Thermochimica Acta 575 (2014) 1216

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

Thermodynamic properties of vitamin B2

A.V. Knyazev , I.A. Letyanina, A.S. Plesovskikh, N.N. Smirnova, S.S. Knyazeva
Lobachevsky State university of Nizhni Novgorod, Gagarin Prospekt 23/2, 603950 Nizhni Novgorod, Russia

a r t i c l e

i n f o

Article history:
Received 7 August 2013
Received in revised form
21 September 2013
Accepted 25 September 2013
Available online 24 October 2013
Keywords:
Vitamin B2
Riboavin
Adiabatic vacuum calorimetry
Heat capacity
Combustion calorimetry
Thermodynamic functions

a b s t r a c t
In the present work temperature dependence of heat capacity of vitamin B2 (riboavin) has been measured for the rst time in the range from 6 to 322 K by precision adiabatic vacuum calorimetry. Based
on the experimental data, the thermodynamic functions of the vitamin B2 , namely, the heat capacity,
enthalpy H (T) H (0), entropy S (T) S (0) and Gibbs function G (T) H (0) have been determined for
the range from T 0 to 322 K. The value of the fractal dimension D in the function of multifractal generalization of Debyes theory of the heat capacity of solids was estimated and the character of heterodynamics
of structure was detected. In a calorimeter with a static bomb and an isothermal shield, the energy of
combustion of the riboavin has been measured at 298.15 K. The enthalpy of combustion c H and the
thermodynamic parameters f H , f S , f G and of reaction of formation of the riboavin from simple
substances at T = 298.15 K and p = 0.1 MPa have been calculated.
Published by Elsevier B.V.

1. Introduction
Riboavin (PubChem CID: 493570), also known as vitamin B2
is an easily absorbed colored micronutrient with a key role in
maintaining health in humans and animals. It is the central component of the cofactors avin adenine dinucleotide (FAD) and avin
mononucleotide (FMN), and is therefore required by all avoproteins. The current recommended dietary allowances (RDA) for
riboavin for adult men and women are 1.3 mg/day and 1.1 mg/day,
respectively [1]. Riboavin has been used in several clinical and
therapeutic situations. For over 30 years, riboavin supplements
have been used as part of the phototherapy treatment of neonatal
jaundice. The light used to irradiate the infants breaks down not
only bilirubin, the toxin causing the jaundice, but also the naturally
occurring riboavin within the infants blood, so extra supplementation is necessary [2]. Various biotechnological processes
have been developed for industrial scale riboavin biosynthesis
using different microorganisms, including lamentous fungi such
as Ashbya gossypii, Candida famata and Candida averi, as well as
the bacteria Corynebacterium ammoniagenes and Bacillus subtilis
[3].
The goals of this work include calorimetric determination of
the standard thermodynamic functions of the riboavin with
the purpose of describing biochemical and industrial processes

Corresponding author. Tel.: +7 831 462 32 34; fax: +7 831 434 50 56.
E-mail address: knyazevav@gmail.com (A.V. Knyazev).
0040-6031/$ see front matter. Published by Elsevier B.V.
http://dx.doi.org/10.1016/j.tca.2013.09.032

with it participation. These data can be used to evaluate the

thermodynamic properties of avin mononucleotide (FMN) and
avin adenine dinucleotide (FAD).
These data can be used to evaluate the thermodynamic
properties of avin mononucleotide (FMN) and avin adenine dinucleotide (FAD).
2. Experimental
2.1. Sample
Riboavin was purchased from Fluka. For phase identication,
an X-ray diffraction pattern of the vitamin B2 sample was recorded
on a Shimadzu X-ray diffractometer XRD-6000 (CuK radiation,
geometry 2) in the 2 range from 5 to 60 with scan increment
of 0.02 (Fig. 1). The X-ray data and estimated impurity content
(0.1 wt%) in the substance led us to conclude that the riboavin
sample studied was an individual crystalline compound (modication I [4]).
2.2. Apparatus and measurement procedure
To measure the heat capacity Cpo of the tested substance in
the range from 6 to 330 K a BKT-3.0 automatic precision adiabatic vacuum calorimeter with discrete heating was used. The
calorimeter design and the operation procedure were described
earlier [5]. The calorimeter was tested by measuring the heat
capacity of high-purity copper and reference samples of synthetic

A.V. Knyazev et al. / Thermochimica Acta 575 (2014) 1216

Table 1
Experimental values of isobaric heat capacities (J K1 mol1 ) of crystalline riboavin;
M = 376.3682 g mol1 .

I, pulses
8000

6000

4000

2000

2,

0
5

13

10

15

20

25

30

35

40

45

50

55

60

Fig. 1. X-ray diffraction patterns for riboavin (modication I).

corundum and K-2 benzoic acid. The analysis of the results showed
that measurement error of the heat capacity of the substance at
helium temperatures was within 2%, then it decreased to 0.5%
as the temperature was rising to 40 K, and was equal to 0.2% at
T > 40 K.
The energy of combustion, c U, of riboavin was measured in
a calorimeter (V-08) with a static bomb and an isothermal shield.
The calorimeter design, the procedure of measuring the energies of
combustion and the results of calibration and testing are given elsewhere [6]. It should be noted that while checking the calorimeter
by burning succinic acid, prepared at D.I. Mendeleev Research Institute of Metrology (the value of the standard enthalpy of combustion
of the acid coincided with the certicate value within 0.017%). For
complete combustion of riboavin we used parafn as an auxiliary
substance.
3. Results and discussion

T (K)

Cpo

T (K)

Cpo

T (K)

Cpo

Series 1
6.75
7.32
8.41
8.94
9.34
9.68
10.15
10.36
10.98
11.56
11.98
13.16
15.03
16.92
18.77
20.65
22.79
24.95
27.13
29.37
31.61
33.91
36.25
38.52
40.96
43.38
45.72
48.12
207.64
210.68
213.72
216.77
219.81
222.86
225.93
228.98
232.02
235.06
238.11
241.15
244.20

2.55
2.83
3.45
4.08
4.48
4.81
5.25
5.42
6.23
7.05
7.68
9.25
12.22
15.24
18.25
21.38
25.35
29.29
33.81
38.36
42.49
47.31
51.78
56.45
60.98
65.91
70.28
74.84
317.5
321.4
325.8
330.0
334.0
338.3
342.3
346.6
350.4
355.0
359.4
363.5
367.7

50.55
52.99
55.45
57.93
60.48
62.91
65.40
67.89
70.37
72.85
75.32
77.79
80.34
82.75
85.22
88.86
Series 2
83.48
86.72
89.48
92.71
95.70
98.70
101.67
104.65
107.64
110.63
113.62
116.63
247.25
250.29
253.34
256.38
259.42
262.46
265.50
268.54
271.58
274.65
277.69
280.72
283.75

79.52
83.98
88.71
93.42
98.18
102.6
106.9
110.9
115.1
119.2
123.3
128.2
132.4
137.1
141.4
147.9

119.63
122.64
125.66
128.69
131.70
134.72
137.75
140.77
143.80
146.83
149.86
152.90
155.94
158.98
162.02
165.06
168.10
171.14
174.19
177.26
180.30
183.34
186.38
189.42
192.46
195.50
198.54
201.58
204.61
286.78
289.80
292.82
295.84
298.86
302.37
306.36
310.36
314.34
318.32
322.29

194.3
198.5
202.7
207.3
211.6
215.9
220.3
224.6
229.0
233.0
237.2
241.7
246.0
250.1
254.2
258.6
263.0
267.3
271.6
276.0
279.9
283.9
288.1
291.9
296.2
300.5
305.0
308.7
313.0
425.5
429.8
434.0
438.2
442.2
447.3
453.1
459.1
464.5
470.0
474.7

138.3
144.2
148.9
153.8
158.4
162.9
167.8
172.4
177.1
181.6
185.8
190.1
371.5
375.6
379.8
383.9
388.4
392.5
396.6
400.6
404.8
409.2
413.5
417.4
421.5

3.1. Heat capacity

The Cpo measurements were carried out between 6 and 322 K
(Table 1). The mass of the sample loaded in the calorimetric
ampoules of the BKT-3.0 device was 0.3779 g. 122 experimental Cpo
values were obtained in two series of experiments. The heat capacity of the sample varied from 20% to 50% of the total heat capacity
of calorimetric ampoule + substance over the range between 6 and
322 K. The experimental points of Cpo in the temperature interval
20322 K were tted by means of the least-squares method and
polynomial equations (Eqs. (1)(2)) of the Cpo versus temperature
have been obtained. The corresponding coefcients (A, B, C, etc.)
are given in Table 2.
Cpo

= A+B
+F

T 
30

 T 5
30

+C

+G

 T 2
30

 T 6
30

+D

+H

 T 3
30

+E

 T 4
30
(1)

30

+ F ln5 (T/30) + G ln6 (T/30) + +H ln7 (T/30) + I ln8 (T/30)

10

11

Cp, JK-1mol-1
500

400

 T 7

Cpo = A + B ln(T/30) + C ln2 (T/30)+D ln3 (T/30) + E ln4 (T/30)

Their root mean square deviation from the averaging Cpo = f (T )

curve was 0.15% in the range T = (640) K, 0.075% from T = (40
to 80) K and 0.050% between T = (80 and 322) K. The experimental
values of the molar heat capacity of riboavin over the range from 6
to 322 K and the averaging Cpo = f (T ) plot are presented in Fig. 2. The
heat capacity Cpo of this substance gradually increases with rising
temperature and does not show any peculiarities.

300

200

100

12

+ J ln (T/30) + K ln (T/30) + L ln (T/30) + M ln (T/30)

+ Q ln16 (T/30)

T, K

+ + N ln13 (T/30) + O ln14 (T/30) + P ln15 (T/30)

(2)

50

100

150

200

250

300

Fig. 2. Temperature dependence of heat capacity of riboavin.

350

14

A.V. Knyazev et al. / Thermochimica Acta 575 (2014) 1216

Table 2
Coefcients in the tting polynomials for riboavin.
T (K)
Polynomial type

2888
2

84228
2

200322
1

A/J
B/K
C/L
D/M
E/N
F/O
G/P
H/Q
I

39.496328/237990.48
57.541844/262521.52
30.626419/1213173.1
176.57956/1435044.3
496.96487/559655.31
3283.224/239150.84
15934.76/278304
7230.3208/68815.08
152620.33

24110.87/5401.426
97414.132/560.58502
134767.007
34595.47
98263.138
101594.62
18493.249
26533.786
19533.76

650472.6351
541748.6477
192521.8858
37832.58703
4440.40908
311.2899925
12.06941506
0.1996650593

From the experimental Cpo values in the range 2040 K the value
of the fractal dimension D of the riboavin was evaluated. According to the fractal theory of the heat capacity [7], D is the most
important parameter that species the character of heterodynamics of the substance structure. For solids of a chain structure the
relation Cpo versus T at lower temperatures is proportional to T1 ,
of a layer structure to T2 and of steric one to T3 [8]. In the fractal
theory of the heat capacity, an exponent on T is the heat capacity

function is denoted by D and is called the fractal dimension. This

follows specically from Eq. (3) [7]:

 T D

Cv = 3D(D + 1)kN(D + 1)(D + 1)

max

(3)

where N is the number of atoms in a formula unit, k the Boltzmann

constant, (D + 1) the -function, (D + 1) the Riemann -function,
and  max is the characteristic temperature. As follows from

Table 3
Thermodynamic functions of crystalline riboavin; M = 376.3682 g mol1 , p = 0.1 MPa.
T (K)

Cpo (T ) (J K1 mol1 )

H (T ) H (0) (kJ mol1 )

S (T ) (J K1 mol1 )

[G (T ) H (0)] (kJ mol1 )

0
1
2
3
4
5
6
7
8
9
10
15
20
25
30
35
40
45
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
273.15
280
290
298.15
310
320
322

0
0.0523
0.209
0.470
0.836
1.310
1.880
2.650
3.250
4.080
5.100
12.00
20.35
29.50
39.49
49.35
59.26
68.91
78.34
97.31
114.4
132.0
149.6
165.2
180.4
195.0
209.2
223.3
237.5
251.7
265.7
279.4
293.0
306.7
320.5
334.7
348.2
361.5
375.1
389.1
403.0
407.4
416.6
429.9
441.1
458.6
472.1
474.1

0
0.000020
0.000100
0.000500
0.00110
0.00220
0.00380
0.00600
0.00900
0.0125
0.0172
0.05920
0.1398
0.2636
0.4363
0.6584
0.9300
1.251
1.619
2.497
3.558
4.788
6.196
7.771
9.499
11.38
13.40
15.56
17.86
20.31
22.90
25.62
28.48
31.48
34.62
37.89
41.31
44.86
48.54
52.36
56.32
57.60
60.42
64.65
68.20
73.53
78.19
79.13

0
0.0261
0.105
0.235
0.418
0.653
0.941
1.280
1.660
2.100
2.580
5.898
10.48
15.97
22.23
29.06
36.30
43.84
51.59
67.55
83.87
100.3
116.8
133.4
149.9
166.2
182.4
198.4
214.3
230.1
245.7
261.3
276.8
292.2
307.5
322.7
337.9
353.0
368.0
383.0
397.9
402.6
412.8
427.7
439.8
457.3
472.1
475.0

0
0.00000900
0.0000700
0.000235
0.000558
0.00109
0.00188
0.00299
0.00433
0.00634
0.00863
0.02926
0.06971
0.1355
0.2307
0.3587
0.5220
0.7222
0.9607
1.556
2.313
3.234
4.319
5.570
6.987
8.567
10.31
12.21
14.28
16.50
18.88
21.41
24.10
26.95
29.95
33.10
36.40
39.86
43.46
47.22
51.12
52.38
55.17
59.38
62.91
68.23
72.87
73.82

A.V. Knyazev et al. / Thermochimica Acta 575 (2014) 1216

15

Table 4
Experimental data on combustion energy for riboavin at T = 298.15 K.
Value

Experiment
1

msam (g)a
mpar (g)a
mthread (g)a
W (J g1 )b
R + (R)c ()
c U d (J)

0.07221
0.3149
0.0065
76,057
0.215552
16394.2

0.06398
0.2615
0.0059
76,057
0.180295
13712.6

0.06238
0.3131
0.0052
76,057
0.211404
16078.7

0.04761
0.1714
0.0051
76,057
0.119977
9124.9

0.06652
0.2387
0.0051
76,057
0.166798
12686.3

0.06814
0.2401
0.0050
76,057
0.168091
12784.2

c Upar e (J)
c Uthread e (J)
c UHNO3 f (J)
g (J)
c Uh (J g1 )

14719.7
108.8
6.4
17.3
21594.0

12228.2
98.7
3.3
17.3
21606.6

14635.2
87.0
7.5
17.3
21625.0

8011.9
85.4
4.1
17.2
21498.2

11157.9
85.4
7.5
17.3
21581.1

11223.2
83.7
6.7
17.3
21582.3

c U = 21, 581 36 J g1 = 8121 13 kJ mol , the mean energy of combustion of riboavin.

1
c U = 8116 13 kJ mol , the energy of combustion of riboavin at standard pressure.
a
msam , mpar , mthread , masses of the tested sample, parafn and a cotton thread, respectively.
b
W, the energy equivalent of the calorimeter.
c
R + (R), the change in resistance of platinum-resistance thermometer with correction for heat-exchange.
d
c U , the total energy released during the experiment.
e
f
g
h

c Upar , c Uthread , amounts of energy released on burning parafn and cotton thread, respectively.
c UHNO3 , the energy of formation of nitric acid.
, Washburn correction.
c U, the energy released on burning of riboavin.

Table 5
Enthalpy of combustion and thermodynamic characteristics of formation of riboavin (T = 298.15 K, p = 0.1 MPa).
Compound

c H (kJ mol1 )

f H (kJ mol1 )

f S (J K1 mol1 )

f G (kJ mol1 )

C17 H20 N4 O6

8116 13

1432 13

1961 3

847 14

inferences [7], D can be evaluated from the experimental data on

the temperature-dependent heat capacities from a slope of the corresponding rectilinear sections of the plot ln Cv versus ln T. Without
a substantial uncertainty it may be assumed that at T < 50 K Cpo = Cv .
From the ln Cv versus ln T plot and Eq. (3) it was found that in the
range 2040 K, D = 2,  max = 182.6 K for riboavin. With these values
of D and  max Eq. (3) reproduces the experimental Cpo values in the
temperature range mentioned with an uncertainty of 0.93%. The
D-value points to the layer structure of riboavin [810].

3.2. Thermodynamic functions

To calculate the standard thermodynamic functions (Table 3) of
riboavin, Cpo values of it were extrapolated from the starting temperature of the measurement (approximately 7 K) to 0 K by Debyes
function of heat capacity [11]:


Cpo = nD

D
T

3.3. Thermodynamic characteristics of riboavin formation

The experimental data on burning of riboavin are presented in
Table 4. As a result, the energies and enthalpies of combustion of
riboavin at T = 298.15 K and standard pressure were determined.
The values are for the reaction:
C17 H20 N4 O6 (cr) + 19O2 (g) 17CO2 (g) + 10H2 O(l) + 2N2 (g)
In brackets are given the physical states of reagents: (cr), crystalline; (g), gaseous; (l), liquid.
The data on the enthalpy of combustion of the crystalline
riboavin was used to estimate enthalpy of combustion and formation at T = 298.15 K and p = 0.1 MPa. From the absolute value

Q, Wg-1

12

exo

(4)

10

where D is the symbol of Debyes function, n = 3 and

 D (C17 H20 N4 O6 ) = 83.0 K are specially selected parameters [11].
Eq. (4) with the above parameters describes the experimental Cpo
values of the compound between 7 and 10 K with the error of
2.15%.
In calculating the functions it was assumed that Eq. (4) reproduces the Cpo values of riboavin at T < 7 K with the same error. The
calculations of H o (T ) H o (0) and S o (T ) S o (0) were made by the
numerical integration of Cpo = f (T ) and Cpo = f (ln T ) curves, respectively, and the Gibbs function Go (T ) H o (0) was estimated from the
enthalpies and entropies at the corresponding temperatures. The
error of the function values was 1% at T < 40 K, 0.5% between 40
and 80 K, 0.2% in the range from 80 to 322 K.

2
Tm(dec)=571 K
T, K

0
540

550

560

570

Fig. 3. DSC curve of riboavin.

580

590

16

A.V. Knyazev et al. / Thermochimica Acta 575 (2014) 1216

of the entropy of the riboavin (Table 3), carbon in the form

of graphite, gaseous hydrogen, oxygen and nitrogen [12,13] the
standard entropy of formation f S of riboavin at T = 298.15 K was
calculated by methods described earlier [14]. The Gibbs function
of formation f G of the riboavin was evaluated from the f H
and f S values (Table 5). The values conform to the following
process:
17C(gr) + 10H2 (g) + 2N2 (g) + 3O2 (g) C17 H20 N4 O6 (cr)
where in the brackets are indicated the physical states of reagents:
(gr), graphite; (g), gaseous; (cr), crystalline.
3.4. Differential scanning calorimetry
Thermal analysis made it possible to establish some peculiarities of processes taking place in the compound under investigation
during heating. Fig. 3 represents DSC curve of riboavin, where we
can see endothermic effect at 571 K. This effect is connected with
melting and decomposition of compound.
4. Conclusions
The general aim of these investigations was to report the results
of the thermodynamic study of the riboavin. The heat capacity of this vitamin B2 is measured in the temperature range from
6 to 322 K, the thermodynamic functions are calculated and the
fractal dimension D is evaluated. Thermochemical parameters of
formation are determined by combining the data obtained by using
combustion calorimetry and heat capacity measurements.

Acknowledgement
The work was performed with the nancial support of the Russian Foundation of Basic Research (Project Number 13-03-00152).
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