AbstractThe solubilities of l-glutamine in mixed solvents (ethanol + water, acetone + water) were determined at temperatures between 278 K and 313 K. The solubilities of l-glutamine in mixed solvents (water + ethanol, water + acetone) are a function of temperature, increasing with an increase in temperature. Pure water has a high dissolving power. The experimental data were correlated with the Apelblat model. The calculated values of Apelblat model were found to show a fine representation of the experimental data. Then the dissolution enthalpy and entropy of l-glutamine were predicted from the solubility data using van’t Hoff equation. The dissolution enthalpy and entropy of l-glutamine in water predicted from the solubility data is less than that in mixed solvents.Highlights► The solubilities of l-glutamine in mixed solvents (ethanol + water, acetone + water) were measured. ► It increases with an increase in temperature. ► Pure water has a high dissolving power. ► The calculated solubility sets a good coherence with the experimental values. ► The dissolution enthalpy and entropy were predicted using van’t Hoff equation.KeywordsSolubility; l-Glutamine; Dissolution enthalpy; Dissolution entropy1. Introductionl-Glutamine (l-Gln, Fig. 1), a derivative of l-glutamate (l-Glu), is considered as a conditionally essential amino acid and a promising new drug [1]. It is the most abundant amino acid in blood, and is a carrier or nitrogen transport. l-Gln plays an important role in regulating the synthesis of protein, nucleic acid and lycoprotein. Medical studies have indicated that a shortage of l-Gln could cause many diseases [2]. l-Gln is mainly produced by microbiological fermentation. The manufacturing process of an amino acid by fermentation comprises fermentation, crude isolation and purification processes. Final purification is performed to ensure the required quality for the intended use. The final product is obtained as a crystalline powder. In the final purification step, l-Gln is often refined through crystallization from solution. But the solubility of l-Gln in water is barely affected by temperature [3]. In addition to this, the higher temperature favors l-Gln degradation [4]. Consequently, cooling crystallization for which the precondition is to heating the system is not applicable for harvesting l-Gln.Chemical structure of l-glutamine.Fig. 1. Chemical structure of l-glutamine.Figure optionsl-Gln is stable around the isoelectric point (pH 5.65), but if the pH shifts from the isoelectric point to either acid or alkaline conditions, l-Gln is easily hydrolyzed to l-glutamic acid and ammonia. To find a proper solvent with the view of drowning-out crystallization and avoiding degradation, an experimental work devoted to collect information on the solubility of l-Gln in the ternary system ethanol + water, acetone + water was carried out.Innumerable techniques, of almost infinite variety, have been proposed at one time or another for the measurement of the solubility of solids in liquids. No single method can be identified, however, as being generally applicable to all possible types of system. The choice of the most appropriate method for a given case has to be made in the light of the system properties, the availability of apparatus and analytical techniques, the skill and experience of the operators, the precision required, and so on [5] and [6]. Due to the fact that l-glutamic acid, the degradation product of l-Gln, has almost equal molar weight 147.15, with l-Gln 146.17, l-Gln degradation would not affect the saturated solution concentration measurement by weighing method. Therefore, gravimetric method is feasible to measure the solubility of l-Gn in the low temperature range.2. Experimental2.1. Materialsl-Gln was purchased from Sigma–Aldrich and used without any further purification. The mass fraction purity is >0.99. The ethanol and acetone used for experiments were of analytical reagent grade. Their mass fraction purities were greater than 0.99.2.2. ProcedureThe solubility measurement by gravimetric method includes two steps, first the preparation of saturated solution of l-Gln in mixture solvent at the given temperature and then the concentration determination of saturated solution.2.2.1. The preparation of saturated solutionThe traditional preparation process by isothermal method is as follows [5], [6] and [7]. The cylindrical double-jacketed glass vessel was controlled to a constant temperature with continuous stirring, in which the temperature was controlled to be constant with a precision of ±0.05 K through a thermostatic bath (Type 501 A, China). Then, excess solute was added to the solvent. After the temperature of the water bath was kept constant for at least 4 h, the stirring was stopped, and the solution was kept still for 3 h. A portion of the upper clear solution was filtered with the membrane (0.45 μm).In terms of this method a long dissolution time (4 h) has an increased risk of l-Gn degradation. Therefore, an experiment to find a proper dissolution time is carried out. Excess solute was added to the solvent, the concentration of suspension was measured by weighing method (described later) at different dissolve times. Fig. 2 gives the concentration variation of suspension along with dissolution time. The suspension can reach the dissolution phase equilibrium in short term, <2 h. So in this work the dissolution time is taken as 2 h.l-Glutamine suspension concentration at different dissolution times.Fig. 2. l-Glutamine suspension concentration at different dissolution times.Figure options2.2.2. The concentration determination of saturated solutionA portion of the upper clear solution prepared by the above method was filtered with the membrane (0.45 μm) and preserved in a weighted double dish which had been completely dried. The double dish was quickly weighed to determine the mass of the sample. The solvent in the double dish was completely evaporated after 10 h with the oven at 60 °C; the double dish was reweighed to determine the mass of the residue solid and the evaporated solvent. All of the masses were measured using an electronic balance with an uncertainty of 0.0001 g.During the drying process at 60 °C l-Gln degradation is inevitable. To this end, an experiment was conducted to test the influence of this drying process on the measured concentration of l-Gn in solution. A series of solutions with different concentrations of l-Gn were dried by the above process. The calculated concentration based on the mass of the residue solid and the evaporated solvent and the actual solution concentration were compared in Fig. 3. The results showed this drying process in experiment does not affect the concentration calculation of l-Gn in solution.
The measured concentration of a series of different concentration solution.
Fig. 3.
The measured concentration of a series of different concentration solution.
Figure options
The mole fraction solubility (x1) of l-Gn in the solvent system was obtained as follows:
equation(1)
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where m1, m2, and m3 represent the masses of solute, water, and the other solvent in the mixed solvent system. M1, M2, and M3 are the molecular weights of solute, water, and the other solvent in the mixed solvent system. All of the experiments were repeated three times at each temperature. The relative uncertainty of the experimental solubility values is within 0.05.
3. Results and discussion
The results of l-Gn solubility in water and mixed solvents are listed in Table 1. Fig. 4 gives the plot of the solubility of l-Gn in these solvents at a temperature range of about 278–313 K. In all of the tables and figures, R, which represents the solvent molar ratio of the solvent mixture, can be calculated as follows:
equation(2)
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where Ni represents the other solvents except for water, such as ethanol and acetone. The solubilities are a function of temperature, increasing with an increase in temperature. In the mixed solvent systems, the solubility would increase with the increasing of R, which indicates that pure water has a high dissolving power. Among the two different mixed solvent systems, only if it has the same solvent molar ratio (R), the molar fraction solubility will decrease in the same degree.
Table 1.
Mole fraction solubility x1 of l-glutamine in different solvents as a function of temperature T along with differences from the calculated mole fraction xcalcd of Eq. (3) with parameters of Table 2 at pressure p = 0.1 MPa a (R is molar ratio, R = Nwater/Nacetone or ethanol).
T (K) 103x1 102(x1 − xcalcd)/x1 T (K) 103x1 102(x1 − xcalcd)/x1
Water Acetone + water, R = 10
278.15 2.54 −0.70 278.15 0.79 −4.78
283.15 3.05 −0.46 283.15 1.04 1.63
288.35 3.68 0.63 288.15 1.27 1.16
293.15 4.25 −0.85 293.15 1.57 2.10
298.15 5.14 2.01 298.15 1.85 −0.54
303.15 6.02 2.10 303.15 2.23 −0.34
308.15 6.62 −2.93 308.15 2.64 −1.40
313.15 7.95 1.11 312.95 3.18 0.77
Acetone + water, R = 20 Acetone + water, R = 40
277.95 1.46 0.01 277.95 1.82 −1.31
282.95 1.71 −1.21 282.95 2.23 0.25
287.95 2.07 0.59 287.85 2.70 1.36
292.65 2.42 0.23 292.75 3.18 0.71
297.55 2.90 1.77 297.65 3.70 −0.82
302.65 3.31 −2.46 302.85 4.36 −1.69
307.75 4.06 0.71 307.73 5.26 1.63
312.65 4.74 −0.19 312.65 6.00 −0.40
Ethanol + water, R = 10 Ethanol + water, R = 20
278.15 0.84 1.39 278.15 1.33 −0.35
283.15 1.05 −1.61 283.15 1.65 −0.94
288.15 1.31 1.14 288.15 2.06 0.48
293.15 1.62 −2.04 293.15 2.50 −0.28
298.15 2.00 1.27 298.15 3.10 2.25
303.15 2.44 0.36 303.15 3.59 −1.19
308.15 2.97 −0.52 308.15 4.28 −0.74
313.15 3.60 0.12 313.15 5.09 0.47
Ethanol + water, R = 40
278.15 1.82 0.26 298.15 3.79 −0.96
283.15 2.20 −0.63 303.15 4.55 0.18
288.15 2.66 −0.44 308.15 5.36 −0.11
293.15 3.26 1.55 313.15 6.30 0.05
a
Standard uncertainties u are u(T) = 0.05 K, ur(p) = 0.05, ur(x1) = 0.05.
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