Evaluation of Crocins in Cheeses Made with Saffron by UHPLC

Abstract

Saffron is a widespread consumed spice with potential health promoting compounds. In dairy technologies it is often used to enhance color and flavor of cheeses, so it would be recommended to know the content of saffron functional compounds in cheese made with this spice, as they could still have potential bioactivity. For this purpose, an UHPLC (ultra high performance liquid chromatography) method was developed, with the aim of reducing analysis time and solvent consumption. A methanol:water solution (80:20 v/v) was used to extract saffron compounds from cheese with stirring for 1 h in the dark at room temperature, then the samples were centrifuged at 3500 rpm, for 5 min at 4 °C and the residues were extracted twice. A linear gradient elution of acetonitrile in water allowed to simultaneously determine picrocrocin and crocins in saffron in a short time (16 min), and allowed a quantitative determination of crocins in commercial cheeses. Keywords: cheese, saffron, crocins, UHPLC IntroductionSaffron is the red dried stigmas of Crocus sativus L. flowers, now successfully cultivated in European countries (Greece, Spain, Italy, France and Switzerland) and in Morocco, Egypt, Israel, Azerbaijan, Pakistan, India, New Zealand, Australia and China.Recently, a review by Akowuah and Htar 1 reported many therapeutic properties of saffron chemical constituents, such as anticancer, cardioprotective, antidepressant, anxiolytic and anticonvulsant activities, improving learning and memory skills and management of metabolic syndrome diseases. The bioactive compounds mainly responsible of the therapeutic properties of saffron are crocins, crocetin, picrocrocin and safranal. Crocins, responsible for color and coloring properties, are a group of high water soluble cis and trans carotenoids that are sugar esters of crocetin (8,8'-diapocarotenedioic acid), with different sugar moieties, such as glucose, gentiobiose and neapolitanose. Trans-crocetin di(β-D-gentiobiosyl) ester is the most abundant crocin in saffron 2 with a high solubility being attributed to the sugar moieties. The bitter taste of saffron is derived primarily from picrocrocin (4-(β-D-glucopyranosyloxy)-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde), a monoterpene glycoside produced from zeaxanthin degradation. Crocins represent from 0.5 to 32.4% of saffron's dry matter, 3 while picrocrocin ranges from 0.8 to 26.6% on a dry matter. [3][4][5] Food items containing saffron are generally expected to provide an added value for consumers, thanks to its potential health promoting compounds. For this reason, saffron has been widely studied and many analytical methods for its compounds determination are available: in particular, the standard method, 6 which specifies the test methods for dried saffron (filaments and powder) obtained from the Crocus sativus L. flower, is based on the use of UV-Vis spectrophotometry to determine the quality of saffron in international commercial agreements, but many other analytical methods were developed, such as: thin layer chromatography, 4 hig...

Cow and Ewe Cheeses Made with Saffron: Characterization of Bioactive Compounds and Their Antiproliferative Effect in Cervical Adenocarcinoma (HeLa) and Breast Cancer (MDA-MB-231) Cells

“…Picrocrocin (peak 1 in Figure 1 a) was identified according to retention time of the standard and its UV spectrum [ 16 ], as reported in Figure 2 . However, this molecule, to which saffron taste is related, was observed only in saffron extract and not in cheese extracts, confirming previous data reported in the literature according to which the lack of picrocrocin in the hydroalcoholic extract of cheese is probably due to its degradation during cheese making [ 16 ].…”

“…Although there are many studies regarding saffron addition to cheese infused [ 11 , 12 , 13 ], very limited works are reported in the literature concerning the characterization of saffron bioactive molecules in cheese, most of them regarding the determination of the volatile profile, including safranal [ 14 , 15 ], and only one regarding crocins and picrocrocin evaluation in cheese [ 16 ]. Furthermore, the antiproliferative effects of saffron have been mainly evaluated in pure saffron components [ 17 , 18 , 19 , 20 ] or its extracts [ 21 , 22 , 23 ], while there is a lack of studies concerning evaluation of the antiproliferative effect of saffron extract within a food matrix or in a complex mixture, additionally, no evaluation of the influence of the food chemical environment on saffron’s antiproliferative action has been reported.…”

A Review of the Preservation of Hard and Semi-Hard Cheeses: Quality and Safety

“…The AA and antimicrobial properties of saffron cheeses have been described, but the number of different cheeses made with saffron is currently very scarce. Thus, it would be of interest to know the contents of potential health-promoting compounds that can be transferred from saffron to the cheese matrix [ 120 , 121 , 122 ]. Different concentrations of saffron were added (concentrations were not indicated) to pressed pasteurized sheep milk cheeses ripened for 180 days.…”

Natural Preservatives from Plant in Cheese Making

“…Different scientific works also reported the addition of saffron to cheese [83][84][85] to generally enhance color and flavor of the dairy products [86]. Color and coloring properties of saffron are related to crocins [86], sugar esters of crocetin, while saffron aroma is mainly due to safranal [84], the degradation product of picrocrocin, which is responsible for the bitter taste of saffron [86]. Due to the presence of all these analytes, saffron is also considered a source of bioactive compounds [86].…”

“…Color and coloring properties of saffron are related to crocins [86], sugar esters of crocetin, while saffron aroma is mainly due to safranal [84], the degradation product of picrocrocin, which is responsible for the bitter taste of saffron [86]. Due to the presence of all these analytes, saffron is also considered a source of bioactive compounds [86]. Significant differences were observed in flavor among control cheese and cheeses added with different saffron concentrations, but as ripening time increased, these differences were less evident [83,85].…”