Crocin and crocetin are two bioactive compounds found in saffron and gardenia. In the food industry, due to their natural yellow or orange color, they are used as food colouring. The study was carried out to optimize the high-performance liquid chromatography combined with a photodiode detector (HPLC-PDA) to simultaneously determine the crocin and crocetin content in food. Analytical samples were ultrasonicated with ACN:H2O (1:1, v/v) solvent for 30 minutes. The sample solution was determined by HPLC – PDA with the following conditions: Sunfire C18 column (250 mm x 4.6 mm; 5 µm) mobile phase consisting of 0.1% formic acid and ACN using gradient program. The method has good specificity, the standard curves of crocin and crocetin have correlation coefficients R2 > 0.999, respectively, repeatability and recovery meet AOAC requirements. The limit of detection (LOD) for crocin and crocetin were 0.5 mg/kg and 0.25 mg/kg, respectively. The method has been applied to simultaneously determine crocin and crocetin in 15 real samples and results in 13 out of 15 samples detecting crocin content, 11 out of 15 samples detecting crocetin content, and in 2 out out of 15 samples. neither crocin nor crocetin was detected. The gardenia powder group contained the highest content of crocin and crocetin (23.5 g/100g – 24.4 g/100g) and dried lychee samples - sun-dried or dried gardenia contains the second highest concentrations of crocin and crocetin in the food group.
Crocin, crocetin, high-performance liquid chromatography, HPLC, PDA, food
[1]. L. Xi, & Z. Qian, Pharmacological properties of crocetin and crocin (digentiobiosyl ester of crocetin) from saffron. Natural Product Communications, vol. 1, no. 1, 1934578X0600100112, 2006.
[2]. E. Christodoulou, M. E. Grafakou, E. Skaltsa, N. Kadoglou, N. Kostomitsopoulos, & G. Valsami, “Preparation, Chemical characterization and determination of crocetin's pharmacokinetics after oral and intravenous administration of saffron (Crocus sativus L.) aqueous extract to C57/BL6J mice," Journal of pharmacy and pharmacology, vol. 71, no. 5, pp. 753-764, 2019.
[3]. N. Ozeki, H. Oka, Y. Ito, E. Ueno, T. Goto, T. Hayashi & H. Matsumoto,"A reversedphase thin-layer chromatography/scanning densitometric method for the analysis of gardenia yellow in food using crocetin as an indicator," Journal of liquid chromatography & related technologies, vol. 24, no. 18, pp. 2849-2860, 2001.
[4]. T. Watanabe, “Separation and determination of yellow Gardenia pigments for food and iridoid constituents in Gardenia fruits by micellar electrokinetic chromatography,” Food Science and Technology International, Tokyo, vol. 4, no. 1, 54-58, 1998.
[5]. A. Girme, S. Pawar, C. Ghule, S. Shengule, G. Saste, A. K. Balasubramaniam & L. Hingorani, “Bioanalytical method development and validation study of neuroprotective extract of Kashmiri saffron using ultra-fast liquid chromatographytandem mass spectrometry (UFLC-MS/MS): In Vivo pharmacokinetics of apocarotenoids and carotenoids,” Molecules, vol. 26, no. 6, 1815, 2021.
[6]. M. Suchareau, A. Bordes, & L. Lemée, “Improved quantification method of crocins in saffron extract using HPLC-DAD after qualification by HPLC-DAD-MS,” Food Chemistry, vol. 362, 130199, 2021.
[7]. W. Yang, J. Wang, X. Li & Z. Du, "A new method research for determination of natural pigment crocin yellow in foods by solid-phase extraction ultrahigh pressure liquid chromatography," Journal of Chromatography A, vol. 1218(11), pp. 1423-1428, 2011.
[8]. W. E. Zhou, Y. Zhang, Y. Li, Y. Ling, H. N. Li, S. H. Li, & F. Zhang, "Determination of gardenia yellow colorants in soft drink, pastry, instant noodles with ultrasoundassisted xtraction by high performance liquid chromatography-electrospray ionization tandem mass spectrum," Journal of Chromatography A, vol.1446, pp. 59- 69, 2016.
[9]. N. Li, G. Lin, Y. W. Kwan, & Z. D. Min, "Simultaneous quantification of five major biologically active ingredients of saffron by high-performance liquid chromatography," Journal of Chromatography A, vol. 849, no. 2, 3pp. 49-355. 1999.
[10]. F. Yin, X. Wu, L. Li, Y. Chen, T. Lu, W. Li & W. Yin, "Quality control of Gardeniae Fructus by HPLC-PDA fingerprint coupled with chemometric methods," Journal of Chromatographic Science, vol. 53, no. 10, pp. 1685-1694, 2015.