Research Article Understanding the Crystallinity Indices ...

Research ArticleUnderstanding the Crystallinity Indices Behavior ofBurned Bones and Teeth by ATR-IR and XRD in the Presence ofBioapatite Mixed with Other Phosphate and Carbonate Phases

Giampaolo Piga1 David Gonccedilalves234 T J U Thompson5 Antonio Brunetti1

Assumpcioacute Malgosa6 and Stefano Enzo7

1Department of Political Science Communication Engineering and Information Technologies University of SassariViale Mancini 5 07100 Sassari Italy2Research Centre for Anthropology and Health (CIAS) and Department of Life Sciences Universidade de CoimbraCalcada Martim Freitas 3000-456 Coimbra Portugal3Laboratory of Forensic Anthropology Department of Life Sciences Faculdade de Ciencias e Tecnologia Universidade de CoimbraCalcada Martim Freitas 3000-456 Coimbra Portugal4Laboratorio de Arqueociencias Direccao General do Patrimonio Cultural and LARCCIBIOInBIO Rua da Bica do Marques 21300-087 Lisbon Portugal5School of Science amp Engineering Teesside University Borough Road Middlesbrough TS1 3BA UK6Unitat drsquoAntropologia Biologica Department de Biologia Animal Biologia Vegetal i Ecologia Universitat Autonoma de BarcelonaEdifici C Bellaterra 08193 Barcelona Spain7Department of Chemistry and Pharmacy University of Sassari Via Vienna 2 07100 Sassari Italy

Correspondence should be addressed to Giampaolo Piga giapigaunissit

Received 4 November 2015 Accepted 3 January 2016

Academic Editor Jozef Kaiser

Copyright copy 2016 Giampaolo Piga et alThis is an open access article distributed under theCreative CommonsAttribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

We have critically investigated the ATR-IR spectroscopy data behavior of burned human teeth as opposed to the generally observedbehavior in humanbones thatwere subjected to heat treatment whether deliberate or accidental It is shown that the deterioration ofthe crystallinity index (CI) behavior sometimes observed in bones subjected to high temperature appears to be of higher frequencyin the case of bioapatite from teeth This occurs because the formation of the 120573-tricalcium phosphate (120573-TCP) phase otherwiseknown as whitlockite clearly ascertained by the X-ray diffraction (XRD) patterns collected on the same powdered specimensinvestigated by ATR-IRThese results point to the need of combining more than one physicochemical technique even if apparentlywell suitable in order to verify whether the assumed conditions assessed by spectroscopy are fully maintained in the specimensafter temperature andor mechanical processing

1 Introduction

Thestudy of burned human remains is of considerable impor-tance in archaeology forensic anthropology and crime sceneinvestigations We can have the presence of fire in many sit-uations such as accidents and homicides In fact fire is a com-mon method for attempting to conceal evidence of criminalactivity inflicted on human victims

To know the temperatures at which a bone was subjectedis a great index to better understand the modifications

suffered by bone structures due to combustion [1] to pro-mote the differentiation between natural and anthropogenicphenomena and to better interpret the techniques used inthe resolution of forensic cases where cremation or other firedamage to remains is present [2ndash6]

At this microscopic scale there are two key features influ-enced by heating that are worth exploring changes to the ele-mental composition and changes to the crystalline structureof the bone

Hindawi Publishing CorporationInternational Journal of SpectroscopyVolume 2016 Article ID 4810149 9 pageshttpdxdoiorg10115520164810149

2 International Journal of Spectroscopy

Thus new and accurate experimental methods areneeded to clarify the variety of factors that lead to varyinglevels of thermal effects

It has been argued that the most appropriate means ofaddressing microstructural studies of burned bones are thephysicochemical and spectroscopic approaches such as theFourier Transform Infrared Spectroscopy (FT-IR) and X-raydiffraction (XRD) [7ndash14] In recent years many researchershave turned their attention to alternative ways of studyingand identifying burned bones For this purpose the potentialof the crystallinity index (CI)mdashor splitting factor (SF)mdashhas been investigated intensively [9 15ndash22] although theapplications on real forensic or archaeological scenarios arestill rare in literature [20ndash23] CI can be measured with bothX-ray diffraction (XRD) and Fourier Transform InfraredSpectroscopy (FT-IR) Although both methods have beenused in the literature [20 21 23] the CI values defined usingXRD cannot be directly compared to those created using FT-IR [11 18 24]

XRD is ideal for defining a crystallinity parameter ofthe bioinorganic phase as the pattern involves directly theeffect of 3D periodicity (ie the organization degree in alldirections) of the elementary cell the smallest unit funda-mental for expressing the physical chemical and symmetryproperties of a crystal Conversely the FT-IR spectroscopysupplies a fingerprint of the chemical environment surround-ing the bond vibrations excited by the frequencies of the IRincoming beam Nevertheless it should be maintained thatFT-IR is advantageous to define a fresh bone because it isparticularly sensible to the presence of genetic matter as itcan be verified in the frequency range 1300ndash1700 cmminus1 [25]On the contrary hard X-ray radiation used in diffraction isrelatively insensitive to the presence of such component

FT-IR spectroscopy has the potential for being portableinto the field requires a very small amount of sample canbe cheaper to use and has been shown to be more accurateat lower burning temperatures [16 26 27] In particularthe methodology of KBr FT-IR for the sample preparationinvolves laborious dilution in a transparent means while withFT-IR ATR (attenuated total reflectance) the infrared beamimpinges directly a large area of the sample [18] maximizingthe reproducibility of the signals regardless the protocolsfollowed by the operator

However the accuracy of temperature determinationsolely using the FT-IR approach has some critical aspects Inthe absence of long range order information it is generallyassumed that the inorganic apatite component remains as asingle phase but this may not always be the case once thebone is subjected to a high-temperature treatment In factwhen bioapatite is subjected to a thermal treatment we canfind also amultiphase condition for the resultant product dueto a transformation of a part of hydroxylapatite (HA) to the120573-three-calcium-phosphate phase (120573-TCP) of a mineral namedwhitlockite [28]

The presence of 120573-TCP as well as the presence of othermineralogical phases due to various taphonomy effects canstrongly alter the shape of V

4

-V3

bands consequently thecalculation of the CI and all the recently proposed ratios

by Thompson et al [29] may not be correct That is why amultidisciplinary approach would be always advisable possi-bly with the combined use of various physicochemical tech-niques such as ATR-IR and XRD

In this sense we present in this paper a collection ofcomparative examples using animal and human bones com-bined with synthetic apatite heat-treated at selected temper-atures The trend of CI was inspected in order to identifyadvantages and limitations of the use of the FT-IR spectro-scopic technique in the study of burned bones

2 Material and Methods

Thehuman bones fragments and teeth employed in this studywere kindly made available from the Universitat Autonomade Barcelona (Spain) The cremated human teeth originatefrom the Necropolis of Monte Sirai (Carbonia Italy) Thepig bone specimens were kindly made available from theDepartment of Animal Biology University of Sassari (Italy)Synthetic powder hydroxylapatite was synthesized byAldrichChemistry

The samples were heat-treated in air using a NEY mufflefurnace at selected temperatures (500-700-900-1100∘C for 10and 40 minutes) using a rate of 20∘Cmin both for heatingand for cooling the bone specimens

FT-IR spectra were collected in ATRmode with a BrukerAlpha Platinum-ATR interferometer in terms of absorbanceversus wavenumber ] in the range 370ndash4000 cmminus1 with a res-olution of 4 cmminus1 Each spectrum was obtained by averaging256 interferograms The loose powder was dispersed inside ahole cavity of spheroidal shape with its surface aligned to theplate defining it

The crystallinity index adopted here is the same asthat used in the majority of archaeological and forensicapplicationsThe absorption bands at 605 and 565 cmminus1 wereused following baseline correction and the heights of theseabsorptions peaks were summed and then divided by theheight of the minimum between them [30]

The bone samples were analyzed with a Bruker M4Tornado 120583-XRF spectrometer using a Rh X-ray source modelMCBM50ndash06 Bworking at 50 kV and 600120583Aunder vacuum(20mbar) and using an Al filter 125 120583m thick In order tocheck the macroscopic chemical homogeneity a series of 20spectra were collected for each bone specimen Each spec-trum was accumulated for 600 s

The XRD patterns were collected using Bruker D2 Phaserinstrument working at a power of 30 kV and 10mA in theBragg-Brentano vertical alignment with a Cu-Ka tube emis-sion (120582 = 15418 A)

The width of divergent and antiscatter slits was 1mm(061∘) primary and secondary axial Soller slits of 25∘ werealso mounted with a linear detector LYNXEYE with 5∘opening and a monochromatisation by Ni foil for the Kbradiation The powder patterns were collected in the angularrange 9∘ndash140∘ in 2120579 with a step size of 005∘ The collectiontime of each pattern was pursued for 47min Our sampleholder for XRD analysis is a circular cavity of 25mm in

International Journal of Spectroscopy 3

400 800 1200 1600

ATR

tran

smitt

ance

(au

)

Not burned

Human bone

4(PO4)3minus

3(PO4)3minus

3(CO3)2minus

Frequency (cmminus1)

1100∘C

900∘C

700∘C500∘C

Figure 1 The ATR transmittance spectrum of a human unburnedbone (bottom pattern) and that for the same specimen treated at the500ndash1100∘C temperature range It is possible to appreciate a wholeconventional evolution of bands as discussed in the text

diameter and 2mm in depth containing about 190mg ofpowder bone

Digitized diagrams were initially subjected to preproc-essing for qualitative phase recognition according to theprograms Highscore and Match and then analyzed quan-titatively according to the Rietveld method [31] using theprogrammeMAUD [32] It is worth noting that one stringentrequirement of any Rietveld program needs the correct load-ing of the crystal structure solution of substances not onlyconcerning space group and lattice parameters but alsoincluding atomic location of the asymmetric unit [33]

3 Results and Discussion

In Figure 1 we report a conventional behavior of the FT-IR spectra collected in ATR mode as a function of reportedtemperature values (500 700 900 and 1100∘C for 10minutes)for a human bone

Summarizing briefly the V4

(PO4

) and V3

(PO4

) bandsoccur in the 500ndash700 and 850ndash1200 cmminus1 range respectivelyThe series of bands in the range 1300ndash1800 cmminus1 particularlythose at 1417 and 1660 cmminus1 are attributed to the presenceof carbonate groups in bone material and to the organicgenetic components respectively and can be used as a meansfor quantitative evaluation of their presence during drying[18 34]

Our spectra evolution in such range confirms the use-fulness and convenience of the proposed approach We canalso note increasing sharpening of the V

3

and V4

bandsattributed to phosphate groups In particular the V

4

bandsare further used as a means to study the CI evolution throughnumerical evaluation of the splitting factor SF in the pertinentrange selected as a function of temperature and reported inFigure 2

On this clear evidence the calibration of the CI asa function of temperature has been reversed in order to

500 600 700

ATR

tran

smitt

ance

(au

)

Not burned

Human bone


1100∘C

900∘C

700∘C

500∘C

SF = 592

SF = 482

SF = 451

SF = 300

SF = 296

Figure 2 The V4

(PO4

) bands are also magnified in the frequencyrange where the CI crystallinity index or SF splitting factor areworked out numerically and their figures reported

estimate the temperature to which a bone specimen wassubjected after a fire event [18ndash20]

It is obvious that the reliability of such results depends onthe validity of assumptions involved in the study underwayand strictly maintained by the sample under investigationIn particular the infrared spectroscopy is sensible to thevibrationsmodes around selected atomic species (or molecu-lar groups) in a solid crystalline matrix It is expected thatsuch matrix when subjected to thermal treatment enforcesits crystalline properties that is its degree of (3D) spatialorganization in the course of a thermal treatment This mayoccur in at least two different but concomitant ways (i)crystal growth and (ii) elimination of imperfections from theregular lattice In both cases it is implicitly assumed that themain parameters governing the crystal symmetry (ie thesymmetry operation ie the space group assumed by thecrystal) are not changing Whether this occurs or not can beinspected clearly by X-ray diffraction

Figure 3 shows the behavior of the ATR-IR spectra ofhuman teeth as a function of reported temperature values(500 700 900 and 1100∘C for 40 minutes) Tooth enamel isanother bioapatite product whose crystallinity study versustemperature could be addressed by ATR similar to humanbones Making reference to the range 1300ndash1800 cmminus1 wemay note better the carbonate bands (and their progressivedisappearance) rather than those of the genetic material hereless evident

Of course these bands are disappearing in the high-temperature spectra If we follow simultaneously the V

4

bandevolution we may envisage a complex progression of signals(see Figure 4)


500 1000 1500

ATR

tran

smitt

ance

(au

)Human teeth


1100∘C

900∘C

700∘C

500∘C

Figure 3 The spectra evolution as a function of temperaturetreatment for a human tooth shows an unconventional behavior forthe V4

(PO4

) band in terms of crystallinity index values We can alsonote an additional peak which appears at ca 1123 cmminus1 as indicatedby arrows

500 600 700

ATR

tran

smitt

ance

(au

)

Human teeth


1100∘C

900∘C

700∘C

500∘C

SF = 326

SF = 352

SF = 333

SF = 295

Figure 4 Magnification of the V4

band evolution we may envisagea complex progression of signals In this case the values of SF do notpresent an increasingmonotonic trendwith temperature but remainnearly constant around 33-34 Note the shoulder which appears atabout 547 cmminus1 attributable to 120573-TCP phase

In this case the values of SF do not present an increasingmonotonic trend with temperature but remain nearly con-stant around 33-34

As a matter of fact in the temperature range 700ndash1100∘Cfrom the XRD pattern we were able to recognize the presenceof 120573-TCP phase On account of the new system createdthe IR bands of 120573-TCP are expected to overlap with thebands of bioapatite An additional peak at ca 1123 cmminus1 (seeFigure 3) and the shoulder which appears at about 547 cmminus1

500 600 700 800

Tran

smitt

ance

(au

)

Not burned

Synthetic hydroxylapatite

Wavenumber (cmminus1)

1100∘C

900∘C

700∘C

500∘C

Figure 5 The ATR experiment involving synthetic hydroxylapatitedisplays the emergency of high-temperature features sensibly affect-ing the conventional band shape and sharpening to the point ofmaking any CI determination useless

(see Figure 4) as indicated by arrows are attributable to 120573-TCP

To this regard Figure 5 shows ATR-IR curves in cor-respondence with the V

4

(PO4

) band for the transformationbehavior of synthetic hydroxylapatite as a function of theindicated temperatures Although the curve of unburnedsynthetic hydroxylapatite shows features different fromuntreated bioapatite we may notice that from 700 up to1100∘C the high temperature bands with new componentsare heavily affected with respect to their original shape to thepoint of making any SF determination useless

Figures 6 7 and 8 refer to the changes of the V4

bandshape due to the presence of 120573-TCP at various levels in threedifferent specimens all of them subjected to high temperatureheat treatment supplemented with XRD patterns and corre-sponding Rietveld fit

Figure 6(a) shows a conventional pattern of a crematedhuman tooth from an archaeological cremationThe burningtemperature reached during the process is unknown Nev-ertheless the value of SF (568) permits estimating roughlythe temperature [18ndash20] Note that the peak at 562 cmminus1 isof intensity higher than that of the peak at 600 cmminus1 TheXRD phase analysis of Figure 6(b) shows that in addition tothe predominating presence of bioapatite (red curve) thereis a weak appearance (11 wt) of the 120573-TCP phase (bluecurve) Accordingly the bar sequences at the bottom markthe expected position of peaks for the indicated phases Thecurve below represents the residuals that is the differencebetween the square root of calculated and experimentalintensities respectively

Figure 7(a) shows a pig bone which was treated at1100∘C in a muffle furnace for 10 minutes With respect


500 600 700

ATR

tran

smitt

ance

(au

)Cremated human tooth562

599

632


SF = 568

(a)

X-ra

y in

tens

ity (a

u)

Cremated human tooth

89 bioapatite11 120573-TCP

28 30 32 34 36 3826Scattering angle 2120579

(b)

Figure 6 A detailed comparison of different FT-IR curves collected for three different bioapatite specimens with the correspondent XRDpatterns and relevant Rietveld phase evaluationThe distortions of ATR curves with respect to the conventional expected behavior are strictlyrelated to the amount of 120573-TCP phase which was stimulated by the high-temperature treatment as it clearly emerges from XRD analyses

ATR

tran

smitt

ance

(au

)

632

601568 SF = 361

550 600 650 700500Frequency (cmminus1)

Pig bone_1100∘C

(a)

X-ra

y in

tens

ity (a

u)


Pig bone_1100∘C


(b)


to the previous case we may notice a substantial changeof the band shape In addition the peaks at 568 cmminus1 and601 cmminus1 respectively show approximately the same heightAlso note that the prominent shoulder previously indicatedby the arrow in Figure 4 here is missing According to theRietveld fit of the XRD pattern (data points) this specimenhad developed 24 of 120573-TCP and 86 of bioapatite (seeFigure 7(b))

Figure 8(a) represents an extreme case in which a humantoothwas treated in a furnace at 1100∘C for 40minutes In thiscase the relative intensities of the peaks at 569 and 603 cmminus1respectively are reversed with respect to the curve recordedfor the specimen shown in Figure 6(a)

Analysis of the correspondent XRD pattern has estab-lished the presence of 70 120573-TCP phase for such spec-imen (see Figure 8(b)) Refined lattice parameters turned

out to be 1037 and 3723 A to compare with the values of119886 = 1042 and 3742 A respectively for pure commercial 120573-TCP (synthesized by Aldrich Chemistry)

The appearance of 120573-TCP from human bones seemsdifficult to account for [35] and has been related to the envi-ronmental pH andor to presence of magnesium ions whichmay substitute for calcium The transformation processinvolves multiple intermediates the stability of whichdepends on the cation (Ca andMg) activities and the solutionpH However in our studies of biomaterials such as bonesand teeth we never observed the 120573-TCP phase nor the clearpresence of magnesium ions from XRF spectroscopy

As is shown in Figure 9 the X-ray fluorescence spectradid not show evaluable peaks attributed to magnesium(125 keV) for a list of specimens where high-temperatureformation of 120573-TCP was reported As is also seen in Figure 6


ATR

tran

smitt

ance

(au

)

630

601569

SF = 319


Human tooth_1100∘C

(a)

X-ra

y in

tens

ity (a

u)




(b)


X-ra

y flu

ores

cenc

e (au

)

Mg

P

Ca

ClSAl

Pig bone

Crematedhuman tooth

2 3 4 5 61Energy (keV)

Human tooth1100∘C_10minHuman tooth1100∘C_40min

Figure 9 The XRF spectra analysis for bones and tooth wherewe have observed formation of 120573-TCP after a high temperaturetreatment The peak of magnesium expected at 125 keV appearsto be totally absent for all spectra This cast some doubts aboutthe presumed role of Mg ions in stabilizing the 120573-TCP phaseNevertheless there are other weak peaks from other elements (AlS and Cl) whose role should be inspected

there are other chemical elements such as chlorine sulphurandperhaps aluminium thatmay be relatedwith the observedtransformation A high-temperature treatment seems to bethe necessary requisite in order to observe conversion ofbioapatite to 120573-TCP [28]

In fact Elliott [36] has suggested for bioapatite a generalchemical equation of the type A

5

(BO4

)3

(X) where A = Ca2+or Mg2+ ions B = P or S and X = Fminus OHminus or (12)CO

3

2minus

When A = Ca2+ B = P(V) and X = OHminus we meet thechemical equation of synthetic apatite transforming to pure120573-TCP

2Ca5

(PO4

)3

OH 997888rarr 3Ca3

(PO4

)2

+ CaO +H2

O (1)

Of course in such high temperature reaction gaseous wateris supposed to be evolved which may account for sporadicobservations of teeth eruption and explosion during heat-treatment while solid CaO can rehydrate to Ca(OH)

2

(port-landite) after cooling down at room temperature

Unfortunately there is no well-defined temperature forthe above transformation reaction to occur the main reasonsbeing that apart frompH bioapatitemay be stabilized unpre-dictably also by grain boundaries defects and inclusion ofvarious chemical species difficult to identify and evaluateproperly

The appearance of 120573-TCP phase from bones appears tobe sporadic and seems to occur at temperatures around1100∘C [28] Conversely for teeth we have observed a moresystematic occurrence of 120573-TCP at temperatures as low as750∘C

Figure 10 shows the sequence of Rietveld fit for the humanteeth which were thermally treated in a furnace at 700 750900 and 1100∘C for 10 minutes at a rate of 20∘Cmin and thencooling in air The patterns show the appearance of the 120573-TCP phase (blue full line) occurring at a temperature as lowas 750∘C and following peak sharpening

A 120573-TCP amount of ca 5 from an otherwise bioapatitematrix can be ascertained from the XRD patterns in the2120579 range 26ndash37 using our experimental conditions payingattention to its most intense peak (0 2 10) occurring at 2120579= 3118∘ in reason of their lattice parameters 119886 = 1037 and 119888 =3721 A respectively

As can be seen in Figure 7 such shoulder emerges moreclearly with other diagnostic peaks of 120573-TCP at highertemperatures of treatment because of sharpening of thepeaks related to the increase of the average crystallite size for


X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C

32 3628Scattering angle 2120579

Figure 10 The series of XRD patterns for a tooth heat-treated at700ndash1100∘C temperature range suggest that the occurrence of120573-TCPis observable by heating the specimen in a furnace as low as 750∘Cfor 10 minutes at a rate of 20∘Cmin and then cooling in air

both bioapatite and 120573-TCP phases andor release of internalstrain

Two other points are worth noting fromRietveld analysis

(i) The amount of 120573-TCP separated at temperatures ashigh as 1100∘C remains approximately around 14

(ii) The lattice parameters of 120573-TCP remain essentiallyunchanged during the thermal process and the valuesare slightly below those determined in the literatureof the ldquosupposed magnesium stabilizedrdquo phase Com-parison with the XRD quantitative data reported inFigure 6(b) suggest that the holding time at a finaltemperature may help the kinetics of decomposi-tion at parity of temperature rate increase selectedFinally we would like to add that the unknown phasereported from thermal treatments of dental tissueand synthetic hydroxylapatite in the synchrotronradiation patterns of Sui et al [37] (120582 is not reportedbut calculable as ca 069 A) is very likely the 120573-TCPphase

We should note that such presence of120573-TCP is hardly distinctin the V

4

band of phosphates in the ATR-IR spectrum becauseof their relative importance in comparison to bioapatiteNevertheless the PndashO bond length distribution of phosphatein bioapatite [38] is certainly different from phosphate bondlength distribution in 120573-TCP [39]

4 Conclusions

Theuse of ATR-IR spectroscopy in the study of burned bonesusing the V

4

band of phosphates in the range 500ndash700 cmminus1may deserve special care in the case of occurrence of 120573-TCP

(or equivalently other phosphate phases that in principle canreact after chemical or physical treatments applied to thebioapatite) This is because the range of frequencies wherethe CI of bioapatite is determined turns out to be heavilyaffected by other bands of similar phosphate groups like thoseallocated in the 120573-TCP crystal structure This emphasize theuse of XRD as a valuable tool for supplementing the studiesby ATR-IR on human bones and teeth to assess or rejectoccurrence of high-temperature fires both in forensic and inarchaeological bone sample remains Even when the shapeof the V

4

band is different from the expected profile thecomparison between ATR-IR and XRD data may contributeto reconstruct the firing processes to which the bones weresubjected

For example the presence of 120573-TCP in human teeth asrevealed by XRD may be additional evidence of a thermaltreatment equivalent to at least 750∘C Normally the CIvalues from ATR-IR spectra conducted for estimating theheat treatment are correlated just to theXRDpeak sharpeningfrom the apatite phase throughout a line broadening analysisOn the other hand also analysis of the V

4

band of phosphatesdeviating from the conventional shape so far examined canbe interpreted more rigorously

The reason why 120573-TCP appears at relatively moder-ate temperature in teeth examined here in comparison tobones still remains obscure and further studies need to beaddressed acquiring information about chemical species andfollowing the crystal structure parameters

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors thank Professor Marco Zedda (Department ofAnimal Biology University of Sassari Italy) and Dr MicheleGuirguis (Department of History University of Sassari Italy)for supplying some osseous materials employed in this studyand Professor Plinio Innocenzi Dr Luca Malfatti and DrBarbara Lasio [Sciences and Nanotechnology Laboratory(LMNT) University of Sassari Italy] This work is supportedby Autonomous Region of Sardinia with the project titledldquoArchaeometric and Physico-Chemical Investigation Using aMulti-Technique Approach on Archaeological Anthropologicaland Paleontological Materials from the Mediterranean Areaand Sardiniardquo

References

[1] R J March ldquoLrsquoetude des structures de combustion prehisto-riques une approche interdisciplinairerdquo in XIII InternationalCongress of Prehistoric and protohistoric Sciences O Bar-JosefL Cavalli-Sforza R J March and Piperno Eds Colloquia 5The Lower and Middle Paleolithic Colloquium IX pp 251ndash275Forlı Italy September 1996


[2] D W Owsley ldquoIdentification of the fragmentary burnedremains of twoUS journalists seven years after their disappear-ance in Guatemalardquo Journal of Forensic Sciences vol 38 no 6pp 1372ndash1382 1993

[3] K A Murray and J C Rose ldquoThe analysis of cremains acase study involving the inappropriate disposal of mortuaryremainsrdquo Journal of Forensic Sciences vol 38 no 1 pp 98ndash1031993

[4] J L Holden J G Clement and P P Phakey ldquoAge and temper-ature related changes to the ultrastructure and composition ofhuman bone mineralrdquo Journal of Bone and Mineral Researchvol 10 no 9 pp 1400ndash1409 1995

[5] K A R Kennedy ldquoThewrong urn commingling of cremains inmortuary practicesrdquo Journal of Forensic Sciences vol 41 no 4pp 689ndash692 1996

[6] C Cattaneo S DiMartino S Scali O E Craig M Grandi andR J Sokol ldquoDetermining the human origin of fragments ofburnt bone a comparative study of histological immunologicaland DNA techniquesrdquo Forensic Science International vol 102no 2-3 pp 181ndash191 1999

[7] P Shipman G Foster and M Schoeninger ldquoBurnt bones andteeth an experimental study of color morphology crystalstructure and shrinkagerdquo Journal of Archaeological Science vol11 no 4 pp 307ndash325 1984

[8] H Newesely ldquoChemical stability of hydroxyapatite under dif-ferent conditionsrdquo in Trace Elements in Environmental HistoryProceedings of the Symposium held from June 24th to 26th 1987at Gottingen G Grupe and B Herrmann Eds Proceedings inLife Sciences pp 1ndash16 Springer Berlin Germany 1988

[9] M C Stiner S L Kuhn S Weiner and O Bar-YosefldquoDifferential burning recrystallization and fragmentation ofarchaeological bonerdquo Journal of Archaeological Science vol 22no 2 pp 223ndash237 1995

[10] A Ravaglioli A Krajewski G C Celotti et al ldquoMineralevolution of bonerdquo Biomaterials vol 17 no 6 pp 617ndash622 1996

[11] K D Rogers and P Daniels ldquoAn X-ray diffraction study ofthe effects of heat treatment on bone mineral microstructurerdquoBiomaterials vol 23 no 12 pp 2577ndash2585 2002

[12] G Piga A Malgosa T J U Thompson and S Enzo ldquoA newcalibration of the XRD technique for the study of archaeologicalburned human remainsrdquo Journal of Archaeological Science vol35 no 8 pp 2171ndash2178 2008

[13] G Piga T J U Thompson A Malgosa and S Enzo ldquoThepotential of X-ray diffraction in the analysis of burned remainsfrom forensic contextsrdquo Journal of Forensic Sciences vol 54 no3 pp 534ndash539 2009

[14] G Piga The use of spectroscopy and diffraction techniquesin the study of bones and implications in anthropology paleon-tology and forensic sciences [PhD thesis] Biblioteca deComunicacion y Hemeroteca General Universitat Autonomade Barcelona 2012 httpwwweducacionesteseomostrarRefdoref=996840

[15] H E C Koon R A Nicholson and M J Collins ldquoA practicalapproach to the identification of low temperature heated boneusing TEMrdquo Journal of Archaeological Science vol 30 no 11 pp1393ndash1399 2003

[16] L E Munro F J Longstaffe and C D White ldquoBurning andboiling ofmodern deer bone effects on crystallinity and oxygenisotope composition of bioapatite phosphaterdquo PalaeogeographyPalaeoclimatology Palaeoecology vol 249 no 1-2 pp 90ndash1022007

[17] J Olsen J Heinemeier P Bennike C Krause K MargretheHornstrup and H Thrane ldquoCharacterisation and blind testingof radiocarbon dating of cremated bonerdquo Journal of Archaeolog-ical Science vol 35 no 3 pp 791ndash800 2008

[18] T J UThompson M Gauthier and M Islam ldquoThe applicationof a newmethod of Fourier Transform Infrared Spectroscopy tothe analysis of burned bonerdquo Journal of Archaeological Sciencevol 36 no 3 pp 910ndash914 2009

[19] T J U Thompson M Islam K Piduru and A Marcel ldquoAninvestigation into the internal and external variables actingon crystallinity index using Fourier Transform Infrared Spec-troscopy on unaltered and burned bonerdquo PalaeogeographyPalaeoclimatology Palaeoecology vol 299 no 1-2 pp 168ndash1742011

[20] G Piga M Guirguis P Bartoloni A Malgosa and S EnzoldquoA funerary rite study of the Phoenician-Punic necropolis ofMount Sirai (Sardinia Italy)rdquo International Journal of Osteoar-chaeology vol 20 no 2 pp 144ndash157 2010

[21] G Piga A Malgosa T J U Thompson M Guirguis and SEnzo ldquoAunique case of prone position in the primary cremationTomb 252 ofMonte Sirai necropolis (Carbonia Sardinia Italy)rdquoInternational Journal of Osteoarchaeology vol 25 no 2 pp 146ndash159 2015

[22] K E Squires T J UThompsonM Islam and A ChamberlainldquoThe application of histomorphometry and Fourier TransformInfrared Spectroscopy to the analysis of early Anglo-Saxonburned bonerdquo Journal of Archaeological Science vol 38 no 9pp 2399ndash2409 2011

[23] G Piga J H Hernandez-Gasch AMalgosaM L Ganadu andS Enzo ldquoCremation practices coexisting at the SrsquoIllot des PorrosNecropolis during the Second Iron Age in the Balearic Islands(Spain)rdquo HOMOmdashJournal of Comparative Human Biology vol61 no 6 pp 440ndash452 2010

[24] S Chakraborty S Bag S Pal and A K Mukherjee ldquoStruc-tural and microstructural characterization of bioapatites andsynthetic hydroxyapatite using X-ray powder diffraction andFourier transform infrared techniquesrdquo Journal of AppliedCrystallography vol 39 no 3 pp 385ndash390 2006

[25] G Piga A Brunetti B Lasio S Enzo and A Malgosa ldquoXRFinvestigation on skeletal remains from King Peter III of Aragon(1239ndash1285AD) andQueenBlanche ofAnjou (1280ndash1310AD)rdquoApplied Physics A vol 114 no 3 pp 647ndash653 2014

[26] S Weiner P Goldberg and O Bar-Yosef ldquoBone preservationin Kebara Cave Israel using on-site Fourier transform infraredspectrometryrdquo Journal of Archaeological Science vol 20 no 6pp 613ndash627 1993

[27] L E Wright and H P Schwarcz ldquoInfrared and isotopicevidence for diagenesis of bone apatite at Dos Pilas Guatemalapalaeodietary implicationsrdquo Journal of Archaeological Sciencevol 23 no 6 pp 939ndash944 1996

[28] G Piga G Solinas T J U Thompson A Brunetti A Malgosaand S Enzo ldquoIs X-ray diffraction able to distinguish betweenanimal and human bonesrdquo Journal of Archaeological Sciencevol 40 no 1 pp 778ndash785 2013

[29] T J UThompsonM Islam andM Bonniere ldquoA new statisticalapproach for determining the crystallinity of heat-altered bonemineral from FTIR spectrardquo Journal of Archaeological Sciencevol 40 no 1 pp 416ndash422 2013

[30] S Weiner and O Bar-Yosef ldquoStates of preservation of bonesfrom prehistoric sites in the Near East a surveyrdquo Journal ofArchaeological Science vol 17 no 2 pp 187ndash196 1990


[31] H M Rietveld ldquoLine profiles of neutron powder-diffractionpeaks for structure refinementrdquo Acta Crystallographica vol 22no 1 pp 151ndash152 1967

[32] L Lutterotti ldquoTotal pattern fitting for the combined size-strain-stress-texture determination in thin film diffractionrdquo NuclearInstruments and Methods in Physics Research Section B BeamInteractions withMaterials andAtoms vol 268 no 3-4 pp 334ndash340 2010

[33] S Graulis D Chateigner R T Downs et al ldquoCrystallographyOpen Databasemdashan open-access collection of crystal struc-turesrdquo Journal of Applied Crystallography vol 42 no 4 pp 726ndash729 2009

[34] M Lebon I Reiche F Frohlich J-J Bahain and C FalgueresldquoCharacterization of archaeological burnt bones contributionof a new analytical protocol based on derivative FTIR spec-troscopy and curve fitting of the V

1

V3

PO4

domainrdquo Analyticaland Bioanalytical Chemistry vol 392 no 7 pp 1479ndash1488 2008

[35] H L JangHK Lee K JinH-YAhnH-E Lee andKTNamldquoPhase transformation from hydroxyapatite to the secondarybone mineral whitlockiterdquo Journal of Materials Chemistry Bvol 3 no 7 pp 1342ndash1349 2015

[36] J C Elliott Structure and Chemistry of the Apatites and OtherCalciumOrthophosphates Elsevier ScienceampTechnology Ams-terdam The Netherlands 1994

[37] T Sui M A Sandholzer A J G Lunt et al ldquoIn situ X-rayscattering evaluation of heatinduced ultrastructural changes indental tissues and synthetic hydroxyapatiterdquo Journal of the RoyalSociety Interface vol 11 no 95 Article ID 20130928 2014

[38] R M Wilson J C Elliott and S E P Dowker ldquoRietveldrefinement of the crystallographic structure of human dentalenamel apatitesrdquoAmericanMineralogist vol 84 no 9 pp 1406ndash1414 1999

[39] MYashimaA Sakai T Kamiyama andAHoshikawa ldquoCrystalstructure analysis of 120573-tricalcium phosphate Ca

3

(PO4

)2

byneutron powder diffractionrdquo Journal of Solid State Chemistryvol 175 no 2 pp 272ndash277 2003

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


Thus new and accurate experimental methods areneeded to clarify the variety of factors that lead to varyinglevels of thermal effects

It has been argued that the most appropriate means ofaddressing microstructural studies of burned bones are thephysicochemical and spectroscopic approaches such as theFourier Transform Infrared Spectroscopy (FT-IR) and X-raydiffraction (XRD) [7ndash14] In recent years many researchershave turned their attention to alternative ways of studyingand identifying burned bones For this purpose the potentialof the crystallinity index (CI)mdashor splitting factor (SF)mdashhas been investigated intensively [9 15ndash22] although theapplications on real forensic or archaeological scenarios arestill rare in literature [20ndash23] CI can be measured with bothX-ray diffraction (XRD) and Fourier Transform InfraredSpectroscopy (FT-IR) Although both methods have beenused in the literature [20 21 23] the CI values defined usingXRD cannot be directly compared to those created using FT-IR [11 18 24]

XRD is ideal for defining a crystallinity parameter ofthe bioinorganic phase as the pattern involves directly theeffect of 3D periodicity (ie the organization degree in alldirections) of the elementary cell the smallest unit funda-mental for expressing the physical chemical and symmetryproperties of a crystal Conversely the FT-IR spectroscopysupplies a fingerprint of the chemical environment surround-ing the bond vibrations excited by the frequencies of the IRincoming beam Nevertheless it should be maintained thatFT-IR is advantageous to define a fresh bone because it isparticularly sensible to the presence of genetic matter as itcan be verified in the frequency range 1300ndash1700 cmminus1 [25]On the contrary hard X-ray radiation used in diffraction isrelatively insensitive to the presence of such component

FT-IR spectroscopy has the potential for being portableinto the field requires a very small amount of sample canbe cheaper to use and has been shown to be more accurateat lower burning temperatures [16 26 27] In particularthe methodology of KBr FT-IR for the sample preparationinvolves laborious dilution in a transparent means while withFT-IR ATR (attenuated total reflectance) the infrared beamimpinges directly a large area of the sample [18] maximizingthe reproducibility of the signals regardless the protocolsfollowed by the operator

However the accuracy of temperature determinationsolely using the FT-IR approach has some critical aspects Inthe absence of long range order information it is generallyassumed that the inorganic apatite component remains as asingle phase but this may not always be the case once thebone is subjected to a high-temperature treatment In factwhen bioapatite is subjected to a thermal treatment we canfind also amultiphase condition for the resultant product dueto a transformation of a part of hydroxylapatite (HA) to the120573-three-calcium-phosphate phase (120573-TCP) of a mineral namedwhitlockite [28]

The presence of 120573-TCP as well as the presence of othermineralogical phases due to various taphonomy effects canstrongly alter the shape of V

4

-V3

bands consequently thecalculation of the CI and all the recently proposed ratios

by Thompson et al [29] may not be correct That is why amultidisciplinary approach would be always advisable possi-bly with the combined use of various physicochemical tech-niques such as ATR-IR and XRD

In this sense we present in this paper a collection ofcomparative examples using animal and human bones com-bined with synthetic apatite heat-treated at selected temper-atures The trend of CI was inspected in order to identifyadvantages and limitations of the use of the FT-IR spectro-scopic technique in the study of burned bones

2 Material and Methods

Thehuman bones fragments and teeth employed in this studywere kindly made available from the Universitat Autonomade Barcelona (Spain) The cremated human teeth originatefrom the Necropolis of Monte Sirai (Carbonia Italy) Thepig bone specimens were kindly made available from theDepartment of Animal Biology University of Sassari (Italy)Synthetic powder hydroxylapatite was synthesized byAldrichChemistry

The samples were heat-treated in air using a NEY mufflefurnace at selected temperatures (500-700-900-1100∘C for 10and 40 minutes) using a rate of 20∘Cmin both for heatingand for cooling the bone specimens

FT-IR spectra were collected in ATRmode with a BrukerAlpha Platinum-ATR interferometer in terms of absorbanceversus wavenumber ] in the range 370ndash4000 cmminus1 with a res-olution of 4 cmminus1 Each spectrum was obtained by averaging256 interferograms The loose powder was dispersed inside ahole cavity of spheroidal shape with its surface aligned to theplate defining it

The crystallinity index adopted here is the same asthat used in the majority of archaeological and forensicapplicationsThe absorption bands at 605 and 565 cmminus1 wereused following baseline correction and the heights of theseabsorptions peaks were summed and then divided by theheight of the minimum between them [30]

The bone samples were analyzed with a Bruker M4Tornado 120583-XRF spectrometer using a Rh X-ray source modelMCBM50ndash06 Bworking at 50 kV and 600120583Aunder vacuum(20mbar) and using an Al filter 125 120583m thick In order tocheck the macroscopic chemical homogeneity a series of 20spectra were collected for each bone specimen Each spec-trum was accumulated for 600 s

The XRD patterns were collected using Bruker D2 Phaserinstrument working at a power of 30 kV and 10mA in theBragg-Brentano vertical alignment with a Cu-Ka tube emis-sion (120582 = 15418 A)

The width of divergent and antiscatter slits was 1mm(061∘) primary and secondary axial Soller slits of 25∘ werealso mounted with a linear detector LYNXEYE with 5∘opening and a monochromatisation by Ni foil for the Kbradiation The powder patterns were collected in the angularrange 9∘ndash140∘ in 2120579 with a step size of 005∘ The collectiontime of each pattern was pursued for 47min Our sampleholder for XRD analysis is a circular cavity of 25mm in


400 800 1200 1600

ATR

tran

smitt

ance

(au

)

Not burned

Human bone

4(PO4)3minus

3(PO4)3minus

3(CO3)2minus


1100∘C

900∘C

700∘C500∘C







(PO4

) and V3

(PO4



3

and V4


4



500 600 700

ATR

tran

smitt

ance

(au

)

Not burned

Human bone


1100∘C

900∘C

700∘C

500∘C

SF = 592

SF = 482

SF = 451

SF = 300

SF = 296

Figure 2 The V4

(PO4






4



500 1000 1500

ATR

tran

smitt

ance

(au

)Human teeth


1100∘C

900∘C

700∘C

500∘C


(PO4


500 600 700

ATR

tran

smitt

ance

(au

)

Human teeth


1100∘C

900∘C

700∘C

500∘C

SF = 326

SF = 352

SF = 333

SF = 295





500 600 700 800

Tran

smitt

ance

(au

)

Not burned



1100∘C

900∘C

700∘C

500∘C




4

(PO4







500 600 700

ATR

tran

smitt

ance

(au


599

632


SF = 568

(a)

X-ra

y in

tens

ity (a

u)




(b)


ATR

tran

smitt

ance

(au

)

632

601568 SF = 361


Pig bone_1100∘C

(a)

X-ra

y in

tens

ity (a

u)


Pig bone_1100∘C


(b)









ATR

tran

smitt

ance

(au

)

630

601569

SF = 319



(a)

X-ra

y in

tens

ity (a

u)




(b)


X-ra

y flu

ores

cenc

e (au

)

Mg

P

Ca

ClSAl

Pig bone

Crematedhuman tooth






5

(BO4

)3


3

2minus


2Ca5

(PO4

)3

OH 997888rarr 3Ca3

(PO4

)2

+ CaO +H2

O (1)


2








X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C








4


4 Conclusions


4



4



4





Acknowledgments


References





































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


400 800 1200 1600

ATR

tran

smitt

ance

(au

)

Not burned

Human bone

4(PO4)3minus

3(PO4)3minus

3(CO3)2minus


1100∘C

900∘C

700∘C500∘C







(PO4

) and V3

(PO4



3

and V4


4



500 600 700

ATR

tran

smitt

ance

(au

)

Not burned

Human bone


1100∘C

900∘C

700∘C

500∘C

SF = 592

SF = 482

SF = 451

SF = 300

SF = 296

Figure 2 The V4

(PO4






4



500 1000 1500

ATR

tran

smitt

ance

(au

)Human teeth


1100∘C

900∘C

700∘C

500∘C


(PO4


500 600 700

ATR

tran

smitt

ance

(au

)

Human teeth


1100∘C

900∘C

700∘C

500∘C

SF = 326

SF = 352

SF = 333

SF = 295





500 600 700 800

Tran

smitt

ance

(au

)

Not burned



1100∘C

900∘C

700∘C

500∘C




4

(PO4







500 600 700

ATR

tran

smitt

ance

(au


599

632


SF = 568

(a)

X-ra

y in

tens

ity (a

u)




(b)


ATR

tran

smitt

ance

(au

)

632

601568 SF = 361


Pig bone_1100∘C

(a)

X-ra

y in

tens

ity (a

u)


Pig bone_1100∘C


(b)









ATR

tran

smitt

ance

(au

)

630

601569

SF = 319



(a)

X-ra

y in

tens

ity (a

u)




(b)


X-ra

y flu

ores

cenc

e (au

)

Mg

P

Ca

ClSAl

Pig bone

Crematedhuman tooth






5

(BO4

)3


3

2minus


2Ca5

(PO4

)3

OH 997888rarr 3Ca3

(PO4

)2

+ CaO +H2

O (1)


2








X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C








4


4 Conclusions


4



4



4





Acknowledgments


References





































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


500 1000 1500

ATR

tran

smitt

ance

(au

)Human teeth


1100∘C

900∘C

700∘C

500∘C


(PO4


500 600 700

ATR

tran

smitt

ance

(au

)

Human teeth


1100∘C

900∘C

700∘C

500∘C

SF = 326

SF = 352

SF = 333

SF = 295





500 600 700 800

Tran

smitt

ance

(au

)

Not burned



1100∘C

900∘C

700∘C

500∘C




4

(PO4







500 600 700

ATR

tran

smitt

ance

(au


599

632


SF = 568

(a)

X-ra

y in

tens

ity (a

u)




(b)


ATR

tran

smitt

ance

(au

)

632

601568 SF = 361


Pig bone_1100∘C

(a)

X-ra

y in

tens

ity (a

u)


Pig bone_1100∘C


(b)









ATR

tran

smitt

ance

(au

)

630

601569

SF = 319



(a)

X-ra

y in

tens

ity (a

u)




(b)


X-ra

y flu

ores

cenc

e (au

)

Mg

P

Ca

ClSAl

Pig bone

Crematedhuman tooth






5

(BO4

)3


3

2minus


2Ca5

(PO4

)3

OH 997888rarr 3Ca3

(PO4

)2

+ CaO +H2

O (1)


2








X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C








4


4 Conclusions


4



4



4





Acknowledgments


References





































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


500 600 700

ATR

tran

smitt

ance

(au


599

632


SF = 568

(a)

X-ra

y in

tens

ity (a

u)




(b)


ATR

tran

smitt

ance

(au

)

632

601568 SF = 361


Pig bone_1100∘C

(a)

X-ra

y in

tens

ity (a

u)


Pig bone_1100∘C


(b)









ATR

tran

smitt

ance

(au

)

630

601569

SF = 319



(a)

X-ra

y in

tens

ity (a

u)




(b)


X-ra

y flu

ores

cenc

e (au

)

Mg

P

Ca

ClSAl

Pig bone

Crematedhuman tooth






5

(BO4

)3


3

2minus


2Ca5

(PO4

)3

OH 997888rarr 3Ca3

(PO4

)2

+ CaO +H2

O (1)


2








X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C








4


4 Conclusions


4



4



4





Acknowledgments


References





































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


ATR

tran

smitt

ance

(au

)

630

601569

SF = 319



(a)

X-ra

y in

tens

ity (a

u)




(b)


X-ra

y flu

ores

cenc

e (au

)

Mg

P

Ca

ClSAl

Pig bone

Crematedhuman tooth






5

(BO4

)3


3

2minus


2Ca5

(PO4

)3

OH 997888rarr 3Ca3

(PO4

)2

+ CaO +H2

O (1)


2








X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C








4


4 Conclusions


4



4



4





Acknowledgments


References





































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


X-ra

y in

tens

ity (a

u)

1100∘C

900∘C

750∘C

700∘C








4


4 Conclusions


4



4



4





Acknowledgments


References





































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of




































1

V3

PO4







3

(PO4

)2











Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of










Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

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