Thermochimica Acta 558 (2013) 69

Contents lists available at SciVerse ScienceDirect

Thermochimica Acta
journal homepage: www.elsevier.com/locate/tca

Solubility, dissolution enthalpy and entropy of l-glutamine in mixed solvents of

ethanol + water and acetone + water
Qiushuo Yu a,b,c , Xiaoxun Ma a,b,c, , Long Xu a,b,c
a
School of Chemical Engineering, Northwest University, Taibai North Road 229, Xian, Shaanxi 710069, China
b
Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, China
c
Shaanxi Research Center of Engineering Technology for Clean Coal Conversion, China

a r t i c l e i n f o a b s t r a c t

Article history: The solubilities of l-glutamine in mixed solvents (ethanol + water, acetone + water) were determined at
Received 14 December 2012 temperatures between 278 K and 313 K. The solubilities of l-glutamine in mixed solvents (water + ethanol,
Received in revised form 2 February 2013 water + acetone) are a function of temperature, increasing with an increase in temperature. Pure water has
Accepted 5 February 2013
a high dissolving power. The experimental data were correlated with the Apelblat model. The calculated
Available online 19 February 2013
values of Apelblat model were found to show a ne representation of the experimental data. Then the
dissolution enthalpy and entropy of l-glutamine were predicted from the solubility data using vant Hoff
Keywords:
equation. The dissolution enthalpy and entropy of l-glutamine in water predicted from the solubility
Solubility
l-Glutamine
data is less than that in mixed solvents.
Dissolution enthalpy 2013 Elsevier B.V. All rights reserved.
Dissolution entropy

1. Introduction To nd a proper solvent with the view of drowning-out crystalliza-

l-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 regu-
lating 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 fermenta-
tion. The manufacturing process of an amino acid by fermentation
comprises fermentation, crude isolation and purication processes.
Final purication is performed to ensure the required quality for the
intended use. The nal product is obtained as a crystalline pow-
der. In the nal purication step, l-Gln is often rened 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.
l-Gln is stable around the isoelectric point (pH 5.65), but if the
pH shifts from the isoelectric point to either acid or alkaline condi-
tions, l-Gln is easily hydrolyzed to l-glutamic acid and ammonia.
tion 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 innite variety, have been
proposed at one time or another for the measurement of the sol-
ubility of solids in liquids. No single method can be identied,
however, as being generally applicable to all possible types of sys-
tem. The choice of the most appropriate method for a given case
has to be made in the light of the system properties, the availabil-
ity of apparatus and analytical techniques, the skill and experience
of the operators, the precision required, and so on [5,6]. 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. Experimental
2.1. Materials

l-Gln was purchased from SigmaAldrich and used without any

Corresponding author at: School of Chemical Engineering, Northwest University,
further purication. The mass fraction purity is >0.99. The ethanol
Taibai North Road 229, Xian, Shaanxi 710069, China. Tel.: +86 029 88307657;
fax: +86 029 88302223.
and acetone used for experiments were of analytical reagent grade.
E-mail addresses: yqiushuo@126.com (Q. Yu), maxym@nwu.edu.cn (X. Ma). Their mass fraction purities were greater than 0.99.

0040-6031/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.tca.2013.02.007
 Q. Yu et al. / Thermochimica Acta 558 (2013) 69 7

0.16

The measured concentration/(g/g solvent)

Experiment data
0.14
The line of y=x
0.12

0.10
Fig. 1. Chemical structure of l-glutamine.
0.08

2.2. Procedure 0.06

The solubility measurement by gravimetric method includes
two steps, rst the preparation of saturated solution of l-Gln in mix-
ture solvent at the given temperature and then the concentration
determination of saturated solution.
2.2.1. The preparation of saturated solution
The traditional preparation process by isothermal method is as
follows [57]. The cylindrical double-jacketed glass vessel was con-
trolled 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 ltered with the membrane (0.45 m).
In terms of this method a long dissolution time (4 h) has an
increased risk of l-Gn degradation. Therefore, an experiment to
nd a proper dissolution time is carried out. Excess solute was
added to the solvent, the concentration of suspension was mea-
sured 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.
2.2.2. The concentration determination of saturated solution
A portion of the upper clear solution prepared by the above
method was ltered with the membrane (0.45 m) and preserved
in a weighted double dish which had been completely dried. The
double dish was quickly weighed to determine the mass of the sam-
ple. The solvent in the double dish was completely evaporated after
10 h with the oven at 60 C; the double dish was reweighed to deter-
mine the mass of the residue solid and the evaporated solvent. All
0.04
0.02
0.00
-0.02
-0.02 0.00 0.02 0.04 0.06 0.08 0.10 0.12 0.14 0.16
The actual concentration / (g/g solvent)
Fig. 3. The measured concentration of a series of different concentration solution.
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
inuence 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 concen-
tration 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 mole fraction solubility (x1 ) of l-Gn in the solvent system
was obtained as follows:
m1 /M1
x1 = (1)
m1 /M1 + m2 /M2 + m3 /M3
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

0.04
listed in Table 1. Fig. 4 gives the plot of the solubility of l-Gn in
these solvents at a temperature range of about 278313 K. In all of
the tables and gures, R, which represents the solvent molar ratio
concentration / (g/g solvent)

of the solvent mixture, can be calculated as follows:

0.03
Nwater
R= (2)
Ni
0.02 where Ni represents the other solvents except for water, such as
ethanol and acetone. The solubilities are a function of temper-
ature, increasing with an increase in temperature. In the mixed
0.01 solvent systems, the solubility would increase with the increas-
ing of R, which indicates that pure water has a high dissolving
power. Among the two different mixed solvent systems, only if it
0.00
has the same solvent molar ratio (R), the molar fraction solubility
will decrease in the same degree.
The solubility can be correlated by a semi-empirical equation
0 1 2 3 4 5 (Apelblat model) as follows [8]:
Time / hour
B
ln x1 = A + + C ln(T /K) (3)
Fig. 2. l-Glutamine suspension concentration at different dissolution times. T /K
8 Q. Yu et al. / Thermochimica Acta 558 (2013) 69

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 MPaa (R is molar ratio, R = Nwater /Nacetone or ethanol ).

T (K) 103 x1 102 (x1 xcalcd )/x1 T (K) 103 x1 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.

where T is temperature and A, B and C are all empirical constants The ratios of the solubility in water to that in the mixed solvents
listed in Table 2. calculated by Eq. (3) with parameters of Table 2 are shown in Fig. 5.
Root-mean-square deviation, rmsd, is dened as follows: While R = 20 or R = 40, the ratio almost keeps constant along with
 1/2 temperature.
1
N
Enthalpy and entropy of dissolution were studied. The vant Hoff
rmsd = (x1 xcalcd )2 (4) equation relates the logarithm of mole fraction of a solute in an
N
i=1 ideal solution as a linear function of the reciprocal of the absolute
From Tables 1 and 2, we can draw the conclusions that the calcu- temperature as follows [9,10]:
lated solubility sets a good coherence with the experimental values.
dis H  S
ln x1 = + dis (5)
CT C
0.008 water
ethanol+water R=10 where dis H and dis S are the enthalpy and the entropy of disso-
ethanol+water R=20 lution, and C is the gas constant. From the experimentally obtained
0.007
ethanol+water R=40 solubility data, plot of ln x1 versus 1/T gives the values of enthalpy
acetone+water R=10 and entropy of dissolution from the slope and the intercept, respec-
0.006 acetone+water R=20 tively. Fig. 6 shows the vant Hoff plot of the logarithm of mole
acetone+water R=40
0.005
fraction solubility versus reciprocal absolute temperature. Table 3
lists the dissolution enthalpy and entropy. From Table 3, it can be
x1

0.004

Table 2
0.003
Parameters of Eq. (3) for l-glutamine in different solvents (R is molar ratio,
R = Nwater /Nacetone or ethanol ).
0.002
Solvents A B C 105 rmsd
0.001 Water 15.944 3319.124 1.773 9.64
Acetone + water, R = 10 19.662 4005.982 2.196 2.53
275 280 285 290 295 300 305 310 315 Acetone + water, R = 20 109.902 2061.911 17.051 3.67
Acetone + water, R = 40 2.591 2655.354 1.039 4.56
T/K
Ethanol + water, R = 10 12.988 2796.316 2.834 1.83
Ethanol + water, R = 20 88.889 7011.483 12.491 3.30
Fig. 4. Mole fraction solubility of l-glutamine in the mixed solvents (R is molar ratio,
Ethanol + water, R = 40 0.262 2866.735 0.757 2.34
R = Nwater /Nacetone or ethanol ).
 Q. Yu et al. / Thermochimica Acta 558 (2013) 69 9

Table 3
Dissolution enthalpy and entropy of l-glutamine in different solvents (R is molar ratio, R = Nwater /Nacetone or ethanol ).

Solvents Water Acetone + water, R = 10 Acetone + water, R = 20 Acetone + water, R = 40

dis H (kJ mol1 ) 23.39 28.22 24.641 24.67

dis S (J mol1 K1 ) 34.47 42.34 34.19 36.42
r2 (coefcient of correlation) 0.9981 0.9979 0.9985 0.9992

Solvents Ethanol + water, R = 10 Ethanol + water, R = 20 Ethanol + water, R = 40

dis H (kJ mol1 ) 30.15 27.74 25.70

dis S (J mol1 K1 ) 49.46 44.76 39.93
r2 (coefcient of correlation) 0.9993 0.9989 0.9997

3.5 ethanol+water R=10 4. Conclusions

ethanol+water R=20
ethanol+water R=40 The solubilities of l-glutamine in mixed solvents
acetone+water R=10 (water + ethanol, water + acetone) are a function of tempera-
3.0
acetone+water R=20
ture, increasing with an increase in temperature. In the mixed
acetone+water R=40
solvent systems, the increase of R increases the solubility, which
xwater/xmixture

2.5 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
2.0 decrease in the same degree. The ratio of the solubility in water
to that in the mixed solvents of R = 20 or R = 40 almost keeps
constant along with temperature. The dissolution enthalpy and
1.5 entropy of l-glutamine in water predicted from the solubil-
ity data using vant Hoff equation is less than that in mixed
solvents.
1.0
280 285 290 295 300 305 310 315
Acknowledgment
T/K
The authors are grateful to Science Foundation of China (NSFC,
Fig. 5. The ratios of the solubility in water to that in the mixed solvents calculated
No. 21106110).
by Eq. (3) with parameters of Table 2 (R is molar ratio, R = Nwater /Nacetone or ethanol ).

References
water
-4.2 ethanol+water R=10
[1] R.R. Van Der Hulst, B. Van Kreel, M. Meyenfeldt, Glutamine and the preservation
-4.5 ethanol+water R=20 of gut integrity, Lancet 341 (1993) 13631365.
ethanol+water R=40 [2] J. Boza Julio, M. Turini, D. Monnoz, F. Montigon, J. Vuichoud, N. Gueissaz,
-4.8 acetone+water R=10 G. Gremaud, E. Pouteau, C. Piguet-Welsch, A. Finot Paul, O. Ballvre, Effect
acetone+water R=20 of glutamine supplementation of the diet on tissue protein synthesis rate of
-5.1
acetone+water R=40 glucocorticoid-treated rats, Nutrition 17 (2001) 3540.
-5.4 [3] I. Kusumoto, Industrial production of l-glutamine, J. Nutr. 131 (2001)
25522555.
lnx1

-5.7 [4] K. Arii, H. Kobayashi, T. Kai, Y. Kokuba, Degradation kinetics of l-glutamine in

aqueous solution, Eur. J. Pharm. Sci. 9 (1999) 7578.
-6.0 [5] J.W. Mullin, Crystallization, 3rd ed., Butterworth-Heinemann, Oxford,
2000.
-6.3
[6] G. Ren, J. Wang, G. Li, Solubility of paroxetine hydrochloride
-6.6 hemi-hydrate in (water + acetone), J. Chem. Thermodyn. 37 (2005)
860865.
-6.9 [7] Q. Yu, X. Ma, L. Xu, Solubility in binary solvent mixtures: pyrene
dissolved in alcohol + toluene mixtures, Thermochim. Acta 551 (2013)
-7.2 7883.
0.0032 0.0033 0.0034 0.0035 0.0036 [8] L.-X. Wang, X.-M. Yu, Q. Xia, F.-B. Zhang, G.-L. Zhang, Solubil-
ity of sodium 4-nitrotoluene-2-sulfonate in (propanol + water) and
K/T (ethylene glycol + water) systems, J. Chem. Thermodyn. 44 (2012)
128132.
Fig. 6. vant Hoff plot of logarithm mole fraction solubility of l-glutamine (R is molar [9] Q. Yu, X. Ma, L. Xu, Determination of the solubility, dissolution enthalpy
ratio, R = Nwater /Nacetone or ethanol ). and entropy of pyrene in different solvents, Fluid Phase Equilib. 319 (2012)
58.
[10] Q. Yu, X. Ma, W. Gao, Determination of the solubility, dissolution enthalpy and
seen that the dissolution enthalpy of l-Gn in water is less than that entropy of suberic acid in different solvents, Fluid Phase Equilib. 330 (2012)
in mixed solvents. 4447.