Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 1397–1402

Contents lists available at ScienceDirect

Spectrochimica Acta Part A: Molecular and

Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa

FT-IR spectroscopic, thermal analysis of human urinary stones

and their characterization
R. Selvaraju ⇑, A. Raja, G. Thiruppathi
Department of Engineering Physics, Annamalai University, Annamalai Nagar 608 002, Tamil Nadu, India

h i g h l i g h t s g r a p h i c a l a b s t r a c t

 The chemical compounds were

identified by FT-IR technique.
 Urinary stone minerals were
identified by XRD study.
 Endothermic and exothermic peaks
were identified by TG–DTA analyses.

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

Article history: In the present study, FT-IR, XRD, TGA–DTA spectral methods have been used to investigate the chemical
Received 7 June 2014 compositions of urinary calculi. Multi-components of urinary calculi such as calcium oxalate, hydroxyl
Received in revised form 25 August 2014 apatite, struvite and uric acid have been studied. The chemical compounds are identified by FT-IR spec-
Accepted 18 September 2014
troscopic technique. The mineral identification was confirmed by powder X-ray diffraction patterns as
Available online 28 September 2014
compared with JCPDS reported values. Thermal analysis techniques are considered the best techniques
for the characterization and detection of endothermic and exothermic behaviors of the urinary stones.
Keywords:
The percentages of each hydrate (COM and COD) are present together, in the presences of MAPH or
Urinary calculi
FT-IR
UA. Finally, the present study suggests that the Urolithiasis is significant health problem in children,
XRD and is very common in some parts of the world, especially in India. So that present study is so useful
TGA–DTA and helpful to the scientific community for identification of latest human health problems and their rem-
edies using spectroscopic techniques.
Ó 2014 Elsevier B.V. All rights reserved.

Introduction
Urolithiasis is a very common disease in both developed and
underdeveloped countries and its incidence is increasing world-
wide. It is fraught with high recurrence and is associated with sig-
nificant morbidity. Some studies report that approximately 20% of
the world’s population suffers from urinary stone disease [1]. In
⇑ Corresponding author. Mobile: +91 9994784685.
E-mail address: drselvarajufeatau@gmail.com (R. Selvaraju).
India, urolithiasis affects about 2 million people every year [2].
The oldest renal stone on record was described by Shattock in
1905 and was found in an Egyptian mummy in a tomb dating to
approximately 4400 BC [3]. Stone analysis is of great importance
to the recurrence rate for medically treated urolithiasis is >50% at
5 years [4]. Although urolithiasis is usually considered a disease
of adults, it also occurs in children [5–7]. In addition, recurrence
rates are higher in children than in adults. The possible reason
may be related to metabolic disorders [8]. A renal calculus is a solid
concretion (or) crystal aggregation formed in the kidney from

http://dx.doi.org/10.1016/j.saa.2014.09.046
1386-1425/Ó 2014 Elsevier B.V. All rights reserved.
1398 R. Selvaraju et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 1397–1402
dietary minerals in the urine. Urinary stones are typically classified
by their location in the kidney, urethra, and bladder. Kidney stones
are hard, rock-like crystals of varying sizes and shapes. These occur
when salts in the urine precipitate and form solid materials. Uri-
nary stones may contain various combinations of chemicals [9].
The epidemiology of pediatric urinary stone disease has hitherto
not been as rigorously defined as that in adults. The factors affect-
ing adult litho genesis may include diverse climate, racial mix,
nutritional habits, lifestyle, obesity [10] and urinary abnormality,
metabolic disorders, and chronic urinary tract infection. Urolithia-
sis is a significant health problem in children, and is very common
in some parts of the world, especially in India. Most of these pedi-
atric patients are also considered to be at high risk for recurrence
[11]. Stone analysis is great importance to the therapy and meta-
phylaxis of residual and recurrent stones [12,13]. The recurrent
pathology may be related to the functional and morphologic
changes in the urinary tract, environmental factors, urogenital
abnormalities, and metabolic disorders [14]. Therefore, prevention
of stone formation is of utmost importance. Under these circum-
stances, prevention, in terms of practicality, means medical treat-
ment [15]. Accurate analysis of urinary stone composition is a
key factor for choosing the most appropriate medical management.
Climatic, dietary, genetic, and socioeconomic factors have been
related to geographic variations observed in the incidence, compo-
sition, and clinical characteristics of urinary stone disease [16]. In
FT-Raman, the shift in wavenumber depends upon the chemical
structure of the molecules those are responsible for scattering.
Raman spectroscopic studies of the composition of urinary calculi
use the spectra of the different chemical components were
reported in literatures [17–21]. According to previous studied, it
is cleared that the FT-IR and FT-Raman spectral of human urinary
stones were identified the chemical compositions such as calcium
oxalate, calcium phosphate, struvite and uric acid [22,23]. In the
present study, qualitative analyses of human urinary stones and
the characteristic peaks for dehydration and decomposition of cal-
cium oxalate, struvite, uric acid and calcium bilirubinate, calcium
phosphate or hydroxyapatite up to 800 °C have been reported.
Experimental details
The 10 urinary stone samples (KS1–KS10) were collected from
the Department of Urology, Rajah Muthiah Medical College Hospi-
tal (RMMC&H), Annamalai University, Annamalai Nagar, Tamil
Nadu, South India. The FT-IR spectra of urinary stones are recorded
using KBr pellet technique in the region of 4000–400 cm1. The FT-
IR measurements were performed using a Perkin Elmer Spectrum
RX1 Model FT-IR spectrometer available at Centralized Instrumen-
tation of Service Laboratory, Department of Physics, Annamalai
University, Tamil Nadu, South India.
The XRD patterns of the well powdered samples were charac-
terized by powder X-ray diffraction analysis using X-ray diffrac-
tometer model Rigaku D/max-2500 with monochromatic Cu Ka
source (k = 1.54056 Å) using tube voltage and current of 40 kV
and 100 mA, respectively. The samples are scanned over the range
(2h) 10–80°. The X-ray diffraction of patterns of the urinary stone
samples are obtained at Rigaku D/max-2500 XRD diffractometer
available at University of Madras, Guindy campus, Chennai, Tamil
Nadu, South India.
Thermogravimetric analyses were obtained under nitrogen
atmosphere at a heating rate 8 °C min1 in Pt cell and standard
gas flow on TA Instruments Model SDT Q600 apparatus. The tem-
perature range from 25 °C to 800 °C in up scan direction. In the
present study, thermal studies are carried out in TA instrument
model SDT D600, available at Nano science and technology, Anna
University, Guindy campus, Chennai, Tamil Nadu, South India.
Results and discussion
FT-IR spectra of ten urinary stones samples are qualitatively
analyzed. Samples were numbered sequentially KS1 to KS10. The
major constituents of calcium oxalate monohydrate and calcium
phosphate was found. The presence of calcium oxalate monohy-
drate in the urinary stone is characterized by a large symmetric
and asymmetric OAH stretching absorption band between 3493
and 3059 cm1 [24]. The strong absorption band observed at
2931 cm1 belongs to vibration of CAH stretching [25]. The spec-
trum of oxammite (ammonium oxalate monohydrate) shows
absorption bands observed at 1905 cm1 stretching vibration and
C@O stretching vibration of oxammite [26]. The strong absorption
bands are observed at 1658 cm1 corresponding to the NAH defor-
mation [24]. The strong absorption bands at 1612, 1625 cm1 cor-
responding to the vibration of C@O stretching of COM. The weak
absorption bands at 1375 cm1 belongs to vibration of CAO
stretching. The strong absorption bands at 1319 cm1 correspond-
ing to the vibration of CAO stretching [27]. The carbonate of hydro-
xyl apatite vibrational mode showed a peak at 1068 cm1 [28]. The
entire samples exhibit a strong absorption band observed around
1040 cm1, attributed to PAO stretching of calcium phosphate
[29]. The weak absorption band at 945 cm1 is the presence of
CAO stretching [24]. Strong absorption band observed at
884 cm1 belongs to CAC stretching. The bands at 779 and
665 cm1 are corresponding to the CAC stretching and the out of
plane OAH bending [23]. The discrete formation of the two bands
at 779 cm1 and 516 cm1 are important for distinguishing COM
from COD [30]. In addition to the above, calcium phosphate has
characteristic bands at 604 and 569 cm1, which belongs to PO3 4
bending vibration. The absorption wave numbers of COM and apa-
tite with their corresponding tentative assignments are presented
in Table 1 (see Fig. 1).
In group II, five urinary stone samples were found to be a mix-
ture of calcium oxalate monohydrate (COM), calcium phosphate
(apatite), magnesium ammonium phosphate (MAPH) and uric acid
with the major constituents of apatite and MAPH and the minor
constituents of COM and uric acid. The broad band at 3403–
2918 cm1 the presence of OAH group [26]. Strong absorption
bands is observed at 1655 cm1 corresponding to the Vibration
of C@O respectively [24]. Strong and broad band extending from
979 and 904 cm1 observed in group-II sample with corresponding
to PO43 stretching vibration [31]. It indicates the absorption of
PO3
4 group and by the presence of other bands at 1436,
756 cm1. The peak position and additional bands are observed
at and 464 cm1 are due to NH+4 deformation of uric acid [32].
The absorption frequency of calcium oxalate monohydrate, cal-
cium phosphate, struvite, uric acid, oxammite with their corre-
sponding tentative assignments are presented in Table 2. The
overlap FT-IR spectra of KS5–KS10 are shown in Fig. 2.
XRD analysis
In group urinary stone samples were found to be a mixture of
calcium oxalate monohydrate (COM) and calcium phosphate (Apa-
tite) with major constituents of calcium oxalate monohydrate and
calcium phosphate in minor constituents. The XRD patterns of the
samples under investigation are analyzed; the ‘d’ values of samples
are compared with standard ‘d’ values from JCPDS data. The XRD
pattern (Fig. 3) of calcium oxalate monohydrate (COM) and cal-
cium phosphate (Apatite) with either ‘d’ values and intensities of
experimental values and standard values are shown in Table 3
and the comparing the ‘d’ and I/Io values of COM component pres-
ent in the mixed stone of COM and calcium phosphate (Apatite) are
presented in Table 3. The XRD spectra of KS5 are shown in Fig. 3.
 R. Selvaraju et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 1397–1402 1399

Table 1
FT-IR spectral data of urinary stones (KS1–KS5).

Wavenumber (cm1) Tentative assignments Compounds

KS1 KS2 KS3 KS4 KS5
3493 3493 3493 3487 3487 OAH stretching COM
3342 3342 3432 3336 3340 OAH stretching COM
3243 3251 3265 – 3259 OAH stretching COM
3062 3062 3062 3059 3061 OAH stretching COM
– – – – 2931 CAH stretching COM
– – – 1905 1905 Stretching vibration of C@O Oxammite
– – – 1645 1666 OAH deformation Apatite
– 1625 1622 1637 Vibration of C@O COM
1618 – 1618 – 1612 Vibration of C@O COM
– – – 1375 1379 CAC of vibration COM
1316 1316 1308 1317 1319 Vibration of CAO COM
– – – 1068 – PO3
4 stretching Apatite
1042 1014 1036 1039 1047 PO3
4 stretching Apatite
945 – 945 950 – CAO stretching COM
884 884 876 883 881 CAC stretching COM
771 779 779 779 779 CAC stretch COM
665 665 665 663 665 Out-of-plane OAH bending COM
599 574 604 603 603 PO3
4 stretching Apatite
– – – – 569 PO3
4 bending vibration Apatite
515 515 521 516 516 OACAO in plane bending COM

endothermic process due to water molecule in whewellite at the

first step is estimated to be 13.4%, The weight loss of second step
exothermic peak at the decomposition of carbon dioxide is
observed to be about 16.6%, Weight losses of third endothermic
peaks were 24%. Terminal weight loss is observed to be 58.8% the
external structure of the urinary calculi sample is crystalline which
is characteristic of COD formation. This means that the sample con-
tains more dihydrate relative to other samples with is predomi-
nantly consisting of the monohydrate form of calcium oxalate
present in Group I [33]. Calcium oxalate hydrates are characterized
by one (COM) or two (COD) DTA endothermic peaks of evolving
water. The carbon monoxide is oxidized into carbon dioxide, so
an exothermic peak appears in the second step. Third one shows
an endothermic peak. An example of each hydrate is shown in
Fig. 5. The TG (weight loss) and DTA (endothermic and exothermic
peaks) curves of COD and COM standards, displayed three typical
steps [34,35]

Fig. 1. FT-IR spectra of urinary stones (KS1–KS5).
In group II, KS10 stone samples XRD patterns of the samples
under investigation are analyzed, the ‘d’ values of samples are com-
pared with standard ‘d’ values from JCPDS data. The XRD pattern
(Fig. 4) of calcium oxalate monohydrate (COM), calcium phosphate
(Apatite), magnesium ammonium phosphate (MAPH) and uric acid
with either ‘d’ values and intensities of experimental values and
standard values are shown in Table 4 and the comparing the ‘d’
and I/Io values of Apatite and MAPH component are present in
the mixed stone of Calcium oxalate monohydrate (COM), Calcium
phosphate (Apatite), Magnesium ammonium phosphate (MAPH)
and Uric acid are presented in Table 3. The XRD spectra of KS10
are shown in Fig. 4.
TGA–DTA analysis of human urinary stone
Calcium oxalate hydrates
The three major steps of weight loss beginning first step at 95–
244 °C, second step 244–542 °C, third step 542–746 °C, correspond
to Calcium Oxalate decomposition. Relatively, identical region of
weight loss up to 800 °C calcium carbonate decomposition.
According to the TG curve, the first gradual weight loss with an
I Step
Peak endo bounded 2H2 O CaC2 O4  2H2 O ! CaC2 O3 þ 2H2 O
II Step
Peak exo CO CaC2 O4 ! CaCO3 þ CO
III Step
Peak endo CO2 CaCO3 ! CaO þ CO2
Magnesium ammonium phosphate hexahydrate (MAPH)
The first is attributed endothermic peak to the loss of water and
ammonia [35] the second one is due to magnesium pyrophosphate
crystallization of second step is characterized by a continuous
weight loss until about 800 °C [33]. The TG transition temperatures
first step are in between 113 and 477.5 °C, second step 573–685 °C
respectively, The TG data shows 33%, weight loss of 1st step and
weight loss of 1.1%, second step. The thermal analysis of urinary
stone yielded a curve that suggested MAPH as part of its constitu-
ent. The magnesium ammonium phosphate hexahydrate decom-
poses, during heating, to magnesium hydrogen phosphate and
then to magnesium pyrophosphate, as shown below reaction
[35]. The thermal curves, obtained under nitrogen flow, were
similar and showed an endothermic DTA peak at indicate loss of
1400 R. Selvaraju et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 1397–1402

Table 2
FT-IR spectral data of urinary stones (KS6–KS10).

Wavenumber (cm1) Tentative assignments Compounds

KS6 KS7 KS8 KS9 KS10
3431 3425 3486 3403 – OAH stretching COM
3340 – – – – OAH stretching COM
3257 3259 3243 3251 OAH stretching COM
3064 – – – – OAH stretching COM
2929 2933 2918 2918 2850 CAH stretching COM
1624 1618 1655 1649 1641 Vibration of C@O COM
1469 – – – – CO2
3 stretching Calcium carbonate
1435 1436 1436 1421 1438 PO4 group MAPH
1319 1316 – – 1315 Vibration of CAO COM
– – – – 1085 PO3
4 stretching Apatite
– – – – 1053 PO3
4 stretching Apatite
1035 1028 – 1028 1016 PO3
4 stretching Apatite
– – – – 979 CAO stretching COM
– – – – 904 C@C stretching COM
875 870 – 870 879 CAC stretching COM
777 777 – – – CAC stretching COM
– – 756 764 759 PO3
4 group MAPH
663 673 – – – Out-of-plane OAH bending COM
603 604 598 PO3
4 bending Apatite
569 567 574 567 567 PO3
4 bending Apatite
520 – – – – OACAO in plane bending COM
– – 468 468 462 NH+4 deformation Uric acid

Table 3
XRD spectral data of urinary stones (KS5).

Observed Standard hkl FWHM Minerals

0 0
I/Io I/Io
A)
d (Å d (Å
A)
7.150 71.5 7.570 100 020 0.1600 CaHPO42H2O
5.930 41.4 5.930 100 101 0.1600 CaC2O4
5.609 64.7 5.840 90 120 0.1200 CaC2O4
4.270 68.6 4.280 60 – 0.1200 CaC2O4
3.460 56.8 3.650 70 020 0.1600 CaC2O4H2O
2.819 100 2.8140 100 211 0.2400 Ca5(PO4)3 (OH)
2.749 73.2 2.720 60 300 0.0400 Ca5(PO4)3 (OH)

Fig. 2. FT-IR spectra of urinary stones (KS6–KS10).

Fig. 4. XRD spectrum of urinary stones (KS10).

Fig. 3. XRD spectrum of urinary stones (KS5).
 R. Selvaraju et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 1397–1402 1401

Table 4 water and ammonia 111 °C, a small exothermic due to magnesium
XRD spectral data of urinary stones (KS10). pyrophosphate. Terminal weight loss is observed to 34.4% is shown
Observed Standard hkl FWHM Minerals in Fig. 6.
0 0
I/Io I/Io
A)
d (Å d (Å
A)
6.165 54.0 6.142 61 010 0.0800 MgNH4PO46H2O
5.945 72.9 5.930 100 101 0.1600 CaC2O4
3.804 24.8 3.837 20 311 0.1600 C5H4N4O3
3.669 100.0 3.660 100 040 0.1600 CaC2O4
3.453 28.7 3.450 25 1010 0.2400 Ca3(PO4)2
2.989 56.4 2.920 60 321 0.0800 CaC2O4
2.799 41.2 2.802 35 040 0.2400 NH4MgPO46H2O
2.517 38.2 2.553 33 202 0.1200 CaHPO42H2O
2.381 40.6 2.370 40 710 0.0800 UA
1.992 67.9 1.996 62 123 0.0800 CaHPO42H2O
1.860 20.9 1.868 20 500 0.1200 CaC2O4

Fig. 5. Thermal analysis of urinary stones (KS5).

Fig. 6. Thermal analysis of urinary stones (KS10).

1402 R. Selvaraju et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 137 (2015) 1397–1402
There is no further weight loss could be observed above 850 °C.
The FT-IR and XRD results are well agreement with TGA and DTA
results.
Conclusion
The application of the techniques FT-IR, XRD and TG–DTA
enable to explain the detailed characterization of mineralogical,
chemical composition, dehydroxylation of urinary stones. Present
study indicated that the urinary stones were observed in their dif-
ferent chemical compositions. FT-IR analysis results well coincided
with XRD and TG–DTA analyses performed on the samples of inter-
est. Thermal analysis showed the characteristic peaks for dehydra-
tion and decomposition of calcium oxalate, struvite, uric acid and
calcium bilirubinate, calcium phosphate or hydroxyapatite up to
800 °C. Thus, the results of FT-IR are helpful in the identification
of various forms of minerals present in the used urinary stones.
References
[1] Z. Jing, W. GuoZeng, J. Ning, Y. JiaWei, G. Yan, Y. Fang, Urol. Res. 38 (2010) 111–
115.
[2] E.K. Girija, S. Narayana Kalkura, P.B. Sivaraman, Y. Yokogawa, J. Sci. Ind. Res. 66
(2007) 632–639.
[3] M. Modlin, S. Afr, Med. J. 58 (1980) 652–655.
[4] M.S. Ansari, N.P. Gupta, A.K. Hemal, P.N. Dogra, A. Seth, M. Aron, Int. J. Urol. 12
(2005) 12–16.
[5] A. Alaya, A. Nouri, M.F. Najjar, Clin. Chem. Lab. Med. 49 (2011) 243–248.
[6] A.C. Persaud, M.D. Stevenson, D.R. Mahon, N.C. Christopher, Pediatrics 124
(2009) 1088–1094.
[7] A.A. Al-Eisa, A. Al-Hunayyan, R. Gupta, Int. Urol. Nephrol. 33 (2002) 3–6.
[8] M. Straub, J. Gschwend, C. Zorn, Pediat. Nephrol. 25 (2010) 1239–1244.
[9] M. Gretchen, M. Neil (Eds.), Analysis of stones, Kidney stones: Medical and
Surgical Management, New York, 1996, pp. 323–335.
[10] F.S. Silvia, S.L. Silva, E.F. Daher, G.B. Silva Junior, R.M. Mota, C.A. Bruno da Silva,
Clin. Chem. Lab. Med. 47 (2009) 561–564.
[11] I. Dursun, H.M. Poyrazoglu, R. Dusunsel, Z. Gunduz, M.K. Gurgoze, D. Demirci,
Int. Urol. Nephrol. 40 (2008) 3–9.
[12] K.H. Safaa Khalil, A. Mohamad, J. Appl. Sci. Res. 3 (2007) 387–391.
[13] G. Sahubert, Urol. Res. 14 (2006) 1–5.
[14] K. Sarica, Urol. Res. 34 (2006) 96–101.
[15] U.S. Alon, Pediat. Nephrol. 24 (2009) 2129–2135.
[16] C.V. Raman, R.S. Krishnan, Nature 121 (1928) 501–502.
[17] S.D. Ruchita, Y.K. Agrawal, Vib. Spectros. 57 (2011) 163–176.
[18] P.P. Carmona, J. Bellanato, E. Escolar, Biospectroscopy 3 (1997) 331–346.
[19] M. Daudon, M.F. Protat, R.J. Reveillaud, H. Jaeschke, Kidney Int. 23 (1993) 842–
850.
[20] V.R. Kodati, G.E. Tomasi, J.L. Turumin, A.T. Tu, Appl. Spectros. 45 (1991) 581–
583.
[21] R. Selvaraju, G. Thiruppathi, A. Raja, Spectrochim. Acta A 93 (2012) 260–
265.
[22] R. Selvaraju, A. Raja, G. Thiruppathi, Spectrochim. Acta A 99 (2012) 205–210.
[23] S. Matsuzaki, K. Matsushita, K. Tanikawa, A. Masuda, F. Matsunaga, Int. J. Urol.
2 (1995) 235–237.
[24] E.V. Wilson, M.J. Bushiri, V.K. Vaidyan, Spectrochim. Acta A 77 (2010) 442–
445.
[25] R.L. Frost, Anal. Chim. Acta 517 (2004) 207–214.
[26] L. Estepa, M. Daudon, Biospectroscopy 3 (1997) 347–355.
[27] T.H. Shing Hsu, S.L. Yang, C.L. Cheng, W.T. Cheng, Urol. Res. 39 (2011) 165–170.
[28] L. Estepa, M. Daudon, Clin. Chim. Acta 298 (2000). 1–1.
[29] Y.H. Lee, W.C. Huang, J.Y. Tsai, J.K. Huang, J. Urol. 161 (1999) 1453–1457.
[30] G. Kanchana, P. Sundaramoorthi, G.P. Jeyanthi, J. Miner. Mater. Charact. Eng. 8
(2009) 161–170.
[31] J. Bellanto, Infrared spectroscopy of urinary calculi, in: J.E.A. Wickham, A.
Colinbuck (Eds.), Renal Tract Stone: Metabolic Basis and Clinical Practise,
Churchill Livingstone, Newyork, 1990, pp. 45–57.
[32] J. Kaloustian, F.Tarek. El-Moselhy, H. Portugal, Clin. Chim. Acta 334 (2003)
117–129.
[33] M. Berenyi, G. Liptay, J. Therm. Anal. 3 (1971) 437–443.
[34] B.D. Mitchell, A.C. Birnie, Biological materials, in: R.C. Mackenzie (Ed.),
Differential Thermal Analysis, vol. 1, Academic Press, London, 1970, pp. 673–
704.
[35] C.J. Keattch, D. Dollimore, An Introduction to Thermogravimetry, 2nd. ed.,
Heyden & Son Ltd., England, 1975.