in an extruded snack fortified with grape pomace is known, the percentage anthocyanin loss can be predicted.     

Changes in the Quality of Fresh-cut Jicama in Relation to Storage Temperatures and Controlled Atmospheres

ABSTRACT: Intact jicama (Pachyrhizus erosus) roots are chilling sensitive, but quality of fresh-cut pieces (1.8 × 4.5 cm cylinders) was best maintained at low storage temperatures (0 to 5 °C). Respiration rates of different piece sizes were similar, and averaged 2, 7 and 10 μL CO₂-g ⁻¹h⁻¹ at 0 °C, 5 °C and 10 °C, respectively. Storage in air at 5 °C to 10 °C resulted in surface browning and was associated with increases in phenolics and phenylalanine ammonia lyase and polyphenol oxidase activities. High CO₂ atmospheres (5 to 10%) at 5 °C were very effective in retarding microbial growth and discoloration. The source of jicama root notably affected the quality and shelf-life of the fresh-cut pieces. Fresh-cut pieces from stored roots (2 wk at 19 to 22 °C) had lower visual quality and crispness during subsequent storage than did pieces from recently harvested roots.     

The air drying behaviour of fresh and osmotically dehydrated banana slices

Ripe banana, cut to 10mm thick slabs were osmotically treated in sugar solutions of 35, 50 and 65° Brix for 36h. The initial moisture content fell from a value of 3.13kg H₂O DM to 2.19, 1.63 and 1.16kg H₂O kg⁻¹ for treatment in the three solutions, respectively. These slabs, with Total Soluble Solids (TSS) contents of 26, 34 and 39° Brix, respectively, as well as freshly cut but untreated slabs (15° Brix) were air dried in a cabinet type tray drier to near equilibrium conditions at fixed temperatures from 40 to 80°C and at a constant air speed of 0.62m s⁻¹. Drying was found to occur in the falling rate period only for both banana types and two drying constants K₁ and K₂ were established for a first and second falling rate period of drying. Increasing the drying air temperature significantly enhanced the drying rate and the K-values, except at 80°C when the rates fell, possibly because of case hardening of the slabs. Reducing the slab thickness also improved the drying rate, but increasing the air speed to 1.03m s⁻¹ did not have any profound effect. As the sugar content of the banana slabs increased through the osmotic treatment, drying rates fell. Calculated apparent moisture diffusivities at 60°C ranged from 34.8× 10⁻¹⁰ m² s⁻¹ (fresh slab) to 8.8×10⁻¹⁰ m² s⁻¹ for dried (39° Brix) slabs. The moisture diffusivity was significantly lowered as the moisture content dropped in drying and with increased levels of sugar. Previously osmosed and then air dried banana slabs showed appealing colour and texture compared to the fresh banana.     

Alginate- and gellan-based edible films for probiotic coatings on fresh-cut fruits

ABSTRACT:  Alginate- (2% w/v) or gellan-based (0.5%) edible films, containing glycerol (0.6% to 2.0%), N-acetylcysteine (1%), and/or ascorbic acid (1%) and citric acid (1%), were formulated and used to coat fresh-cut apple and papaya cylinders. Water vapor permeability (WVP) was significantly higher ( P < 0.05) in alginate films (0.30 to 0.31 × 10⁻⁹g m/Pa s m²) than in the gellan ones (0.26 to 0.27 × 10⁻⁹g m/Pa s m²). Addition of 0.025% (w/v) sunflower oil decreased WVP of gellan films (0.20 to 0.22 × 10⁻⁹ g m/Pa s m²). Water solubility of gellan and alginate films at 25 °C (0.47 to 0.59 and 0.74 to 0.79, respectively) and their swelling ratios (2.3 to 2.6 and 1.6 to 2.0, respectively) indicate their potential for coating high moisture fresh-cut fruits. Fresh-cut apple and papaya cylinders were successfully coated with 2% (w/v) alginate or gellan film-forming solutions containing viable bifidobacteria. WVP in alginate (6.31 and 5.52 × 10⁻⁹g m/Pa s m²) or gellan (3.65 and 4.89 × 10⁻⁹ g m/Pa s m²) probiotic coatings of papaya and apple, respectively, were higher than in the corresponding cast films. The gellan coatings and films exhibited better water vapor properties in comparison with the alginate coatings. Values > 10⁶ CFU/g B. lactis Bb -12 were maintained for 10 d during refrigerated storage of fresh-cut fruits, demonstrating the feasibility of alginate- and gellan-based edible coatings to carry and support viable probiotics on fresh-cut fruit.     

Thermal Kinetic Parameters of Thiamin in Wheat Flour at Temperatures Higher than 100°C

ABSTRACT: Kinetic parameters for thiamin degradation were obtained using 2 high-temperature heating methods: (1) atmospheric pressure (AP) with moisture correction and (2) controlled pressure (CP). At AP conditions, 33.3% dry basis (db) moisture wheat flour with 0.35% (db) thiamin was heated in thin steel cells isothermally at 145,160, and 172°C. To obtain the moisture correction factor, a constant-moisture study was conducted at 80°C using 6 moisture contents (6.1% to 36.9%). At CP conditions, flour at 19%, 28.2%, and 33.3% (db) moisture in double-seamed cans was heated in a CP steam retort at 129°C. For the AP method, the corrected activation energy for 33.3% moisture content was 129.5 kJ/g-mol and reaction rate at 80°C was 3.48×10⁻⁴ min⁻¹. Using the CP method, the activation energy and reaction rate were 121.0 kJ/g-mol and 9.69×10⁻⁵ min⁻¹, respectively. Results obtained from 2 methods were not statistically different. These results illustrated that the correction method could be used as an alternative for researchers without access to controlled pressure equipment and transient heat transfer software.     

Microbial flora of technological interest in raw ovine milk during 6°C storage

Changes in the microbial flora of ovine milk of medium hygienic quality (initial mean total plate count 3.3  ×  10⁵ cfu/mL) were studied throughout refrigerated storage at 6°C. Total plate counts after 48 h of refrigerated storage (mean count 4.6  ×  10⁵ cfu/mL) were under the current standards in the European Union (EU) for raw ovine milk to be used for cheesemaking after heat treatment. However, after 96 h, mean total plate counts (1.6  ×  10⁷ cfu/mL) were above the standards. Lactococci were found at levels higher than Pseudomonas spp. in milk freshly drawn (54.5% and 3.5% of total plate count, respectively) and after 96 h at 6°C (47.9% and 33.9% of total plate count, respectively). Lactobacilli, enterococci, coliforms and thermodurics were found at values lower than lactococci and Pseudomonas spp., before and after refrigerated storage ( ≤  0.08% of the total plate count). A significant growth ( P  < 0.001) was detected for mesophiles, Pseudomonas spp. and lactococci after 96 h of refrigeration at 6°C; however, thermodurics, coliforms, lactobacilli and enterococci, showed no significant increase ( P  > 0.05). Presumptive Escherichia coli (β-glucuronidase-positive) underwent a decrease throughout storage at 6°C.     

Osmotic dehydration kinetics of carrot cubes in sodium chloride solution

Osmotic dehydration kinetics of carrot cubes in sodium chloride solution having concentrations 5%, 10% and 15% (w/v), solution temperature 35, 45 and 55 °C, sample to solution ratio (STSR) 1:4, 1:5 and 1:6 were studied up to 240 min duration. During the experimentation, effect of solution temperature and process duration was significant and that of solution concentration and STSR were non-significant on water loss. Among the different models applied (Penetration model, Magee Model, and Azuara model), Azuara model best fitted to the experimental data for water loss and solute gain during osmotic dehydration. Effective diffusivities of water and solute were calculated by using the analytical solution of Fick's unsteady state law of diffusion by iterative technique with a computer program. For the above conditions, the effective diffusivity of water was found to be in the range between 2.6323 × 10⁻⁹ and 6.2397 × 10⁻⁹ m²/s and that of solute between 3.1522 × 10⁻⁹ and 4.6400 × 10⁻⁹ m²/s.