Trends in High Performance Liquid Chromatography for Cultural Heritage

  • Ilaria DeganoEmail author
  • Jacopo La Nasa
Part of the following topical collections:
  1. Analytical Chemistry for Cultural Heritage


The separation, detection and quantitation of specific species contained in a sample in the field of Cultural Heritage requires selective, sensitive and reliable methods. Procedures based on liquid chromatography fulfil these requirements and offer a wide range of applicability in terms of analyte types and concentration range. The main applications of High Performance Liquid Chromatography in this field are related to the separation and detection of dyestuffs in archaeological materials and paint samples by reversed-phase liquid chromatography with suitable detectors. The relevant literature will be revised, with particular attention to sample treatment strategies and future developments. Reversed phase chromatography has also recently gained increasing importance in the analysis of lipid binders and lipid materials in archaeological residues: the main advantages and disadvantages of the new approaches will be discussed. Finally, the main applications of ion chromatography and size exclusion chromatography in the field of Cultural Heritage will be revised in this chapter.


HPLC Ion chromatography Size exclusion chromatography RP-HPLC Black crusts Polymers in conservation Dyes Lipid binders 



The authors graciously acknowledge the National project “PRIN 2010–2011: Sustainability in cultural heritage: from diagnosis to the development of innovative systems for consolidation, cleaning and protection”, granted by MIUR for financial support. They would also like to acknowledge their colleagues at University of Pisa, professor Erika Ribechini and professor Francesca Modugno, for the fruitful discussions.


  1. 1.
    The chromatography and sample preparation terminology guide (2013)Google Scholar
  2. 2.
    Colombini MP, Modugno F (2009) Organic mass spectrometry in art and archeology. Wiley, ChichesterGoogle Scholar
  3. 3.
    Lomax SQ, Learner T (2006) A review of the classes, structures, and methods of analysis of synthetic organic pigments. J Am Inst Conserv 45(2):107–125CrossRefGoogle Scholar
  4. 4.
    Nowik W, Desrosiers S, Surowiec I, Trojanowicz M (2005) The analysis of dyestuffs from first- to second-century textile artefacts found in the Martresde-Veyre (France) excavations. Archaeometry 47:835–848CrossRefGoogle Scholar
  5. 5.
    Surowiec I (2008) Application of high-performance separation techniques in archaeometry. Microchim Acta 162:289–302CrossRefGoogle Scholar
  6. 6.
    Rosenberg E (2008) Characterisation of historical organic dyestuffs by liquid chromatography–mass spectrometry. Anal Bioanal Chem 391:33–57CrossRefGoogle Scholar
  7. 7.
    Degano I, Ribechini E, Modugno F, Colombini MP (2009) Analytical methods for the characterization of organic dyes in artworks and in historical textiles. Appl Spectrosc Rev 44:363–410CrossRefGoogle Scholar
  8. 8.
    Pauk V, Bartak P, Lemr K (2014) Characterization of natural organic colorants in historical and art objects by high-performance liquid chromatography. J Sep Sci 37:3393–3410CrossRefGoogle Scholar
  9. 9.
    Russell J, Singer BW, Perry JJ, Bacon A (2011) The identification of synthetic organic pigments in modern paints and modern paintings using pyrolysis-gas chromatography–mass spectrometry. Anal Bioanal Chem 400:1473–1491CrossRefGoogle Scholar
  10. 10.
    Ghelardi E, Degano I, Colombini MP, Mazurek J, Schilling M, Learner T (2015) Py-GC/MS applied to the analysis of synthetic organic pigments: characterization and identification in paint samples. Anal Bioanal Chem 407:1415–1431CrossRefGoogle Scholar
  11. 11.
    Carlesi S et al (2015) Discovering “The Italian Flag” by Fernando Melani (1907–1985). Spectrochim Acta Part A Mol Biomol SpectroscGoogle Scholar
  12. 12.
    Lech K, Wilicka E, Witowska-Jarosz J, Jarosz M (2013) Early synthetic dyes—a challenge for tandem mass spectrometry. J Mass Spectrom 48:141–147CrossRefGoogle Scholar
  13. 13.
    van Bommel MR, Vanden Berghe I, Wallert AM (2007) High-performance liquid chromatography and non-destructive three-dimensional fluorescence analysis of early synthetic dyes. J Chromatogr A 1157:260–272CrossRefGoogle Scholar
  14. 14.
    Confortin D, Neevel H, Brustolon M, Franco L, Kettelarij AJ, Williams RM, van Bommel MR (2010) Crystal violet: study of the photo-fading of an early synthetic dye in aqueous solution and on paper with HPLC-PDA, LCMS and FORS. J Phys Conf Ser 231:1–9CrossRefGoogle Scholar
  15. 15.
    de Keijzer M, van Bommell MR, Hofmann-de Keijzer R, Knaller R, Oberhumer E (2012). Indigo carmine: understanding a problematic blue dye. In: Contributions to the Vienna Congress 2012 (Vienna 2012), pp S87–S95Google Scholar
  16. 16.
    Restivo A, Degano I, Ribechini E, Colombini MP (2014) Development and optimisation of an HPLC-DAD-ESI-Q-TOF method for the determination of phenolic acids and derivatives. Plos One 9:e88762CrossRefGoogle Scholar
  17. 17.
    Halpine SM (1995) An improved dye and lake pigment analysis method for high performance liquid chromatography and diode-array detection. Stud Conserv 41:76–94Google Scholar
  18. 18.
    Nowik W, Marcinowska R, Kusyk K, Cardonc D, Trojanowicz M (2011) High performance liquid chromatography of slightly soluble brominated indigoids from Tyrian purple. J Chromatogr A 1218:1244–1252CrossRefGoogle Scholar
  19. 19.
    Surowiec I, Quye A, Trojanowicz M (2006) Liquid chromatography determination of natural dyes in extracts from historical Scottish textiles excavated from peat bogs. J Chromatogr A 1112:209–217CrossRefGoogle Scholar
  20. 20.
    Taujenis L, Olšauskaitė V (2012) Identification of main constituents of historical textile dyes by ultra performance liquid chromatography with photodiode array detection. Chemija 23:210–215Google Scholar
  21. 21.
    Serrano A, van Bommel M, Hallett J (2013) Evaluation between ultrahigh pressure liquid chromatography andhigh-performance liquid chromatography analytical methods for characterizing natural dyestuffs. J Chromatogr A 1318:102–111CrossRefGoogle Scholar
  22. 22.
    Troalen LG, Phillips AS, Peggie DA, Barran PE, Hulme AN (2014) Historical textile dyeing with Genista tinctoria L.: a comprehensive study by UPLC-MS/MS analysis. Anal Methods 6:8915–8923CrossRefGoogle Scholar
  23. 23.
    Wouters J, Grzywacz CM, Claro A (2011) A comparative investigation of hydrolysis methods to analyze natural organic dyes by HPLC-PDA. Stud Conserv 56:231–249CrossRefGoogle Scholar
  24. 24.
    Manhita A, Ferreira T, Candeias A, Barrocas Dias C (2011) Extracting natural dyes from wool—an evaluation of extraction methods. Anal Bioanal Chem 400:1501–1514CrossRefGoogle Scholar
  25. 25.
    Valianou L, Karapanagiotis I, Chryssoulakis Y (2009) Comparison of extraction methods for the analysis of natural dyes in historical textiles by high-performance liquid chromatography. Anal Bioanal Chem 395:2175–2189CrossRefGoogle Scholar
  26. 26.
    Sanyova J (2008) Mild extraction of dyes by hydrofluoric acid in routine analysis of historical paint micro-samples. Microchim Acta 162:361–370CrossRefGoogle Scholar
  27. 27.
    Blanc R, Espejo T, Lopez-Montes A, Torres D, Crovetto G, Navalon A, Vilchez JL (2006) Sampling and identification of natural dyes in historical maps and drawings by liquid chromatography with diode-array detection. J Chromatogr A 1122:105–113CrossRefGoogle Scholar
  28. 28.
    Sanyova J, Reisse J (2006) Development of a mild method for the extraction of anthraquinones from their aluminum complexes in madder lakes prior to HPLC analysis. J Cult Herit 7:229–235CrossRefGoogle Scholar
  29. 29.
    Lech K, Jarosz M (2011) Novel methodology for the extraction and identification of natural dyestuffs in historical textiles by HPLC–UV–Vis–ESI–MS. Case study: chasubles from the Wawel Cathedral collection. Anal Bioanal Chem 399:3241–3251CrossRefGoogle Scholar
  30. 30.
    Mouri C, Laursen R (2012) Identification of anthraquinone markers for distinguishing Rubia species in madder-dyed textiles by HPLC. Microchim Acta 179:105–113CrossRefGoogle Scholar
  31. 31.
    Zhang X, Laursen RA (2005) Development of mild extraction methods for the analysis of natural dyes in textiles of historical interest using LC-diode array detector-MS. Anal Chem 77:2022–2025CrossRefGoogle Scholar
  32. 32.
    VandenBerghe I, Gleba M, Mannering U (2009) Towards the identification of dyestuffs in early iron age Scandinavian peat bog textiles. J Archaeol Sci 36:1910–1921CrossRefGoogle Scholar
  33. 33.
    Colombini MP, Carmignani A, Modugno F et al (2004) Integrated analytical techniques for the study of ancient Greek polychromy. Talanta 63:839–848CrossRefGoogle Scholar
  34. 34.
    Stolyhwo A, Colin H, Guiochon G (1985) Analysis of triglycerides in oils and fats by liquid chromatography with the laser light scattering detector. Anal Chem 57:1342–1354CrossRefGoogle Scholar
  35. 35.
    Herrera LC, Potvin M, Melanson J (2010) Quantitative analysis of positional isomers of triacylglycerols via electrospray ionization tandem mass spectrometry of sodiated adducts. Rapid Commun Mass Spectrom 24:2745–2752CrossRefGoogle Scholar
  36. 36.
    Byrdwell WC, Neff WE (2002) Dual parallel electrospray ionization and atmospheric pressure chemical ionization mass spectrometry (MS), MS/MS and MS/MS/MS for the analysis of triacylglycerols and triacylglycerol oxidation products. Rapid Commun Mass Spectrom 16:300–319CrossRefGoogle Scholar
  37. 37.
    La Nasa J, Ghelardi E, Degano I, Modugno F, Colombini MP (2013) Core shell stationary phases for a novel separation of triglycerides in plant oils by high performance liquid chromatography with electrospray-quadrupole-time of flight mass spectrometer. J Chromatogr A 1308:114–124CrossRefGoogle Scholar
  38. 38.
    Saliu F, Degano I, Colombini MP (2014) Identification of triacylglycerols in archaelogical organic residues by core–shell reversed phase liquid chromatography coupled to electrospray ionization-quadrupole-time of flight mass spectrometry. J Chromatogr A 1346:78–87CrossRefGoogle Scholar
  39. 39.
    Mottram HR, Evershed RP (1996) Structure analysis of triacylglycerol positional isomers using atmospheric pressure chemical ionisation mass spectrometry. Tetrahedron Lett 37:8593–8596CrossRefGoogle Scholar
  40. 40.
    Kimpe K, Jacobs PA, Waelkens M (2002) Mass spectrometric methods prove the use of beeswax and ruminant fat in late Roman cooking pots. J Chromatogr A 968:151–160CrossRefGoogle Scholar
  41. 41.
    Kimpe K, Jacobs PA, Waelkens M (2001) Analysis of oil used in late Roman oil lamps with different mass spectrometric techniques revealed the presence of predominantly olive oil together with traces of animal fat. J Chromatogr A 937:87–95CrossRefGoogle Scholar
  42. 42.
    Romanus K, Van Neer W, Marinova E et al (2008) Brassicaceae seed oil identified as illuminant in Nilotic shells from a first millennium AD Coptic church in Bawit, Egypt. Anal Bioanal Chem 390:783–793CrossRefGoogle Scholar
  43. 43.
    Romanus K, Poblome J, Verbeke K, Luypaerts A, Jacobs P, De Vos D, Walekens M (2007) An evaluation of analytical and interpretative methodologies for the extraction and identification of lipids associated with pottery sherds from the site of Sagalassos, Turkey. Archaeometry 49:729–747CrossRefGoogle Scholar
  44. 44.
    Saliu F, Modugno F, Orlandi M, Colombini MP (2011) HPLC APCI–MS analysis of triacylglycerols (TAGs) in historical pharmaceutical ointments from the eighteenth century. Anal Bioanal Chem 401:1785–1800CrossRefGoogle Scholar
  45. 45.
    Berg JDJvd, Vermist ND, Carlyle L, Holcapek M, Boon JJ (2004) Effetcs of traditional processing methods of linseed oil on the composition of its triacylglcyerols. J Sep Sci 27:181–199Google Scholar
  46. 46.
    Chester TL (2013) Recent developments in high-performance liquid chromatography stationary phases. Anal Chem 85:579–589CrossRefGoogle Scholar
  47. 47.
    LaNasa J, Zanaboni M, Uldanck D, Degano I, Modugno F, Kutzke H, Tveit ES, Topalova-Casadiego B, Colombini mp (2015) Novel application of liquid chromatography/mass spectrometry for the characterization of drying oils in art: Elucidation on the composition of original paint materials used by Edvard Munch (1863–1944). Anal Chim Acta 896:177–189CrossRefGoogle Scholar
  48. 48.
    Nasa JL, Degano I, Modugno F, Colombini MP (2013) Alkyd paints in art: characterization using integrated massspectrometry. Anal Chim Acta 797:4–80Google Scholar
  49. 49.
    Nasa JL, Degano I, Modugno F, Colombini MP (2015) Industrial alkyd resins: characterization of pentaerythritol and phthalic acid esters using integrated mass spectrometry. Rapid Commun Mass Spectrom 29:225–237CrossRefGoogle Scholar
  50. 50.
    Nasa JL, Degano I, Modugno F, Colombini MP (2014) Effects of acetic acid vapour on the ageing of alkyd paint layers: multianalytical approach for the evaluation of the degradation processes. Polym Degrad Stab 105:257–264CrossRefGoogle Scholar
  51. 51.
    Gobbi G, Zappia G, Sabbioni C (1995) Anion determination in damage layers of stone monuments. Atmos Environ 29:703–707CrossRefGoogle Scholar
  52. 52.
    Sabbioni C, Ghedini N, Bonazza A (2003) Organic anions in damage layers on monuments and buildings. Atmos Environ 37:1261–1269CrossRefGoogle Scholar
  53. 53.
    Bonazza A, Sabbioni C, Ghedini N (2005) Quantitative data on carbon fractions in interpretation of black crusts and soiling on European built heritage. Atmos Environ 39:2607–2618CrossRefGoogle Scholar
  54. 54.
    Moropoulou A, Koui M, Kourteli Ch, Theoulakis P, Avdelidis NP (2001) Integrated methodology for measuring and monitoring salt decay in the medieval city of Rhodes porous stone. Mediterr Archaeol Archaeom 1:57–68Google Scholar
  55. 55.
    Tittarelli F, Moriconi G, Bonazza A (2008) Atmospheric deterioration of cement plaster in a building exposed to a urban environment. J Cult Herit 9:203–206CrossRefGoogle Scholar
  56. 56.
    Nava S, Becherini F, Bernardi A et al (2010) An integrated approach to assess air pollution threats to cultural heritage in a semi-confined environment: the case study of Michelozzo’s Courtyard in Florence (Italy). Sci Total Environ 408:1403–1413CrossRefGoogle Scholar
  57. 57.
    Fassina V, Favaro M, Naccari A, Pigo M (2002) Evaluation of compatibility and durability of a hydraulic lime-based plaster applied on brick wall masonry of historical buildings affected by rising damp phenomena. J Cult Herit 3:45–51CrossRefGoogle Scholar
  58. 58.
    Smolik J, Maskova L, Zikova N, Ondrackova L, Ondracek (2013) Deposition of suspended fine particulate matter in a library. Herit Sci 1:7CrossRefGoogle Scholar
  59. 59.
    Ghedini N, Ozga I, Bonazza A, Dilillo M, Cachier H, Sabbioni C (2011) Atmospheric aerosol monitoring as a strategy for the preventive conservation of urban monumental heritage: the Florence Baptistery. Atmos Environ 45:5979–5987CrossRefGoogle Scholar
  60. 60.
    Niklasson A, Johansson L-G, Svensson J-E (2004) Atmospheric corrosion of historical organ pipes: influence of acetic and formic acid vapour and water leaching on lead. In: Proceedings of Metal 2004 National Museum of Australia Canberra ACT 4–8 October 2004 (Canberra 2004), National Museum of Australia, pp 273–280Google Scholar
  61. 61.
    Kontozova-Deutsch V, Krata A, Deutsch F, Bencs L, Van Grieken R (2008) Efficient separation of acetate and formate by ion chromatography: application to air samples in a cultural heritage environment. Talanta 75:418–423CrossRefGoogle Scholar
  62. 62.
    Ghedini N, Sabbioni C, Bonazza A, Gobbi G (2006) Chemical–thermal quantitative methodology for carbon speciation in damage layers on building surfaces. Environ Sci Technol 40:939–944CrossRefGoogle Scholar
  63. 63.
    Hodgkins RE, Grzywacz CM, Garrell RL (2011) An improved ion chromatography method for analysis of acetic and formic acid vapours. e-Preserv Sci 8:74–80Google Scholar
  64. 64.
    Kontozova-Deutsch V, Deutsch F, Bencs L, Krata A, Van Grieken R, De Wael K (2011) Optimization of the ion chromatographic quantification of airborne fluoride, acetate and formate in the Metropolitan Museum of Art, New York. Talanta 86:372–376CrossRefGoogle Scholar
  65. 65.
    Franzoni E, Sassoni E, Graziani G (2015) Brushing, poultice or immersion? The role of the application technique on the performance of a novel hydroxyapatite-based consolidating treatment for limestone. J Cult Herit 16:173–184CrossRefGoogle Scholar
  66. 66.
    Barth HG, Boyes BE, Jackson C (1998) Size exclusion chromatography and related separation techniques. Anal Chem 70:251R–278RCrossRefGoogle Scholar
  67. 67.
    Wu CS (2003) Handbook of size exclusion chromatography and related techniques: revised and expanded. Taylor & FrancisGoogle Scholar
  68. 68.
    Berek DA (2010) Size exclusion chromatography—a blessing and a curse of science and technology of synthetic polymers. J Sep Sci 33:315–335CrossRefGoogle Scholar
  69. 69.
    Maines CA, Da la Rie ER (2005) Size-exclusion chromatography and differential scanning calorimetry of low molecular weight resins used as varnishes for paintings. Prog Org Coat 52:39–45CrossRefGoogle Scholar
  70. 70.
    Ploeger R, De la Rie ER, McGlinchey CW, Palmer M, Maines CA, Chiantore O (2014) The long-term stability of a popular heat-seal adhesive for the conservation of painted cultural objects. Polym Degrad Stab 107:307–313CrossRefGoogle Scholar
  71. 71.
    Ferreira JL, Melo MJ, Ramos AM (2010) Poly(vinyl acetate) paints in works of art: a photochemical approach. Part 1. Polym Degrad Stab 95:453–461CrossRefGoogle Scholar
  72. 72.
    Scalarone D, Chiantore O (2004) Separation techniques for the analysis of artists’ acrylic emulsion paints. J Sep Sci 27:263–274CrossRefGoogle Scholar
  73. 73.
    Somsen GW, Gooijer C, Velthors NH, Brinkman UAT (1998) Coupling of column liquid chromatography and Fourier transform infrared spectrometry. J Chromatogr A 811:1–34CrossRefGoogle Scholar
  74. 74.
    Colombini MP, Lucejko JJ, Modugno F, Orlandi M, Tolppa E-L, Zoia L (2009) A multi-analytical study of degradation of lignin in archaeological waterlogged wood. Talanta 80:61–70CrossRefGoogle Scholar
  75. 75.
    Salanti A, Zoia L, Tolppa EL, Giachi G, Orlandi M (2010) Characterization of waterlogged wood by NMR and GPC techniques. Microchem J 95:345–352CrossRefGoogle Scholar
  76. 76.
    Zoia L, Salanti A, Orlandi M (2014) Chemical characterization of archaeological wood: softwood Vasa and hardwood Riksapplet case studies. J Cult HeritGoogle Scholar
  77. 77.
    Ahn K, Hartl A, Hofmann C, Henniges U, Potthast A (2014) Investigation of the stabilization of verdigris-containing rag paper by wet chemical treatments. Herit Sci 2Google Scholar
  78. 78.
    Quye A, Littlejohn D, Pethrick RA, Stewart RA (2011) Investigation of inherent degradation in cellulose nitrate museum artefacts. Polym Degrad Stab 96:1369–1376CrossRefGoogle Scholar
  79. 79.
    Quye A, Littlejohn D, Pethrick RA, Stewart RA (2011) Accelerated ageing to study the degradation of cellulose nitrate museum artefacts. Polym Degrad Stab 96:1934–1939CrossRefGoogle Scholar
  80. 80.
    lslam AM, Phillipsl GO, Sljivo A, Snowden MJ, Williams PA (1997) A review of recent developments on the regulatory, structural and functional aspects of gum arabic. Food Hydrocoll 4:493–505Google Scholar
  81. 81.
    Kuan Y-H, Bhat R, Senan C, Williams PA, Karim AA (2009) Effects of ultraviolet irradiation on the physicochemical and functional properties of gum Arabic. J Agric Food Chem 57:9154–9159CrossRefGoogle Scholar
  82. 82.
    Duce C, Ghezzi L, Onor M, Bonaduce I, Colombini MP, Tinè MR, Bramanti M (2012) Physico-chemical characterization of protein–pigment interactions in tempera paint reconstructions: casein/cinnabar and albumin/cinnabar. Anal Bioanal Chem 402:2183–2193CrossRefGoogle Scholar
  83. 83.
    Duce C, Bramanti E, Ghezzi L, Bernazzani L, Bonaduce I, Colombini MP, Spepi A, Biagi S, Tine MR (2013) Interactions between inorganic pigments and proteinaceous binders in reference paint reconstructions. Dalton Trans 42:5945–6236CrossRefGoogle Scholar
  84. 84.
    Theodorakopoulos C, Boon JJ (2011) A high performance size exclusion chromatographic study on the depth-dependent gradient in the molecular weight of aged triterpenoid varnish films. Prog Org Coat 72:778–783CrossRefGoogle Scholar
  85. 85.
    Berg JDJ, Vermist ND, Carlyle L, Holcˇapek M, Boon JJ (2004) Effects of traditional processing methods of linseed oil on the composition of its triacylglycerols. J Sep Sci 27:181–199CrossRefGoogle Scholar
  86. 86.
    Vareckova D, Podzimek S, Lebduska J (2006) Characterization of alkyd resins by size exclusion chromatography coupled with a multi-angle light scattering photometer. Anal Chim Acta 557:31–36CrossRefGoogle Scholar

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© Springer International Publishing Switzerland 2016

Authors and Affiliations

  1. 1.Department of Chemistry and Industrial ChemistryUniversity of PisaPisaItaly

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