Discrimination of Saffron Based on Thin-Layer Chromatography and Image Analysis

This paper presents a new method based on thin-layer chromato-graphic separation, and image analysis (TLC-IA) of saffron colorant compounds for the discrimination of saffron from different origins. The color compounds of dried and ground saffron stigmas are extracted and separated by thin-layer chromatography at optimum conditions. After chromatographic development, the chro-matograms are photographed, and intensity profiles of RGB characteristics are produced and processed by specially designed image analysis software. The main factors such as the extraction procedure of color compounds, chromatographic system (stationary and mobile phase), and designing of image analysis software are described. This method includes a new and fast data acquisition and does not require expensive instruments. The method also allows for comparison of different types of saffron using RF values, color range, and intensity of the separated color components. This method is successfully applied to determine the saffron from different regions of Iran and various adulteration colorants.

[1]  K. Sink,et al.  Development of an improved procedure for extraction and quantitation of safranal in stigmas of Crocus sativus L. using high performance liquid chromatography , 2000 .

[2]  Ranjit Singh Verma,et al.  Analysis of Saffron (Crocus sativus L. Stigma) Components by LC–MS–MS , 2010 .

[3]  Gonzalo L. Alonso,et al.  FT-NIR SPECTROMETRY APPROACH FOR DETERMINING SAFFRON ORIGIN , 2004 .

[4]  Amaya Zalacain,et al.  Near-infrared spectroscopy in saffron quality control: determination of chemical composition and geographical origin. , 2005, Journal of agricultural and food chemistry.

[5]  M Carmona,et al.  Screening method for the detection of artificial colours in saffron using derivative UV-Vis spectrometry after precipitation of crocetin , 2005, Food additives and contaminants.

[6]  M. de la Guardia,et al.  PLS-NIR determination of total sugar, glucose, fructose and sucrose in aqueous solutions of fruit juices , 1997 .

[7]  J. Nevado,et al.  Simultaneous fluorimetric determination of pyridoxal, pyridoxamine and pyridoxic acid by partial least squares using non-linear variable angle synchronous spectra , 1998 .

[8]  Albert,et al.  Food , 1917, Nature.

[9]  C. Pappas,et al.  Geographical differentiation of saffron by GC–MS/FID and chemometrics , 2009 .

[10]  Amaya Zalacain,et al.  Analysis of saffron volatile fraction by TD–GC–MS and e-nose , 2006 .

[11]  Á. Ríos,et al.  Use of non-aqueous capillary electrophoresis for the quality control of commercial saffron samples. , 2005, Journal of chromatography. A.

[12]  D. Djozan,et al.  Forensic discrimination of blue ballpoint pen inks based on thin layer chromatography and image analysis. , 2008, Forensic science international.

[13]  Margaret Nichols.  Trans , 2015, De-centering queer theory.

[14]  Gonzalo L. Alonso,et al.  Worldwide market screening of saffron volatile composition , 2009 .

[15]  M. Cánovas,et al.  TLC preparative purification of picrocrocin, HTCC and crocin from saffron , 1992 .

[16]  A. Espinosa-Mansilla,et al.  Simultaneous determination of chlorpyrifos and carbaryl by differential degradation using diode-array spectrophotometry optimized by partial least squares , 1994 .

[17]  N. G. Kovaleva,et al.  [Medicinal plants]. , 1954, Aptechnoe delo.

[18]  Javad Feizy,et al.  LC Determination of Adulterated Saffron Prepared by Adding Styles Colored with Some Natural Colorants , 2007 .

[19]  M. Manfait,et al.  Separation of picrocrocin, cis-trans-crocins and safranal of saffron using high-performance liquid chromatography with photodiode-array detection. , 1994, Journal of chromatography. A.

[20]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[21]  C. Pappas,et al.  Differentiation of saffron from four countries by mid-infrared spectroscopy and multivariate analysis , 2010 .

[22]  Maria Z. Tsimidou,et al.  Evaluation of the colouring strength of saffron spice by UV-Vis spectrometry , 1996 .

[23]  J. Iborra,et al.  A non-destructive method to determine the safranal content of saffron (Crocus sativus L.) by supercritical carbon dioxide extraction combined with high-performance liquid chromatography and gas chromatography. , 2000, Journal of biochemical and biophysical methods.

[24]  K. Plataniotis,et al.  Color Image Processing : Methods and Applications , 2006 .

[25]  F. Cormier,et al.  In vitro formation of crocetin glucosyl esters by Crocus sativus callus extract. , 1997, Planta medica.

[26]  G. Alonso,et al.  Solid-phase extraction for picrocrocin determination in the quality control of saffron spice (Crocus sativus L.) , 2009 .

[27]  Gonzalo L. Alonso,et al.  COMPOSITION OF CROCINS AND PICROCROCIN FROM SPANISH SAFFRON (CROCUS SATIVUS L.) , 2001 .

[28]  A. Zalacain,et al.  COMPARISON OF DIFFERENT EXTRACTION PROCEDURES AND HPLC METHODS TO DETECT CROCINS IN SAFFRON , 2004 .

[29]  A. Zalacain,et al.  Identification of the flavonoid fraction in saffron spice by LC/DAD/MS/MS: Comparative study of samples from different geographical origins , 2007 .

[30]  Ioannis Pitas,et al.  Digital color restoration of old paintings , 2000, IEEE Trans. Image Process..

[31]  Charles L. Cantrell,et al.  Quantification of saffron (Crocus sativus L.) metabolites crocins, picrocrocin and safranal for quality determination of the spice grown under different environmental Moroccan conditions , 2009 .

[32]  J. Iborra,et al.  A quantitative high-performance liquid chromatographic method to analyse commercial saffron (Crocus sativus L.) products. , 1999 .

[33]  S. Garrigues,et al.  PLS-UV spectrophotometric method for the simultaneous determination of paracetamol, acetylsalicylic acid and caffeine in pharmaceutical formulations , 1997 .

[34]  G. A. Ravishankar,et al.  Methods for the analysis of the saffron metabolites crocin, crocetins, picrocrocin and safranal for the determination of the quality of the spice using thin-layer chromatography, high-performance liquid chromatography and gas chromatography , 1992 .

[35]  P. Variyar,et al.  Chemical investigation of gamma-irradiated saffron (Crocus sativus L.). , 2001, Journal of agricultural and food chemistry.

[36]  P. Tarantilis,et al.  Determination of saffron (Crocus sativus L.) components in crude plant extract using high-performance liquid chromatography-UV-visible photodiode-array detection-mass spectrometry. , 1995, Journal of chromatography. A.

[37]  G. Alonso,et al.  Crocin, safranal and picrocrocin from saffron (Crocus sativus L.) inhibit the growth of human cancer cells in vitro. , 1996, Cancer letters.

[38]  Gian Gabriele Franchi,et al.  High Performance Thin Layer Chromatographic Quantitative Analysis of Picrocrocin and Crocetin, Active Principles of Saffron (Crocus sativusL.-Iridaceae): A New Method , 1996 .

[39]  Ulf Ellervik,et al.  Evaluation of quantitative thin layer chromatography using staining reagents. , 2007, Journal of chromatography. A.

[40]  Fikart I. Abdullaev.  Cancer Chemopreventive and Tumoricidal Properties of Saffron (Crocus sativus L.) , 2002, Experimental biology and medicine.

[41]  Á. Ríos,et al.  An automated screening method for the fast, simple discrimination between natural and artificial colorants in commercial saffron products , 2005 .

[42]  M. Recio,et al.  An Update Review of Saffron and its Active Constituents , 1996 .

[43]  M. Galera,et al.  Resolution of ternary mixtures of salicylic, salicyluric and gentisic acids by partial least squares and principal component regression: Optimization of the scanning path in the excitation-emission matrices , 1995 .

[44]  G. Lin,et al.  Simultaneous quantification of five major biologically active ingredients of saffron by high-performance liquid chromatography. , 1999, Journal of chromatography. A.

[45]  M. Manfait,et al.  FT-IR, FT-Raman spectroscopic study of carotenoids from saffron (Crocus sativus L.) and some derivatives , 1998 .

[46]  Rogelio Pereda-Miranda,et al.  HPLC quantification of major active components from 11 different saffron (Crocus sativus L.) sources , 2007 .