PDF herunterladen
Planta Med 2015; 81(07): 606-612
DOI: 10.1055/s-0035-1545873
Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Quantitation of Crocins and Picrocrocin in Saffron by HPLC: Application to Quality Control and Phytochemical Differentiation from Other Crocus Taxa

Nikolaos Stavros Koulakiotis
1   Department of Pharmacy, University of Patras, Rio, Greece
5   GAIA Research Center, Bioanalytical Department, The Goulandris Natural History Museum, Kifissia, Greece
,
Evangelos Gikas
2   Department of Pharmacy, Division of Pharmaceutical Chemistry, University of Athens, Athens, Greece
,
Gregoris Iatrou
3   Department of Biology, Division of Plant Biology, University of Patras, Rio, Greece
,
Fotini N. Lamari
1   Department of Pharmacy, University of Patras, Rio, Greece
,
Anthony Tsarbopoulos
4   Department of Pharmacology, University of Athens Medical School, Athens, Greece
5   GAIA Research Center, Bioanalytical Department, The Goulandris Natural History Museum, Kifissia, Greece
› Institutsangaben
Weitere Informationen

Publikationsverlauf

received 05. August 2014
revised 02. Februar 2015

accepted 07. Februar 2015

Publikationsdatum:
08. Mai 2015 (online)

Auch verfügbar auf
    Lizenzen und Reprints

Abstract

A chromatographic method was developed and fully validated for the determination of the major saffron constituents, i.e., picrocrocin and five major crocins. Dried samples (styles of Crocus sativus and other Crocus taxa) were extracted with MeOH : water (1 : 1, v/v), and chromatographic separation of the analytes was achieved by reversed-phase chromatography using a gradient elution. Full validation was performed using spiked samples with analytes, which were isolated, purified, and characterized by MS due to a lack of commercial standards. The method showed a good fit (r2 > 0.999) for all analytes with limit of quantitation values in the range of 1–15 µg/mL, and demonstrated adequate intra- and inter-precision (< 15 % RSD) and accuracy (< 7 % RE). The method was applied to the analysis of various commercial saffron samples and of indigenous Crocus taxa and allowed for the first time the absolute quantitation of several Crocus components.

Key words

Crocus sativus - Iridaceae - saffron - reversed-phase HPLC - quantification - validation - crocins - picrocrocin
 

  • References

  • 1 Fernandez JA, Santana O, Guardiola JL, Molina RV, Heslop-Harrison P, Borbely G, Branca F, Argento S, Maloupa E, Talou T, Thiercelin JM, Gasimov K, Vurdu H, Roldan M, Santaella M, Sanchis E, Garcia-Luis A, Suranyi G, Molnar A, Sramko G, Gulyas G, Balazs L, Horvat O, Rodriguez MF, Sanchez-Vioque R, Escolano MA, Reina JV, Krigas N, Pastor T, Renau-Morata B, Raynaud C, Ibadli O, Polissiou M, Tsimidou MZ, Tsaftaris A, Sharaf-Eldin M, Medina J, Constantinidis T, Karamplianis T, De-Los-Mozos-Pascual M. The World Saffron and Crocus collection: strategies for establishment, management, characterisation and utilisation. Genet Resour Crop Ev 2011; 58: 125-137
    CrossrefPubMedGoogle Scholar
  • 2 Bathaie SZ, Mousavi SZ. New applications and mechanisms of action of saffron and its important ingredients. Crit Rev Food Sci Nutr 2010; 50: 761-786
    CrossrefPubMedGoogle Scholar
  • 3 Chryssanthi DG, Lamari FN, Iatrou G, Pylara A, Karamanos NK, Cordopatis P. Inhibition of breast cancer cell proliferation by style constituents of different Crocus species. Anticancer Res 2007; 27: 357-362
    PubMedGoogle Scholar
  • 4 Castillo R, Fernandez JA, Gomez-Gomez L. Implications of carotenoid biosynthetic genes in apocarotenoid formation during the stigma development of Crocus sativus and its closer relatives. Plant Physiol 2005; 139: 674-689
    CrossrefPubMedGoogle Scholar
  • 5 Hill T. The contemporary encyclopedia of herbs & spices: seasonings for the global kitchen. Hoboken: John Wiley & Sons; 2004
    Google Scholar
  • 6 Schmidt M, Betti G, Hensel A. Saffron in phytotherapy: pharmacology and clinical uses. Wien Med Wochenschr 2007; 157: 315-319
    CrossrefPubMedGoogle Scholar
  • 7 International Standards Organisation. Saffron (Crocus sativus L.). Part A. Specifications, Part B. Test methods. ISO/TS3632-1,2. Zurich, Switzerland: ISO; 2003
    Google Scholar
  • 8 Lozano P, Castellar MR, Simancas MJ, Iborra JL. Quantitative high-performance liquid chromatographic method to analyse commercial saffron (Crocus sativus L.) products. J Chromatogr A 1999; 830: 477-483
    CrossrefPubMedGoogle Scholar
  • 9 Lage M, Cantrell CL. Quantification of saffron (Crocus sativus L.) metabolites crocins, picrocrocin and safranal for quality determination of the spice grown under different environmental Moroccan conditions. Sci Hortic 2009; 121: 366-373
    CrossrefPubMedGoogle Scholar
  • 10 Zougagh M, Rios A, Valcarcel M. An automated screening method for the fast, simple discrimination between natural and artificial colorants in commercial saffron products. Anal Chim Acta 2005; 535: 133-138
    CrossrefPubMedGoogle Scholar
  • 11 Anastasaki EG, Kanakis CD, Pappas C, Maggi L, Zalacain A, Carmona M, Alonso GL, Polissiou MG. Quantification of crocetin esters in saffron (Crocus sativus L.) using Raman spectroscopy and chemometrics. J Agric Food Chem 2010; 58: 6011-6017
    CrossrefPubMedGoogle Scholar
  • 12 Anastasaki E, Kanakis C, Pappas C, Maggi L, del Campo CP, Carmona M, Alonso GL, Polissiou MG. Differentiation of saffron from four countries by mid-infrared spectroscopy and multivariate analysis. Eur Food Res Technol 2010; 230: 571-577
    CrossrefPubMedGoogle Scholar
  • 13 Zalacain A, Ordoudi SA, Diaz-Plaza EM, Carmona M, Blazquez I, Tsimidou MZ, Alonso GL. Near-infrared spectroscopy in saffron quality control: determination of chemical composition and geographical origin. J Agric Food Chem 2005; 53: 9337-9341
    CrossrefPubMedGoogle Scholar
  • 14 Koulakiotis NS, Pittenauer E, Halabalaki M, Tsarbopoulos A, Allmaier G. Comparison of different tandem mass spectrometric techniques (ESI-IT, ESI- and IP-MALDI-QRTOF and vMALDI-TOF/RTOF) for the analysis of crocins and picrocrocin from the stigmas of Crocus sativus L. Rapid Commun Mass Spectrom 2012; 26: 670-678
    CrossrefPubMedGoogle Scholar
  • 15 Alonso GL, Salinas MR, Garijo J, Sanchez-Fernandez MA. Composition of crocins and picrocrocin from Spanish saffron (Crocus sativus L.). J Food Qual 2001; 24: 219-233
    CrossrefPubMedGoogle Scholar
  • 16 Gonda S, Parizsa P, Suranyi G, Gyemant G, Vasas G. Quantification of main bioactive metabolites from saffron (Crocus sativus) stigmas by a micellar electrokinetic chromatographic (MEKC) method. J Pharm Biomed Anal 2012; 66: 68-74
    CrossrefPubMedGoogle Scholar
  • 17 Tarantilis PA, Tsoupras G, Polissiou M. Determination of saffron (Crocus sativus L) components in crude plant extract using high-performance liquid-chromatography-uv-visible photodiode-array detection-mass spectrometry. J Chromatogr A 1995; 699: 107-118
    CrossrefPubMedGoogle Scholar
  • 18 VanCalsteren MR, Bissonnette MC, Cormier F, Dufresne C, Ichi T, LeBlanc JCY, Perreault D, Roewer I. Spectroscopic characterization of crocetin derivatives from Crocus sativus and Gardenia jasminoides . J Agric Food Chem 1997; 45: 1055-1061
    CrossrefPubMedGoogle Scholar
  • 19 Carmona M, Sanchez AM, Ferreres F, Zalacain A, Tomas-Barberan F, Alonso GL. Identification of the flavonoid fraction in saffron spice by LC/DAD/MS/MS: Comparative study of samples from different geographical origins. Food Chem 2007; 100: 445-450
    CrossrefPubMedGoogle Scholar
  • 20 Lech K, Witowska-Jarosz J, Jarosz M. Saffron yellow: characterization of carotenoids by high performance liquid chromatography with electrospray mass spectrometric detection. J Mass Spectrom 2009; 44: 1661-1667
    PubMedGoogle Scholar
  • 21 Verma RS, Middha D. Analysis of Saffron (Crocus sativus L. Stigma) Components by LC-MS-MS. Chromatographia 2010; 71: 117-123
    CrossrefPubMedGoogle Scholar
  • 22 Carmona M, Zalacain A, Sanchez AM, Novella JL, Alonso GL. Crocetin esters, picrocrocin and its related compounds present in Crocus sativus stigmas and Gardenia jasminoides fruits. Tentative identification of seven new compounds by LC-ESI-MS. J Agric Food Chem 2006; 54: 973-979
    CrossrefPubMedGoogle Scholar
  • 23 Tarantilis PA, Polissiou M, Manfait M. Separation of picrocrocin, cis-trans-crocins and safranal of saffron using high-performance liquid-chromatography with photodiode-array detection. J Chromatogr A 1994; 664: 55-61
    CrossrefPubMedGoogle Scholar
  • 24 Li N, Lin G, Kwan YW, Min ZD. Simultaneous quantification of five major biologically active ingredients of saffron by high-performance liquid chromatography. J Chromatogr A 1999; 849: 349-355
    CrossrefPubMedGoogle Scholar
  • 25 Caballero-Ortega H, Pereda-Miranda R, Abdullaev FI. HPLC quantification of major active components from 11 different saffron (Crocus sativus L.) sources. Food Chem 2007; 100: 1126-1131
    CrossrefPubMedGoogle Scholar
  • 26 Sanchez AM, Carmona M, Zalacain A, Carot JM, Jabaloyes JM, Alonso GL. Rapid determination of crocetin esters and picrocrocin from saffron spice (Crocus sativus L.) using UV-visible spectrophotometry for quality control. J Agric Food Chem 2008; 56: 3167-3175
    CrossrefPubMedGoogle Scholar
  • 27 Lechtenberg M, Schepmann D, Niehues M, Hellenbrand N, Wunsch B, Hensel A. Quality and functionality of saffron: quality control, species assortment and affinity of extract and isolated saffron compounds to NMDA and sigma(1) (sigma-1) receptors. Planta Med 2008; 74: 764-772
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 28 Cossignani L, Urbani E, Simonetti MS, Maurizi A, Chiesi C, Blasi F. Characterisation of secondary metabolites in saffron from central Italy (Cascia, Umbria). Food Chem 2014; 143: 446-451
    CrossrefPubMedGoogle Scholar
  • 29 Carmona M, Zalacain A, Pardo JE, Lopez E, Alvarruiz A, Alonso GL. Influence of different drying and aging conditions on saffron constituents. J Agric Food Chem 2005; 53: 3974-3979
    CrossrefPubMedGoogle Scholar
  • 30 Kyriakoudi A, Chrysanthou A, Mantzouridou F, Tsimidou MZ. Revisiting extraction of bioactive apocarotenoids from Crocus sativus L. dry stigmas (saffron). Anal Chim Acta 2012; 755: 77-85
    CrossrefPubMedGoogle Scholar
  • 31 Chen Y, Cai L, Zhao C, Xu HC, Cao CY, Liu Y, Jia L, Yin HX, Chen C, Zhang H. Spectroscopic, stability and radical-scavenging properties of a novel pigment from gardenia. Food Chem 2008; 109: 269-277
    CrossrefPubMedGoogle Scholar
  • 32 Tsimidou M, Biliaderis CG. Kinetic studies of saffron (Crocus sativus L) quality deterioration. J Agric Food Chem 1997; 45: 2890-2898
    CrossrefPubMedGoogle Scholar
  • 33 Vickackaite V, Romani A, Pannacci D, Favaro G. Photochemical and thermal degradation of a naturally occurring dye used in artistic painting. A chromatographic, spectrophotometric and fluorimetric study on saffron. Int J Photoenergy 2004; 6: 175-183
    CrossrefPubMedGoogle Scholar
  • 34 Montoro P, Maldini M, Luciani L, Tuberoso CIG, Congiu F, Pizza C. Radical scavenging activity and LC-MS metabolic profiling of petals, stamens, and flowers of Crocus sativus L. J Food Sci 2012; 77: C893-C900
    CrossrefPubMedGoogle Scholar
  • 35 Ordoudi SA, Tsimidou MZ. Saffron quality: effect of agricultural practices, processing and storage. Production practices and quality assessment of food crops, Volume 1. Dordrecht: Springer; 2004: 209-260
    CrossrefGoogle Scholar
  • 36 Harmonised Tripartite Guideline on Validation of Analytical Procedures (Q2A). International Conference on Harmonisation of Technical Requirements for Registrations of Pharmaceuticals for Human Use (ICH). Geneva: ICH; 1994
    Google Scholar
  • 37 Harmonised Tripartite Guideline on Validation of Analytical Procedures: Methodology (Q2B). International Conference on Harmonisation of Technical Requirements for Registrations of Pharmaceuticals for Human Use (ICH). Geneva: ICH; 1996
    Google Scholar