Article

pubs.acs.org/jced

Dependence of Refractive Index on Concentration and Temperature

in Electrolyte Solution, Polar Solution, Nonpolar Solution, and
Protein Solution
Chan-Yuan Tan and Yao-Xiong Huang*
Department of Biomedical Engineering, Key Laboratory of Biomaterials, Jinan University, Guang Zhou, China

ABSTRACT: The dependence of refractive index on the concen-

tration and temperature in six kinds of aqueous solutions was
investigated. The six solutions were three electrolyte solutions
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(NaCl, KCl, and CaCl2), a polar solution (glucose solution), a

nonpolar solution (ethyl acetate solution), and a protein solution
Publication Date (Web): September 1, 2015 | doi: 10.1021/acs.jced.5b00018

(bovine serum albumin solution). It was found that dn/dc decreased

with temperature and dn/dT decreased with concentration in polar,
nonpolar, and electrolyte solutions. While in protein solution, both
of the derivatives showed an opposite behavior due to the thermal
aggregation effect of proteins. According to the experimental results,
the empirical expressions of the refractive indices in terms of both
concentration and temperature were derived for the six solutions.
By having the derivatives of refractive index n with respect to
concentration c (dn/dc) and with respect to temperature T (dn/dT) respectively from the expressions, the dependence of
(dn/dc) on temperature and that of (dn/dT) on concentration were obtained.

■ INTRODUCTION
Refractive index is one of the most important physical
Table 1. Sample Descriptiona
purity (mass
properties of solutions. By measuring the refractive index of chemical name source fraction)
a binary solution, one can determine the composition of the sodium chloride (NaCl) Guangzhou Chemical Reagent AR (99.5 %)
solution.1−4 So measurements of refractive index are widely Factory
used in many industrial and research applications to determine potassium chloride Tianjin Kermel Chemical Reagents AR (99.5 %)
(KCl) Development Centre
the concentration of solutions. However, refractive index at calcium chloride Guangzhou Chemical Reagent AR (99.5 %)
the same time also varies with temperature, pressure, and wave- (CaCl2) Factory
length.3,5,6 Though in most of the time, by performing the glucose (C6H12O6, Guangzhou Chemical Reagent AR (99 %)
measurement in atmosphere and using specific wavelength, α-D-glucopyranose) Factory
one can eliminate the influences of pressure and wavelength; BSA Guangzhou Rui Special Biological
Technology Co., Ltd.
the effect of temperature should be always considered.
ethyl acetate (C8H16O4) Tianjin DaMao Chemical Factory AR (99 %)
Numerous theoretical and experimental studies have focused a
on the temperature and concentration dependence of the All of the employed samples were of AR purity and did not need
further purification in the experiment.
refractive indices of aqueous solutions.1−3,6,7 The Lorentz−
Lorenz equation is the most often used formula to estimate the
temperature and concentration dependence of the refractive For example, in a number of microbioreactors and microfluid
index. But it does not provide reliable estimation for the devices, it is required to monitor the concentration variation
derivatives of the refractive index with respect to temperature and in some points/regions of a solution by measuring its refractive
concentration. So the values for the coefficients of temperature index. Since in a streamflow, the mass diffusion is usually
and concentration, or the derivatives of the refractive index n with accompanied by thermal diffusion so temperature also changes
respect to temperature T and concentration c: (∂n/∂T) and with time from place to place. Therefore, it is needed to know
(∂n/∂c), have to come from experiments. After obtaining the by what extent the detected refractive index variation reflects
temperature coefficient (AT) or concentration coefficient (Ac), the real change in concentration under temperature fluctuation.
one can give out the empirical expressions for the temperature For this reason, empirical expressions with both the concentration
or concentration dependence of the refractive index. Most of coefficient and the temperature coefficient are necessary for
the empirical expressions available nowadays only give either the
concentration coefficient or temperature coefficient and express Received: January 6, 2015
as n = n0 + AT, or n = n0 + Ac.8,9 However, in some applications, Accepted: August 25, 2015
both temperature and concentration change at the same time.

© XXXX American Chemical Society A DOI: 10.1021/acs.jced.5b00018

J. Chem. Eng. Data XXXX, XXX, XXX−XXX
 Journal of Chemical & Engineering Data Article

binary solutions. They are also important for light scattering as described previously.1,10 Furthermore, whether (∂n/∂T)
experiments of nonequilibrium fluctuations in a liquid mixture and varies with concentration or (∂n/∂c) varies with temperature
can improve the fundamental understanding of binary mixtures is significant in numerous properties and physicochemical
processes in materials such as the Soret coefficient, the glass
Table 2. Experimental Values of Refractive Index (nD) at transition temperature, and the thermal lens effect of
Temperature T, Mass Fraction w (kg/kg), and Pressure solutions.11−13
p = 0.1 MPa for the Solutions of Glucosea Accordingly in this study, we investigated the simultaneous
nD dependence of refractive index on concentration and temper-
ature in some typical aqueous solutions. The investigated
w 293.15 K 298.15 K 303.15 K 308.15 K 313.15 K 318.15 K
solutions include electrolyte solution, polar solution, nonpolar
0.01 1.3360 1.3355 1.3349 1.3345 1.3337 1.3329 solution, and protein solution. We deduced the empirical
0.05 1.3412 1.3406 1.3401 1.3395 1.3388 1.3379 expressions which include both the concentration and the
0.10 1.3481 1.3475 1.3469 1.3463 1.3455 1.3447
temperature coefficients for the six solutions, and then
0.15 1.3553 1.3548 1.3542 1.3535 1.3527 1.3519
investigated the dependence of (∂n/∂c) on temperature and
0.20 1.3623 1.3617 1.3610 1.3604 1.3596 1.3588
that of (∂n/∂T) on concentration in the solutions.

■
0.30 1.3775 1.3768 1.3763 1.3757 1.3747 1.3739
0.40 1.3947 1.3939 1.3932 1.3925 1.3919 1.3907
0.50 1.4119 1.4106 1.4100 1.4098 1.4086 1.4073 EXPERIMENTAL PROCEDURE
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a
Standard uncertainties u are u(p) = 10 kPa, u(T) = 0.05 K, u(w) = Three electrolyte solutions (NaCl, KCl and CaCl2) of different
0.0005, and u(nD) = 0.0003. mass fractions (kg/kg, 1 %, 5 %, 10 %, 15 %, 20 %, 25 %) were
Publication Date (Web): September 1, 2015 | doi: 10.1021/acs.jced.5b00018

Figure 1. Refractive indices of the six solutions as functions of solution concentration at different temperatures. (a) sodium chloride; (b) potassium
chloride; (c) calcium chloride; (d) glucose; (e) BSA; (f) ethyl acetate. In each of the six plots, ■, 293.15 K; ●, 298.15 K; ▲, 303.15 K; □, 308.15 K;
○, 313.15 K; △, 318.15 K.

B DOI: 10.1021/acs.jced.5b00018
J. Chem. Eng. Data XXXX, XXX, XXX−XXX
 Journal of Chemical & Engineering Data Article

prepared by dissolving the electrolytes (Guangzhou Chemical Table 3. Experimental Values of Refractive Index (nD)
Reagent Factory, Analytical Reagent) in distilled water. at Temperature T, Mass Fraction w (kg/kg), and Pressure
Glucose solution of different mass fractions (kg/kg, 1 %, 5 %, p = 0.1 MPa for the Solutions of NaCl, KCl, CaCl2, and
10 %, 15 %, 20 %, 30 %, 40 %, 50 %) were prepared by Watera
dissolving the glucose (Guangzhou Chemical Reagent Factory,
nD
AR) in distilled water.
Protein solution of different mass fractions (kg/kg, 0.5 %, w 293.15 K 298.15 K 303.15 K 308.15 K 313.15 K 318.15 K
1 %, 1.5 %, 2 %, 2.4 %, 2.9 %, 3.4 %, 3.8 %) were prepared by NaCl
dissolving bovine serum albumin (BSA, Guangzhou Rui Special 0.01 1.3365 1.3359 1.3354 1.3346 1.3339 1.3333
Biological Technology Co., Ltd.) in distilled water. 0.05 1.3434 1.3428 1.3422 1.3414 1.3407 1.3398
Ethyl acetate solution of different mass fractions (kg/kg, 0.10 1.3521 1.3514 1.3508 1.3502 1.3495 1.3485
0.9 %, 2.7 %, 4.5 %, 6.4 %, 8.2 %) were prepared by dissolving 0.15 1.3610 1.3604 1.3596 1.3590 1.3580 1.3572
ethyl acetate (Tianjin DaMao Chemical Factory, AR) in distilled 0.20 1.3702 1.3692 1.3683 1.3678 1.3670 1.3661
water. 0.25 1.3795 1.3788 1.3780 1.3773 1.3767 1.3757
Table 1 gives the description of the samples used in this KCl
investigation. 0.01 1.3349 1.3345 1.3340 1.3335 1.3330 1.3320
The refractive indices of the solutions at different temper- 0.05 1.3405 1.3401 1.3396 1.3390 1.3384 1.3379
atures, (293.15, 298.15, 303.15, 308.15, 313.15, and 318.15) K, 0.10 1.3486 1.3480 1.3476 1.3470 1.3464 1.3460
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were measured using a AHC5-2WAJ Abbe refractometer with 0.15 1.3577 1.3571 1.3566 1.3560 1.3554 1.3550
a temperature controller (Shanghai Optical Instrument Factory, CaCl2
Publication Date (Web): September 1, 2015 | doi: 10.1021/acs.jced.5b00018

China) at the wavelength of 589.3 nm. The refractometer can 0.01 1.3356 1.3353 1.3349 1.3344 1.3339 1.3331
measure refractive index ranging from 1.3000 to 1.7000 with 0.05 1.3442 1.3437 1.3431 1.3426 1.3420 1.3414
an accuracy of ± 0.0001. It was calibrated using distilled 0.10 1.3568 1.3561 1.3555 1.3551 1.3546 1.3539
water and ethanol before each measurement. Considering all 0.15 1.3689 1.3686 1.3680 1.3671 1.3667 1.3662
the six solutions were aqueous solutions, we also measured 0.20 1.3818 1.3814 1.3807 1.3802 1.3795 1.3789
the temperature dependence of RI in distilled water as listed in 0.25 1.3945 1.3943 1.3940 1.3933 1.3926 1.3920
Table 2 for reference. Water

■ RESULTS AND DISCUSSION

A. Effect of Concentration on the Refractive Index of
a
1.3335 1.3330 1.3324 1.3320 1.3316
Standard uncertainties u are u(p) = 10 kPa, u(T) = 0.05 K, u(w) =
0.0005, and u(nD) = 0.0003.
1.3312

Solution. Figure 1 shows the refractive indices of the six solu-

tions versus solution concentration at different temperatures.
We can see that the refractive indices in all the six solutions
both the concentration and temperature coefficients so that the
increase linearly with concentration. Though with increasing
composition changes of the solution would not be over-
temperature the refractive indices decrease, the slopes of the
estimated by the refractive index variation. By using a data
refractive indices changing with concentration are almost the
analysis program called OriginPro (OriginLab, USA) with the
same at all the temperatures for the six kinds of solutions.
method of ordinary least-squares, we obtained the temperature
Especially in the solutions of sodium chloride, calcium chloride,
and concentration coefficients for all the six kinds of aqueous
and glucose, the variation of temperature from (293.15 to
solutions and established empirical expressions for them as
318.15) K only induce slight decrease of the refractive
follows:
indices. The simulation analysis of the experimental data
shows that, except for the ethyl acetate solution which is of NaCl solution:
the order of 10−4, the concentration coefficients of the
refractive indices in all the other five solutions are of the n = 1.3373 + (1.7682 ·10−3)c − (5.8 ·10−6)c 2
order of 10−3. − (1.3531 · 10−4)(T − 273.15) − (5.1 · 10−8)(T − 273.15)2
Table 2−5 give the detailed data of how the refractive indices (1)
in all of the six solutions vary with mass fraction. A comparison
KCl solution:
of our experimental results with the available literature data
shown in Figure 2 indicates that our data are quite consistent n = 1.3352 + (1.6167 · 10−3)c − (4.0 · 10−7)c 2
(deviation is from 0.0003 to 0.0015 for NaCl solution and just
0.0003 for glucose solution) with those reported previously by − (1.1356 · 10−4)(T − 273.15) − (5.7 · 10−9)(T − 273.15)2
other authors.14−19 (2)
B. Effect of Temperature on the Refractive Index of CaCl2 solution:
Solution. Figure 3 illustrates the dependence of refractive
index on temperature in all of the six solutions at different n = 1.3339 + (2.5067 ·10−3)c − (3.9 ·10−8)c 2
concentrations. It indicates that, in all the solutions, the − (1.1122 · 10−4)(T − 273.15) − (4.0 · 10−8)(T − 273.15)2
refractive indices decrease linearly with temperature in the (3)
range of (293.15 to 318.15) K. All of the temperature
coefficients deduced from the experimental data of the six glucose solution:
solutions are of the order of 10−4. n = 1.3356 + (1.5333 ·10−3)c − (9.0 ·10−5)c 2
C. Empirical Expressions. According to the concentration
and temperature dependences of the refractive indices in the six − (1.2647 · 10−4)(T − 273.15) − (4.0 · 10−8)(T − 273.15)2
kinds of solutions, it is desired to have empirical equations with (4)

C DOI: 10.1021/acs.jced.5b00018
J. Chem. Eng. Data XXXX, XXX, XXX−XXX
 Journal of Chemical & Engineering Data Article

Table 4. Experimental Values of Refractive Index (nD) at Temperature T, Mass Fraction w (kg/kg), and Pressure p = 0.1 MPa
for the Solutions of BSA (Bovine Serum Albumin)a
nD
w 293.15 K 298.15 K 303.15 K 308.15 K 313.15 K 318.15 K
0.005 1.3356 1.3350 1.3345 1.3339 1.3333 1.3323
0.010 1.3372 1.3368 1.3362 1.3354 1.3348 1.3336
0.015 1.3376 1.3373 1.3367 1.3360 1.3352 1.3340
0.020 1.3380 1.3379 1.3374 1.3366 1.3358 1.3351
0.024 1.3388 1.3384 1.3380 1.3373 1.3365 1.3357
0.029 1.3397 1.3392 1.3385 1.3380 1.3374 1.3364
0.034 1.3404 1.3400 1.3396 1.3388 1.3382 1.3375
0.038 1.3411 1.3408 1.3401 1.3397 1.3388 1.3382
a
Standard uncertainties u are u(p) = 10 kPa, u(T) = 0.05 K, u(w) = 0.0005, and u(nD) = 0.0003.

Table 5. Experimental Values of Refractive Index (nD) at Temperature T, Mass Fraction w (kg/kg), and Pressure p = 0.1 MPa
for the Solutions of Ethyl Acetatea
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nD
w 293.15 K 298.15 K 303.15 K 308.15 K 313.15 K 318.15 K
Publication Date (Web): September 1, 2015 | doi: 10.1021/acs.jced.5b00018

0.009 1.3341 1.3335 1.3331 1.3327 1.3321 1.3317

0.027 1.3352 1.3348 1.3342 1.3337 1.3332 1.3326
0.045 1.3368 1.3364 1.3358 1.3353 1.3346 1.3342
0.064 1.3384 1.3380 1.3377 1.3368 1.3358 1.3352
0.082 1.3391 1.3388 1.3383 1.3378 1.3373 1.3365
a
Standard uncertainties u are u(p) = 10 kPa, u(T) = 0.05 K, u(w) = 0.0005, and u(nD) = 0.0003.

Figure 2. Comparison between our results and the reported data about the refractive index of NaCl and glucose solutions at 293.15 K and 298.15 K.
In a, ●, our experimental data of NaCl solution at 293.15 K; ⧫, our experimental data of NaCl solutions at 298.15 K; □, data of NaCl at 298.15 K
from ref 14;14 ◇, data of NaCl at 298.15 K from ref 15;15 ○, data of NaCl at 293.15 K from ref 16.16 In b, ⧫, ▲, our experimental data of aqueous
glucose solution at 293.15 K and 298.15 K, respectively; ○, data of glucose at 298.15 K from ref 17;17 △, data of glucose at 298.15 K from ref 18;18
◇, data of glucose at 293.15 K from ref 19.19 The dashed lines in (a) and (b) were plotted according to eqs 1 and 4, respectively for 293.15 K,
whereas the solid line in (a) and (b) were plotted according to eqs 1 and 4, respectively for 298.15 K.

protein solution: of 10−4. In a comparison of the six equations with the Lorentz−
Lorenz equation:20
n = 1.3384 + (1.5985 · 10−3)c + (3.1 · 10−5)c 2
n2 − 1 1 NA
− (1.3939 · 10−4)(T − 273.15) + (2.4 · 10−7)(T − 273.15)2 2
= α(ρ , T , λ),
(5) n +2ρ 3ε0M (7)

ethyl acetate solution:
n = 1.3360 + (0.7125 · 10−3)c − (2.6 · 10−6)c 2
− (1.1566 · 10−4)(T − 273.15) − (1.7 · 10−7)(T − 273.15)2
(6)
where c is the concentration of the solution and T is tempera-
ture in Kelvin. The concentrations in all the equations are in
kg/kg·100. We can see that most of the concentration co-
efficients (except the ethyl acetate solution) are in the order of
10−3, whereas all the temperature coefficients are in the order
D DOI: 10.1021/acs.jced.5b00018
it was proven that the result obtained from each of them was
highly consistent with that of the Lorentz−Lorenz equation
(see Figure 4).
D. Variation of dn/dc with Temperature. Figure 5 shows
the variation of dn/dc vs temperature. We can see that, in all
the six solutions, the derivatives of the refractive index n with
respect to concentration just slightly change with temperature
and the depending coefficients are in the order of 10−6−10−8.
Except that of the protein (BSA) solution, in all the other five
solutions, no matter it is polar, nonpolar, or electrolyte solution,
dn/dc decreases with temperature. The reason for the protein
J. Chem. Eng. Data XXXX, XXX, XXX−XXX
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Publication Date (Web): September 1, 2015 | doi: 10.1021/acs.jced.5b00018

Figure 3. Dependence of refractive index on temperature in all the six solutions at different concentrations. (a) Sodium chloride; (b) potassium
chloride; (c) calcium chloride; (d) glucose; (e) BSA; (f) ethyl acetate.

Figure 4. Fitting curves of our empirical equation (solid line) and the Lorentz−Lorenz equation (dashed line) for NaCl solution. Dots are the
experimental data. The left one is the RI variation with concentration at 293.15 K; the right one is the RI variation with temperature at 5%
concentration. Both of the curves fit with the experimental point quite well. Similar results were also found in the other five solutions.

solution to have an opposite behavior is probably due to the the protein aggregates to increase with temperature,21,22 so the
structural variation of the protein with temperature. For there is dn/dc of the solutions increases with temperature. Since protein
a thermal aggregation effect on BSA to lead the particle radii of thermal aggregation is quite common in protein solutions, it is
E DOI: 10.1021/acs.jced.5b00018
J. Chem. Eng. Data XXXX, XXX, XXX−XXX
 Journal of Chemical & Engineering Data
Figure 5. Variation of dn/dc vs temperature.
expected that the dn/dc of other protein solutions would also
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increase with temperature.
As mentioned above that, the refractive index increment,
Publication Date (Web): September 1, 2015 | doi: 10.1021/acs.jced.5b00018
dn/dc is significant in polymer solutions for determining mole-
cular weight with the Zimm equation23 using the technique of
static light scattering. Therefore, it is needed to know if the
variation of dn/dc with temperature in the protein solution is
within an acceptable range. According to our experimental
results for the solutions, there were only very little changes in
the dn/dc with temperature in all the six solutions so that they
can be neglected.
E. Variation of dn/dT with concentration. The gradient
of refractive index with respect to temperature dn/dT, is also a
Figure 6. Variation of dn/dT vs concentration. (a) ⧫, NaCl solution;
solution; (d) ethyl acetate solution.
Article
significant parameter in various applications, such as in the
measurements of the glass transition temperature13 and thermal
lens effect.11 Whether dn/dT depends on concentration should
be concerned for these applications. Figure 6 illustrates the varia-
tion of dn/dT vs concentration. Similar to the case of dn/dc
varying with temperature, except the protein solution, in all the
other five solutions, dn/dT just slightly decreases (or increases
in absolute value) with concentration, and the depending
coefficients are in the order of 10−7−10−8 thus can be neglected
in most cases. However, in the protein solution, dn/dT increases
with concentration and the depending coefficient is in the order
of 10−6. This phenomenon is also probably due to the thermal
aggregation effect of proteins.
■ CONCLUSIONS
In this paper, we investigated the temperature and concentration
dependences of refractive indices of six aqueous solutions and
established empirical expressions with both concentration
coefficient and temperature coefficient for the solutions. We also
deduced the variation of dn/dc vs temperature and the variation of
dn/dT vs concentration for the six solutions. We demonstrated
that dn/dc decreases with temperature and dn/dT decreases with
concentration in polar, nonpolar, and electrolyte solutions. While in
protein solution, both of the derivatives show an opposite behavior
due to the thermal aggregation effect of proteins. These findings
can help to improve the fundamental understanding of binary
mixtures, and are significant for the determination of concentration,
the Soret coefficient, the glass transition temperature, the molecular
weight, and the thermal lens effect of solutions.
■, KCl solution and ▲, CaCl2 solution; (b) glucose solution; (c) protein
F DOI: 10.1021/acs.jced.5b00018
J. Chem. Eng. Data XXXX, XXX, XXX−XXX
 Journal of Chemical & Engineering Data

■
Article

AUTHOR INFORMATION (16) O'Donnell, P. B.; McGinity, J. W. Preparation of microspheres

Corresponding Author
*Phone number: +86-20-85223742. Fax number: +86-20-
85223742. E-mail address: tyxhuang@jnu.edu.cn.
Present Address
(C-Y.T. and Y.-X.H.) Department of Biomedical Engineering,
Ji Nan University, Guang Zhou, China 510632.
Funding
This work was supported partly by the Chinese National
Natural Science Foundation (Nos. 30940019 and 60377043)
and Guang Dong Provincial Science and Technology
Foundation.
Notes
The authors declare no competing financial interest.
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G DOI: 10.1021/acs.jced.5b00018
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