Review

What is the State of the Stone Analysis

Techniques in Urolithiasis?
Abbas Basiri,1 Maryam Taheri,1 Fatemeh Taheri2

1Urology and Nephrology

Purpose: To compare and evaluate the mostly used methods of urinary stone anal-
Research Center, Shahid ysis.
Labbafinejad Medical
Center, Shahid Beheshti
University of Medical Sci- Materials and Methods: We searched PubMed and Google Scholar for “urolithi-
ences, Tehran, Iran
2Department of Clinical asis, nephrolithiasis, renal stone, and kidney stone” combined with “stone analysis,
Biochemistry, Faculty of
Medical Sciences, Tarbiat
spectroscopy, X-ray diffraction, chemical analysis, mass spectrometry, and laser-
Modares University, Teh- induced breakdown spectroscopy, review article, and quality control assessment.”
ran, Iran

Results::HLGHQWL¿HGDUWLFOHVDERXWUHYLHZVRIWKHSULQFLSOHVRIVWRQHDQDO\-
sis techniques and their quality control trials. Seven articles were not in English
language; hence, were omitted from this review. The remaining 17 articles and
their related references were studied thoroughly. There are various chemical and
physical techniques available for urinary stone analysis. The correct stone analysis
has to identify not only all stone components, but also the molecular structure and
Corresponding Author: crystalline forms of them with the exact quantitative determination of each com-
ponent.
Maryam Taheri, MD
Urology and Nephrology
Research Center, No.103, Conclusion: The knowledge of urinary stone composition is important for un-
9th Boustan St., Pasdaran
Ave., 1666677951,
derstanding pathophysiology, choice of treatment modality, and prevention of re-
Tehran, Iran FXUUHQFHVRIXUROLWKLDVLVEXWXSWRQRZDVWDQGDUGPHWKRGKDVQRWEHHQGH¿QHG
Although there are many techniques available for identifying the urinary stone
Tel: +98 21 2256 7222
Fax: +98 21 2256 7282 composition and structure, no single method can provide all the requiring informa-
E-mail: taheri233@yahoo. tion. Therefore, a combination of structural and morphological tests is needed for
com
this purpose.
Received April 2012
Accepted April 2012 Keywords: kidney calculi, chemical analysis, spectroscopy, X-ray diffraction

UROLOGY JOURNAL Vol. 9 | No. 2 | Spring 2012 | 445

INTRODUCTION mogravimetry, optic polarizing microscopy, scan-

T
he incidence of nephrolithiasis has consid- ning electron microscopy, and different methods
erably increased throughout the world in RIVSHFWURVFRS\$W¿UVWZHGHVFULEHWKHSULQFLSOHV
the last twenty years. The treatment of of these methods and then, compare their accuracy
urinary stone can be painful, stone removal often and practical application according to our literature
requires surgery, and renal failure occurs in about review.
3% of patients. Furthermore, the recurrence rates Wet Chemical Analysis
PD\EHDVKLJKDVWRSHU\HDUDQGPD\ Although wet chemical technique is the most
reach 50% within 5 years if a proper management, widely used approach for stone analysis in routine
stone analysis, and follow-up are not applied. laboratories, it can only identify the presence of
The most frequent component of urinary calculi is individual ions and radicals without differentiat-
calcium, which is the major constituent of nearly LQJDVSHFL¿FFRPSRXQGLQPDQ\VWRQHW\SHVDQG
75% of stones. Urinary stone is mostly composed mixtures. An external quality assurance scheme
of calcium oxalate about 60%, mixed calcium oxa- showed relatively poor performance of qualitative
ODWH DQG K\GUR[\ DSDWLWH  XULF DFLG DSSUR[L- and semi-quantitative wet chemical tests, includ-
mately 10%, struvite (magnesium ammonium ing commercial kits. However, its performance
SKRVSKDWH DERXW  EUXVKLWH  DQG F\VWLQH can improve by using quantitative wet chemical
1%. The purposes of stone analysis are qualita- approach, in which the same routine quantitative
tive differentiation of all stone components and chemical analysis methods for blood and urine are
their semiquantitative determination. The aim of used for a suitably prepared solution of the stone.
this review is to compare the principles and prac- Thermogravimetry
tical application of various chemical and physical Since the 1970s, thermogravimetric analysis (TG
techniques used for urinary stone analysis. RU7*$KDVEHHQH[WHQVLYHO\DSSOLHGIRUDQDO\]-
MATERIALS AND METHODS
According to our search on PubMed and Google
Scholar for “urolithiasis, nephrolithiasis, renal
stone, and kidney stone” combined with “stone
analysis, spectroscopy, X-ray diffraction, chemi-
cal analysis, mass spectrometry, and laser-induced
breakdown spectroscopy, review article, and qual-
ity control assessment.”
RESULTS
ing kidney stones. Thermogravimetry is a viable,
fast, and simple technique based on continuous re-
cording of both the temperature and weight loss of
the material during a progressive temperature in-
crease to 1000 ºC in an oxygen atmosphere.
$V HDFK VXEVWDQFH KDV LWV RZQ VSHFL¿F W\SH RI
transformation, the starting and ending tempera-
ture of transformation, the amount of change in
weight, and enthalpy, the nature of the substance
ZLOO EH FODUL¿HG E\ WKH SDWWHUQ RI ZHLJKW FKDQJH
and the magnitude of this change indicates the pro-
:HLGHQWL¿HGDUWLFOHVDERXWUHYLHZVRIWKHSULQ-
ciples of stone analysis techniques and their quality
control trials. Seven articles were not in English
language; hence, were omitted from this review.
The remaining 17 articles and their related refer-
ences were studied thoroughly.
Currently, the following methods are available
portion.
Optic Polarizing Microscopy
The base of this technique is the interaction of po-
larized light with crystals of stones. After the stone
is fractured and the material is removed from vari-
ous points of it, it can be assessed under the polar-
izing microscope using a drop of the appropriate
for stone analysis: Wet chemical analysis, ther- refractive index liquid. Parameters which iden-

446 | Review
 Stone Analysis Techniques in Urolithiasis | Basiri et al

tify the stone minerals include the color, refraction however, it has become as a popular reliable meth-
of light, and double refraction. od for in-vitro quantitative stone analysis in the last
Scanning Electron Microscopy (SEM) decade.,QIUDUHGVSHFWURVFRS\LVDVSHFL¿FUDS-
Scanning electron microscopy is a precise tech- id, and versatile method, which uses IR radiation in
nique for the study of morphology of urinary cal- order to cause atomic vibrations, consequently, en-
culi. This technique is non-destructive and reveals HUJ\DEVRUSWLRQDQG¿QDOO\DSSHDUDQFHRIDEVRUS-
details about stones 1 to 5 nm in size, without tion bands in the IR spectrum of stone samples.
FKDQJLQJ WKH VSHFL¿F PRUSKRORJ\ RI WKH FRPSR- Two different IR spectroscopy approaches are
nents. Furthermore, it can produce very high-res-
olution images of a sample surface.
Spectroscopy
Spectroscopy is the study of the interaction be-
tween matter and radiated energy. Spectroscopy
common: The direct IR transmission, in which, the
stone material is mixed with potassium bromide
and compressed to form a disc, which is used for
the analysis. Therefore, stone material cannot be
recovered for further supporting analysis, such as
WHFKQLTXHVFDQEHFODVVL¿HGLQVHYHUDOZD\VZKLFK wet chemical tests. But in non-destructive ap-
are summarized in Table 1.Here the principles proach, such as photo-acoustic detection,recov-
and practical application of mostly used methods ery of the sample is possible.
of spectroscopy will be presented. A more recent technique in IR spectroscopy is the
Infrared (IR) Spectroscopy PHWKRG RI DWWHQXDWHG WRWDO UHÀHFWLRQ WHFKQLTXH
,QIUDUHGVSHFWURVFRS\ZDVXVHG¿UVWO\LQ $75ZKLFKLVDSSOLFDEOHIRUVRIWVDPSOHVFur-
Table 1. Classification of the spectroscopy techniques.
1. Type of radiated energy

Electromagnetic radia- Classified by the wavelength region of the spectrum and includes micro-
tion wave, terahertz, infrared, near infrared, visible and ultraviolet, x-ray, and
gamma spectroscopy.
Particles Electrons and neutrons can also be a source of radiative energy
Acoustic spectroscopy Involves radiated pressure waves
2. Nature of the interaction

Absorption
Emission
Elastic scattering and Determines how incident radiation is reflected or scattered by a material,
Reflection spectros- such as X-xay Diffraction
copy
Impedance spectros-
copy
Inelastic scattering Raman scattering
Coherent or resonance Nuclear magnetic resonance (NMR) spectroscopy
spectroscopy
3. Type of material

Atoms Atomic absorption spectroscopy (AAS)

Atomic emission spectroscopy (AES)
Flame emission spectroscopy
Inductively coupled plasma atomic emission spectroscopy
X-ray spectroscopy
X-ray fluorescence (XRF)
Molecules Infrared and Raman spectroscopy
Crystals
Nuclei NMR spectroscopy

UROLOGY JOURNAL Vol. 9 | No. 2 | Spring 2012 | 447

thermore, sample preparation for this technique is
very easy, as it does not require mixing the sam-
ple with an IR inactive material, such as potassium
bromide, prior to analysis.
X-Ray Powder Diffraction (XRD)
X-ray powder diffraction uses monochromatic
X-rays for identifying the constituents of a renal
stone based on the unique diffraction patterns pro-
duced by a crystalline material. Crystal moieties of
DVWUXFWXUHGLIIUDFWRUUHÀHFWWKHSHQHWUDWHG;UD\V
lyze small amounts of stones, and the rapidity of
polarizing microscopy are the advantages of this
technique over chemical analysis. Silva and col-
leagues studied a sample of 50 stones retrieved
from patients in Brazil in order to compare the
chemical with morphological kidney stone com-
position analysis. They found that unlike morpho-
logical analysis, chemical analysis can only detects
calcium and oxalate separately without differenti-
ating the crystalline types. Identifying the crystal-
in particular patterns. line form is very useful for planning therapy, eg,
Elementary Distribution Analysis (EDAX) FDOFLXP R[DODWH GHK\GUDWH &2' LV DVVRFLDWHG
(OHPHQWDU\GLVWULEXWLRQDQDO\VLV('$;LVXVHG with hypercalciuria while calcium oxalate mono-
to obtain the percentage of composition of all stone K\GUDWH&20LVPRUHFORVHO\UHODWHGWRK\SHUR[-
VDPSOHHOHPHQWVDQGLVVLJQL¿FDQWO\XVHIXOIRUUHF- aluria. Therefore, they offered using both types of
RJQL]LQJXQNQRZQFU\VWDOVQRWLGHQWL¿HGE\RUGL- analysis routinely for a better understanding of the
nary light microscopy or SEM alone. Elementary mechanisms involved in lithogenesis.
GLVWULEXWLRQ DQDO\VLV FDQ EH XVHG WR FRQ¿UP WKH Jhaumeer-Laulloo and Subratty employed wet
SEM results and also evaluate the percentage of chemical tests and IR spectroscopy techniques to
the different elements present in a sample. DQDO\]HNLGQH\VWRQHVIURPSDWLHQWV7KHLU¿QG-
ings revealed that the chemical analysis method
DISCUSSION ZDV QRW DFFXUDWH DQG FRXOG UHVXOW LQ VLJQL¿FDQW
Nephrolithiasis is a recurrent condition with con- clinical errors. They also showed that the spec-
siderable morbidity. While in symptomatic stone troscopic methods were applicable for smaller
episodes the appropriate treatment is necessary, amount of sample and were able to identify the dif-
prophylactic workup to prevent recurrences is also ferent constituents of the renal stones.
of great importance, which is possible by a com- Singh analyzed urinary stones of 50 patients by
plete metabolic workup and a suitable stone analy- )RXULHU7UDQVIRUP,5)7,5DQGRIIHUHGWKDW)7
sis. ,5LVDQHI¿FLHQWDQGDFFXUDWHWHFKQLTXHWRGHWHU-
The chemical composition of urinary calculi was mine urinary stone composition. He also mentioned
¿UVWO\UHSRUWHGDWWKHHQGRIWKHth century, when that using computerized IR spectrophotometer and
important chemical components of urinary calculi, large reference library enable us to determine ex-
VXFKDVXULFDFLG6FKHHOHDQGF\VWLQH:RO- act quantitative stone composition, and this meth-
ODVWRQZHUHGLVFRYHUHG$IWHUWKHV\VWHPDWLF od should be extended to all urolithiasis centers.

VWXGLHVE\+HOOHUDQG8OW]PDQQWKH While Estepa and coworkers depicted the pos-
chemical analysis of urinary calculi was presented VLELOLW\RIIDOVHLGHQWL¿FDWLRQGXULQJXVLQJ,5EH-
as an established routine. cause of library incompleteness and also consider-
,Q  3ULHQ DQG )URQGHO LQWURGXFHG WKH SRODU- able differences between the spectra of natural and
izing microscopy as an analytical tool for identi- synthetic compounds. For example, they observed
¿FDWLRQ RI WKH FU\VWDOOLQH FRQVWLWXHQWV RI FDOFXOL that the spectrum of human whewellite shows a
by refractive index measurements. The ability peak at 1315 cm–1 while the peak is at 1319 cm–1
WRLGHQWLI\VSHFL¿FPLQHUDOVWKHFDSDELOLW\WRDQD- for the synthetic one. On the other hand, a peak at

448 | Review
 Stone Analysis Techniques in Urolithiasis | Basiri et al

1319 cm–1 in human stone samples corresponds to cording to these results, Rebentisch offered that
a mixture of whewellite and weddellite in 50/50 X-ray diffraction and IR spectroscopy methods
proportions. 7KHUHIRUH IDOVH LGHQWL¿FDWLRQ RI give comparable and highly acceptable analytical
weddellite may occur using the search procedure UHVXOWVDQGFDQEHFODVVL¿HGDVUHIHUHQFHPHWKRGV
if synthetic whewellite is included as a reference for the analysis of urinary calculi.The fourth In-
in libraries. ternational Ring Test for checking the quality of
5RVHDQG:RRG¿QHXVHG7*$IRUWKHDQDO\VLVRI methods for urinary calculus analysis, conducted
stones and supported its ability to produce fast by Rebentisch and colleagues in 1988, demon-
and quantitative results.However, limitations of strated that the method of XRD is clearly superior
TGA require relatively large amount of material
for optimal resolution and non-recovery of sample.
Furthermore, similarity in ignition temperatures
and rates of disintegration of some closely related
compounds, such as purines, may make the identi-
to IR spectroscopy. Also in external quality as-
sessment of analysis of urinary calculi, which was
commenced in 1991, they suggested that the use of
chemical methods should be discontinued because
RILWVXQDFFHSWDEOHTXDOLW\LQDSSUR[LPDWHO\
¿FDWLRQGLI¿FXOW$VFDOFLXPSKRVSKDWHDSDUWIURP of laboratories.
WKH EUXVKLWH IRUP &D+32+2 VLOLFD DQG Hesse and associates designed a twice-yearly ring
calcium pyrophosphate display very little weight trials quality control survey to examine the quality
change on heating, TGA cannot convincingly iden- of urinary stone analysis based on synthetic prod-
tify them from each other. ucts in averagely 100 laboratories since 1980. The
Since 1970s, physico-chemical techniques have methods employed for these analyses were based
been increasingly employed for urinary stone anal- on chemical analysis, IR spectroscopy, and X-ray
ysis, which resulted in discovering numerous crys- GLIIUDFWLRQ 7KH UHVXOWV RI  ULQJ WULDOV  WR
talline elements in urinary stones. Many clini-  LOOXVWUDWHG WKDW DW ¿UVW WKH DQDO\VHV ZHUH
cal laboratories employed X-ray diffraction and carried out using chemical methods for more than
IR spectroscopy as reference techniques for stone  RI WKH SDUWLFLSDQWV7KLV ¿JXUH FRQVLGHUDEO\
analysis. Thereafter, a lot of studies were designed GHFUHDVHGWRLQ2QWKHFRQWUDU\WKHXVH
to compare the quality of these methods in ad- of IR spectroscopy progressively increased to 79%.
dition to quality control surveys that were conduct- The number of specialized laboratories which used
ed for improving the standards of them, some of
which are presented as follows.
Rebentisch presented the result of six external
TXDOLW\DVVHVVPHQWVXUYH\VRIDYHUDJHO\ODER-
UDWRULHV IURP  FRXQWULHV WKDW XVHG VL[ TXDQWLWD-
tive and various qualitative analytical techniques
X-Ray diffraction was constantly about 5% to 9%.
Additionally, these ring trials revealed that error
rates for IR spectroscopy and X-ray diffraction
were only limited to individual substances, where-
as for the chemical methods very high proportion
of errors occurred with both the pure substances
during 1983 and 1988. He used both standard of DQGELQDU\PL[WXUHVWR7KHUHIRUHWKH
quality and the mean deviation as two determining majority of laboratories stopped using chemical
parameters for ranking the methods, which were analysis, which is now considered to be obsolete.

from the best performance to worst in the follow-
ing order: X-ray diffraction, IR spectroscopy, ul- Kasidas and associates analyzed the results of ex-
WUDPLFURFKHPLFDO DQDO\VLV 80&$ SRODUL]DWLRQ ternal quality assurance for urinary stone analysis
PLFURVFRS\0,GLIIHUHQWLDOWKHUPRDQDO\VLV7$ LQ*UHDW%ULWDLQGXULQJDQG$FFRUGLQJ
DQGTXDQWLWDWLYHFKHPLFDODQDO\VLV47&$ Ac- WRWKHLU¿QGLQJVPRVWRIWKHSDUWLFLSDWLQJODERUD-

UROLOGY JOURNAL Vol. 9 | No. 2 | Spring 2012 | 449

WRULHVXVHGZHWFKHPLFDODQDO\VLVZLWKRQO\ visualize even small struvite regions in stones.
55% to 65% accuracy versus IR with 85% to 90% Siritapetawee and Pattanasiriwisawa used X-ray
correct analyses. Another study in china con- DEVRUSWLRQQHDUHGJHVSHFWURVFRS\;$1(6DQG
¿UPHG WKH UHOLDELOLW\ DQG DFFXUDF\ RI ;UD\ GLI- XRPD for analyzing 15 human urinary stones.
fraction for qualitative and quantitative analysis of Comparing the result of XANES spectra of un-
urinary stones. known compounds from human kidney stones with
7KHEHQH¿WVDQGGLVDGYDQWDJHVRIWKHPRVWFRP- the diffractogram data of the XRPD, it was shown
mon techniques based on the reviewed studies are that these two techniques agreed well with each
VXPPDUL]HGLQ7DEOH other, while XANES required a smaller amount of
In addition to the most common techniques that each sample than XRPD for analysis.
were mentioned above, there are other techniques In 1995, the use of nonenhanced computed tomog-
which may have in-vivo application in practice UDSK\ &7 LPDJLQJ WR LGHQWLI\ XUROLWKLDVLV ZDV
RU UHYHDO PRUH GHWDLOHG LQIRUPDWLRQ RQ ¿QH PR- ¿UVWUHSRUWHGE\6PLWKDQGDVVRFLDWHVDQGQRZLW
lecular structure or several unexpected trace ele- has become the standard diagnostic tool for evalu-
ments mainly in the nuclear region of the stones. ation of patients with renal colic. Due to the
Meanwhile, often these techniques are not useful
in routine laboratory for being costly and requiring
special expertise or sample preparation.
Kim and colleagues analyzed 86 consecutive uri-
nary stones by X-ray analysis and compared the
¿QGLQJVZLWKWKRVHRIWKUHHRWKHUPHWKRGV;UD\
diffraction, IR spectrometry, and chemical analy-
sis. This study indicated that the sensitivity of X-
ray analysis was several times more than other
three methods, especially in detection of apatite.
HPHUJHQFH RI QRQFRQWUDVW &7 1&&7 LQ WKH DV-
VHVVPHQW RI DFXWH ÀDQN SDLQ DQG WKH VXEVHTXHQW
DYDLODELOLW\RIWKHDWWHQXDWLRQFRHI¿FLHQWPHDVXUH-
ment, several groups were interested in comparing
attenuation and stone composition in vitro. Studies
involving the use of single-energy CT technology
have shown that some information about stone
composition may be gained, enabling differentia-
tion between uric acid and calcium stones on the
basis of their different attenuations, with lower at-
This study also offered X-ray analysis as a particu- WHQXDWLRQYDOXHVLQ+RXQV¿HOGXQLWVIRUXULFDFLG
larly suitable method for detection of rare inorgan- stones.7KHGLI¿FXOW\LQWKHVHVWXGLHVZDVUHODWHG
ic components of urinary stones, such as silica and to the considerable overlap of attenuation values,
gypsum. ZKLFKSUHYHQWHGKLJKOHYHOVRIVSHFL¿FLW\
Batchelar and coworkers revealed that the X-ray Dual-energy CT by low and high-energy scanning
coherent scatter analysis is a novel technique for is capable to differentiate various materials with
intact stone analysis using monoenergetic X-ray similar electron densities, but different photon ab-
from the standard diagnostic X-ray equipment. sorption. Therefore, it may not only contribute
Because the coherent scatter properties is related to
the molecular structure of the scattering media of
each of the stone components, COM, cystine, mag-
nesium ammonium phosphate, and calcium phos-
phate showed a distinct coherent scatter pattern,
which matches that of a pure chemical sample.
Wignall and associates suggested that coherent X-
ray scatter would be useful in future studies of the
ability of commercial laboratories because it can
450 | Review
WRWKHLGHQWL¿FDWLRQEXWDOVRWRWKHFKHPLFDOFKDU-
acterization of stones in the urinary tract, which
could be useful in surgical or medical treatment
decisions. Hidas and coworkers used dual-energy
CT to pre-operatively assess the composition of
XULQDU\VWRQHVLQSDWLHQWVDQGFRPSDUHGWKHUH-
sults with postoperative in-vitro X-ray diffraction
analysis. They found that dual-energy CT was able
to characterize the kidney stone composition with
 Stone Analysis Techniques in Urolithiasis | Basiri et al

DFFXUDF\%XWWKLVWHFKQLTXHFRXOGQRWLGHQ- transported to the plasma often by argon as a car-

tify struvite stones which had attenuation ratios rier gas. The plasma causes the material to be ion-
WKDWRYHUODSSHGZLWKFDOFL¿HGVWRQHUDWLRVDQGWKXV ized and moved through to the mass spectrometer,
could not be assessed reliably. which selectively detects ions at a given mass-to-
By combination of X-ray attenuation values and charge ratio.
morphological appearance, micro CT can identify Kontoyannis and colleagues analyzed mineral
¿QHVWRQHVWUXFWXUHDQGLWVPLQHUDOFRPSRQHQW components of a urinary stone forming layers with
especially for apatite. Furthermore, detection the use of three spectroscopic methods: Raman
of the following 6 minerals, uric acid, COM, COD, VSHFWURVFRS\56)7,5DQG;5'5DPDQVSHF-
cystine, struvite, and hydroxy apatite, in pure or troscopy could analyze the various mineral layers
heterogeneous urinary stones is possible without in the kidney stones by focusing the laser beam
having overlapped ranges of micro attenuation at the desired layer, whereas application of FT-IR
values. However, the study by Krambeck and produced overlapping broad bands and XRD could
DVVRFLDWHVUHYHDOHGWKDWWKHLGHQWL¿FDWLRQRIPLQHU- not analyze the mineral components of the various
als with lower X-ray attenuation, such as struvite, layers of small stones precisely since the material
is not always possible as a minor inclusion in par- FRQWDLQHGLQWKHVHVWRQHVZDVQRWVXI¿FLHQW
ticular mineral. In addition, its use in routine labo- In the study by Cytron and associates, analysis of
ratory has not been exploited because it costs 10 chemical composition of the stones and their con-
times more than FT-IR. centrations were determined by analysis of the
/DVHULQGXFHGEUHDNGRZQVSHFWURVFRS\/,%6LV XULQHVDPSOHVIURPSDWLHQWVEDVHGRQ¿EHURSWLF
a simple, rapid, and remote technique, which per- IR spectroscopy. The urine samples were collect-
PLWVUHDOWLPHTXDOLWDWLYHDQGTXDQWLWDWLYHLGHQWL¿- HGZLWKLQKRXUVDIWHUH[WUDFRUSRUHDOVKRFNZDYH
cation of the major and trace elements present in the OLWKRWULSV\7KLVVWXG\RIIHUHGWKDW¿EHURSWLF,5FDQ
calculus. In this technique, a pulsed laser beam be applied as an alternative method for complete
is focused on the surface of the sample in order and quick metabolic evaluation of patients without
to produce high-density plasma that excites vari-
ous atomic elements and elemental transitions in
the focal volume. Singh and associates performed
LIBS to estimate the quantitative elemental con-
stituents distributed in different parts of the kidney
stones obtained directly from 5 patients by surgery,
and compared the results with that of inductively
sample preparation.
According to the studies described above, there
are many different methods available for urinary
stone analysis, but the fact is that no single method
LVVXI¿FLHQWWRSURYLGHDOOWKHFOLQLFDOO\XVHIXOLQ-
formation about the structure and composition of
the stones. Therefore, a combination of these tech-
FRXSOHG SODVPD PDVV VSHFWURPHWU\ ,&306 niques is advised. Fazil Marickar and coworkers
Both LIBS spectra and ICP-MS analysis showed revealed that the combination of optical micros-
that the major constituent of the kidney stones was copy and IR spectroscopy of core, cross section,
calcium. and surface of calculi is an accurate and reliable
Laser ablation inductively coupled plasma mass PHWKRGIRULGHQWL¿FDWLRQRIWKHVWRQHVWUXFWXUHLWV
VSHFWURPHWU\ /$ ,&306 LV D QRYHO WHFKQLTXH FU\VWDOOLQHFRPSRVLWLRQDQGTXDQWL¿FDWLRQRIVWRQH
for the study of biological and geological sam- components while being highly cost-effective.
ples, which uses a focused laser beam issued Uldall showed that combination of X-ray diffrac-
to mobilize sample material as droplets or vapor tion or IR spectrometry and wet chemistry may be
from the sample surface. Thereafter, the material is suitable as reference methodology.

UROLOGY JOURNAL Vol. 9 | No. 2 | Spring 2012 | 451

In the literature, other combinations of methods, the calculus are analyzed separately.
such as TGA with X-ray diffraction by Konjiki and
colleagues, and TG with FT-IR spectroscopy by CONCLUSION
Materazzi and associates, have been mentioned.
Also, Fazil Marickar and coworkers analyzed 10
mixed stones using FT-IR spectroscopy and SEM-
EDAX combination in order to get a thorough
understanding of mixed stone morphology, and
concluded that although FT-IR analysis is more
modern, less time-consuming, and more precise,
combination of SEM-EDAX will give a clear in-
dication of the structure of the stone on the surface
DQG LQWHULRU LQ DGGLWLRQ WR H[DFW TXDQWL¿FDWLRQ RI
In this review, we observed that although wet
chemical analysis technique of urinary stone is the
traditional gold standard, it has been replaced with
more accurate spectroscopy techniques, such as
FT-IR, XRD, CT scan, etc. Also we believe that
our current results provide compelling evidence
to support the notion that in addition to applying
combination techniques, analysis of different parts
of stone separately is of utmost importance.
all stone elements. More importantly, the study CONFLICT OF INTEREST
by Schubert revealed that useful results of any of None declared.
these methods are obtained when different areas of
Table 2. Comparison of the stone analysis techniques.
Test Advantages Disadvantages
Chemical t
&BTZ t
5JNFDPOTVNJOH
analysis t
MPX
DPTU t
3FRVJSJOH
MBSHF
TUPOF
TBNQMFT
IFODF

VOTVJUBCMF
GPS
UJOZ
TUPOFT
t
0OMZ
JEFOUJöDBUJPO
PG
UIF
QSFTFODF
PG
JOEJWJEVBM
JPOT
BOE
SBEJDBMT

rather than a specific compound, eg, unable to distinguish between
the two commonly occurring calcium stones (monohydrate/dihydrate)
Polarization t
$PTUFóDJFOU t)JHI
TVCKFDUJWF
FYQFSJFODF
JT
OFDFTTBSZ
microscopy t
2VJDL
FYBNJOBUJPOT t%JòFSFOUJBUJPO
PG
DPNQPOFOUT
JT
EJóDVMU
JO
TPNF
DBTFT
JO
UIF
HSPVQT

t
1PTTJCJMJUZ
UP
BOBMZ[F
WFSZ
TNBMM
TBNQMFT of uric acid, purine derivates, and calcium phosphates
t
"CJMJUZ
UP
EFUFDU
WFSZ
TNBMM
DPOUFOUT
PG
DPN-
ponents in the stone*
Infrared t
.PEFSBUF
DPTU t
5JNFDPOTVNJOH
QSFBOBMZUJD
QSFQBSBUJPO

spectros- t
2VJDL
FYBNJOBUJPO
t
5IF
QSPQFS
IPNPHFOJ[BUJPO
PG
UIF
TBNQMF
XJUI
QPUBTTJVN
CSPNJEF

copy t
1PTTJCJMJUZ
UP
FYBNJOF
TNBMM
TBNQMFT may affect the Infrared spectroscopy spectrum quality
t
&BTZ
QSFQBSBUJPO
XIFO
VTJOH
"UUFOVBUFE
5PUBM
t
3FTPMVUJPO
PG
UIF
BQQBSBUVT
BOE
SFQSPEVDJCJMJUZ
PG
UIF
TQFDUSVN

Reflection technique bands may affect its reliability
t
4FNJBVUPNBUJD
FWBMVBUJPOT
BSF
QPTTJCMF
BQQMZ- t
%JóDVMUZ
JO
EFUFDUJPO
PG
TNBMM
BNPVOUT
PG
DPNQPOFOUT
JO
TPNF
DBTFT


ing search–match functions eg, whewellite in weddellite or reverse, or urates and uric acid dihy-
t
6TFGVM
GPS
JEFOUJöDBUJPO
PG
PSHBOJD
DPNQP- drate in uric acid
nents or noncrystalline substances, eg, purines, t
%JóDVMUZ
JO
EFUFDUJPO
PG
TPNF
DPNQPOFOUT
EVP
UP
PWFSMBQQJOH
PG

proteins, or fat and drug metabolites their absorption band, such as carbonate in struvite stones or cystine
in whewellite or uric acid stone
X-ray dif- t
&BTZ
QSFQBSBUJPO t
)JHI
DPTU
fraction t
"VUPNBUJD
NFBTVSFNFOU t
6OBCMF
UP
EFUFDU
OPODSZTUBMMJOF
PS
BNPSQIPVT
TVCTUBODFT
t
2VBOUJUBUJWF
BOBMZTJT
t
&YBDU
EJòFSFOUJBUJPO
PG
BMM
DSZTUBMMJOF
DPNQP-
nents is possible
Thermo- t
'BTU t
3FRVJSJOH
SFMBUJWFMZ
MBSHF
BNPVOU
PG
TBNQMF
gravimetry t
4JNQMF t
/POSFDPWFSZ
PG
TBNQMF
t
%JóDVMUZ
JO
JEFOUJGZJOH
DMPTFMZ
SFMBUFE
DPNQPVOET

FH

QVSJOFT
Scanning t
/POEFTUSVDUJWF t
)JHI
DPTU
electron t
1PTTJCJMJUZ
UP
WJTVBMJ[F
UIF
DPNQPOFOUT
XJUIPVU

microscopy altering their spatial orientation and specific
morphology
* Whereas Kasidas and colleagues in another study showed that the Polarization microscopy cannot identify small amounts of crystal-
line material in mixtures.

452 | Review
 Stone Analysis Techniques in Urolithiasis | Basiri et al

REFERENCES 15. Gould N, Hallson PC, Kasidas GP, Samuell CT, Weir TB. Rapid
computer-assisted infrared analysis of urinary calculi using
1. Stamatelou KK, Francis ME, Jones CA, Nyberg LM, Curhan QIPUPBDPVTUJD
EFUFDUJPO
6SPM
3FT

GC. Time trends in reported prevalence of kidney stones in
16. Fazil Marickar YM, Lekshmi PR, Varma L, Koshy P. Elemen-
UIF
6OJUFE
4UBUFT

,JEOFZ
*OU

tal distribution analysis of urinary crystals. Urol Res.
2. Kasidas GP, Samuell CT, Weir TB. Renal stone analysis: why 
BOE
IPX
"OO
$MJO
#JPDIFN

17. Hesse A, Sanders G. Atlas of infrared spectra for the analysis
3. Estepa L, Daudon M. Contribution of Fourier transform in- of urinary concrements
4UVUUHBSU
(FPSH
5IJFNF
7FSMBH

frared spectroscopy to the identification of urinary stones 
BOE
LJEOFZ
DSZTUBM
EFQPTJUT
#JPTQFDUSPTDPQZ

 1SJFO
&-
'SPOEFM
$
4UVEJFT
JO
VSPMJUIJBTJT
UIF
DPNQPTJUJPO

69.
PG
VSJOBSZ
DBMDVMJ
+
6SPM

4. Krepinsky J, Ingram AJ, Churchill DN. Metabolic investiga-
19. Silva SFR, Matos DC, Silva SL, Daher EDF, Campos HH,
tion of recurrent nephrolithiasis: compliance with recom-
Silva CAB. Chemical and morphological analysis of kidney
NFOEBUJPOT
6SPMPHZ

stones: a double-blind comparative study. Acta Cirurgica
5. Pearle MS, Lotan Y. Urinary Lithiasis: Etiology, Epidemiol- #SBTJMFJSB

ogy, and Pathogenesis. In: Wein AJ, Kavoussi LR, Novick AC,
20. Jhaumeer-Laulloo S, Subratty A. Analysis of urinary calculi
Partin AW, Peters CA, eds. Campbell-Walsh Urology. Vol 1. 10
in Mauritius. Science & Technology-Research Journal.
FE
1IJMBEFMQIJB
4BVOEFST
&MTFWJFS


6. 4DIVCFSU
(
4UPOF
BOBMZTJT
6SPM
3FT

21. Estepa LM, Levillain P, Lacour B, Daudon M. Infrared analysis
7. Samuell CT, Weir TB, Hallson PC, Kasidas GP. An external of urinary stones: a trial of automated identification. Clin
quality assurance scheme for renal calculi: some early find- $IFN
-BC
.FE

ings In: Martin SM, Halloran SP, Green JE, eds. Proceedings
22. Rose GA, Woodfine C. The thermogravimetric analysis of
of the ACB National Meeting, Brighton, 9–13 May 1994.
SFOBM
TUPOFT
JO
DMJOJDBM
QSBDUJDF

#S
+
6SPM

-POEPO
"TTPDJBUJPO
PG
$MJOJDBM
#JPDIFNJTUT

23. Smith CL. Renal stone analysis: is there any clinical value?
 Lee HP, Leong D, Heng CT. Characterization of kidney
$VSS
0QJO
/FQISPM
)ZQFSUFOT

stones using thermogravimetric analysis with electron
EJTQFSTJWF
TQFDUSPTDPQZ
6SPM
3FT
 24. Hesse A, Schneider HJ, Hienzsch E. [Infra-red spectroscopy
PG
VSJOBSZ
DBMDVMJ>
%UTDI
.FE
8PDIFOTDIS

9. Douglas DE, Tonks DB. The qualitative analysis of renal
701.
calculi with the polarising microscope. Clin Biochem.
 25. Schneider HJ, Berenyi M, Hesse A, Tscharnke J. Compara-
UJWF
VSJOBSZ
TUPOF
BOBMZTFT
2VBOUJUBUJWF
DIFNJDBM
YSBZ

10. Harada Y, Tomita T, Kokubo Y, Daimon H, Ino S. Develop-
diffraction, infrared spectroscopy and thermo-analytical
ment of an ultrahigh vacuum high resolution scanning
QSPDFEVSFT
*OU
6SPM
/FQISPM

transmission electron microscope. J of Electron Microsc.
 26. )FTTF
"
,SVTF
3
(FJMFOLFVTFS
8+
4DINJEU
.
2VBMJUZ

control in urinary stone analysis: results of 44 ring trials
11. Crouch SR, Stanley R, Skoog, Douglas A. Principles of Instru-


$MJO
$IFN
-BC
.FE

mental Analysis

FE
#FMNPOU
$"
5IPNTPO
#SPPLT$PMF

2007. 27. Rebentisch G. Results of external quality assessment of
analysis of urinary calculi by various methods. Scand J Clin
12. Solli D, Chou J, Jalali B. Amplified wavelength–time trans-
-BC
*OWFTU
4VQQM

formation for real-time spectroscopy. Nature Photonics.
  Rebentisch G, Berg W, Pirlich W, Hommann D. Assessment
and maintenance of the quality of urolith analyses in a
13. Beischer DE. Analysis of renal calculi by infrared spectros-
comparison of methods. 4th International Ring Test to
DPQZ
+
6SPM

DIFDL
RVBMJUZ
*OU
6SPM
/FQISPM

14. Singh I. Renal geology (quantitative renal stone analysis) by
29. Rebentisch G, Muche J, Reinauer H. External quality assess-
'Fourier transform infrared spectroscopy'. Int Urol Nephrol.
ment of analysis of urinary calculi--a new scheme based

mainly on natural concrement materials. Scand J Clin Lab
*OWFTU
4VQQM


UROLOGY JOURNAL Vol. 9 | No. 2 | Spring 2012 | 453

30. Ouyang JM. [The application of X-ray diffraction in the
study of urinary stones]. Guang Pu Xue Yu Guang Pu Fen Xi.

31. Fazil Marickar YM, Lekshmi PR, Varma L, Koshy P. Problem in
analyzing cystine stones using FTIR spectroscopy. Urol Res.

32. Kim KM, Alpaugh HB, Johnson FB. X-ray microanalysis of
urinary stones, a comparison with other methods. Scan
&MFDUSPO
.JDSPTD

33. Batchelar DL, Chun SS, Wollin TA, et al. Predicting urinary
stone composition using X-ray coherent scatter: a novel
technique with potential clinical applications. J Urol.

34. Wignall GR, Cunningham IA, Denstedt JD. Coherent scatter
computed tomography for structural and compositional
stone analysis: a prospective comparison with infrared
TQFDUSPTDPQZ
+
&OEPVSPM

35. Siritapetawee J, Pattanasiriwisawa W. An attempt at kidney
stone analysis with the application of synchrotron radia-
UJPO
+
4ZODISPUSPO
3BEJBU

36. Gerber GS, Brendler CB. Evaluation of the urologic patient:
History, physical examination, and urinalysis. In: Wein AJ,
Kavoussi LR, Novick AC, Partin AW, Peters CA, eds. Camp-
bell-Walsh Urology. Vol 1. 10 ed. Philadelphia: Saunders
&MTFWJFS

37. Hidas G, Eliahou R, Duvdevani M, et al. Determination
of renal stone composition with dual-energy CT: in vivo
analysis and comparison with x-ray diffraction. Radiology.

 Bellin MF, Renard-Penna R, Conort P, et al. Helical CT evalu-
ation of the chemical composition of urinary tract calculi
with a discriminant analysis of CT-attenuation values and
EFOTJUZ
&VS
3BEJPM

39. Gottlieb RH, La TC, Erturk EN, et al. CT in detecting urinary
tract calculi: influence on patient imaging and clinical
PVUDPNFT
3BEJPMPHZ

40. Mostafavi MR, Ernst RD, Saltzman B. Accurate determina-
tion of chemical composition of urinary calculi by spiral
DPNQVUFSJ[FE
UPNPHSBQIZ
+
6SPM

41. Zarse CA, McAteer JA, Sommer AJ, et al. Nondestructive
analysis of urinary calculi using micro computed tomogra-
QIZ
#.$
6SPM


4VEP
5

,PIZBNB
/
.JOFSBMPHJDBM
OPUFT
PG
BQBUJUF

42. Matlaga BR, Williams JCJ, Evan AP. Calcium oxalate stones are
frequently found attached to Randall’s plaque. In: Evan AP,
Lingeman JE, Williams JCJ, eds. Renal stone disease. Proceed-
ings of the First International Urolithiasis Research Symposium.
.FMWJMMF
"NFSJDBO
*OTUJUVUF
PG
1IZTJDT
o

454 | Review
43. Pramanik R, Asplin JR, Jackson ME, Williams JC, Jr. Protein
content of human apatite and brushite kidney stones:
significant correlation with morphologic measures. Urol
3FT

44. Williams JC, Jr., Zarse CA, Jackson ME, Witzmann FA, McA-
teer JA. Variability of protein content in calcium oxalate
NPOPIZESBUF
TUPOFT
+
&OEPVSPM

45. Miller NL, Gillen DL, Williams JC, Jr., et al. A formal test of
the hypothesis that idiopathic calcium oxalate stones grow
PO
3BOEBMMhT
QMBRVF
#+6
*OU

46. Zarse CA, McAteer JA, Sommer AJ, et al. Nondestructive
analysis of urinary calculi using micro computed tomogra-
QIZ
#.$
6SPM

47. Abdel-Halim RE, Abdel-Halim MR. A review of urinary stone
BOBMZTJT
UFDIOJRVFT
4BVEJ
.FE
+

 Krambeck AE, Khan NF, Jackson ME, Lingeman JE, McA-
teer JA, Williams JC, Jr. Inaccurate reporting of mineral
composition by commercial stone analysis laboratories:
implications for infection and metabolic stones. J Urol.

49. Singh VK, Rai AK, Rai PK, Jindal PK. Cross-sectional study of
kidney stones by laser-induced breakdown spectroscopy.
-BTFST
.FE
4DJ

50. Lobinski R, Moulin C, Ortega R. Imaging and speciation
of trace elements in biological environment. Biochimie.

51. Austin C, Hare D, Rozelle AL, Robinson WH, Grimm R, Doble
P. Elemental bio-imaging of calcium phosphate crystal de-
posits in knee samples from arthritic patients. Metallomics.

52. Kontoyannis C, Bouropoulos N, Koutsoukos P. Urinary stone
layer analysis of mineral components by Raman spec-
troscopy, IR spectroscopy, and X-ray powder diffraction: a
DPNQBSBUJWF
TUVEZ
"QQMJFE
TQFDUSPTDPQZ

53. Cytron S, Kravchick S, Sela BA, et al. Fiberoptic infrared
spectroscopy: a novel tool for the analysis of urine and
VSJOBSZ
TBMUT
JO
TJUV
BOE
JO
SFBM
UJNF
6SPMPHZ

54. Fazil Marickar YM, Lekshmi PR, Varma L, Koshy P. EDAX
WFSTVT
'5*3
JO
NJYFE
TUPOFT
6SPM
3FT

55. 6MEBMM
"
"OBMZTJT
PG
VSJOBSZ
DBMDVMJ
B
RVBMJUZ
DPOUSPM
QSP-
HSBNNF
4DBOE
+
$MJO
-BC
*OWFTU

56. ,POKJLJ
5
JO
VSJOBSZ
DBMDVMJ
$BMDJG
5JTTVF
*OU

57. Materazzi S, Curini R, D'Ascenzo G, Magri A. TG-FTIR coupled
analysis applied to the studies in urolithiasis: characterization
PG
IVNBO
SFOBM
DBMDVMJ
5IFSNPDIJNJDB
BDUB