Saffron is the commercial name for the dried red stigmas of the Crocus sativus L. flower. It is appreciated for adding color, flavor, and a particular aroma to different food dishes or drinks (paella in Spain, Milanese risotto in Italy, lussekatter buns in Sweden, and alcoholic beverages). It is considered a high-priced condiment (1500–2200 euro/kg) due to the considerable labor involved in its production since it requires manual harvesting as well as a laborious handling process (sorting, drying, and storage). Saffron’s principal producers are Iran and Spain, whereas the leading importers are Spain, Hong Kong, and the United States. Saffron’s quality is essential for consumers in the food industry and is based on the concentration of its apocarotenoids and their respective sensory attributes: crocin’s coloring strength, picrocrocin’s bitter taste, and safranal’s aromatic intensity
1. Saffron’s Chemical Composition
Saffron contains more than 150 compounds (volatile and non-volatile) including carotenoids (crocetin, crocin, β-carotene, lycopene, and zeaxanthin), monoterpene aldehydes (picrocrocin and safranal), monoterpenoids, and isopherones
[1][2]. However, it also contains other compounds such as flavonoids, vitamins, proteins, and amino acids
[3]. Saffron owes its sensory and functional properties mainly to the presence of its carotenoid derivatives, synthesized throughout flowering but also during the whole production process
[4]. These compounds include crocin, crocetin, picrocrocin, and safranal, which are the secondary or bioactive metabolites
[1][4][5][6]. Saffron’s quality depends on its chemical profile and is directly related to the geographic area, climate variability, environmental practices, genetic traits, soil composition, cultivation conditions, and processing and storage methods
[7][8]. Nevertheless, according to the ISO standards (3632-1:2011 and ISO 3632-2:2010), the value and quality of the stigma are measured based on the content of the color components (crocin and crocetin), the bitter taste component (picrocrocin), and the volatile compounds responsible for the odor and aroma (safranal). These specific parameters are influenced by the environmental conditions, extraction method, purification, etc.
[2][9][10][11][12]. Some studies have been conducted on the extraction of bioactive compounds from saffron using the concept of green chemistry
[13]. Some research on saffron stability demonstrates that temperature and humidity exert a strong influence on the degradation of the principal active ingredients
[1].
1.1. Saffron’s Important Apocarotenoids
Crocin: The main bioactive compound of saffron was isolated by Aschoff in 1818, reporting a family of yellowish-red water-soluble carotenoids (mono-glycosyl or di-glycosyl-polyene esters) of 20 carbons
[1][13][14][15][16]. In other words, this was a group of compounds formed by crocetin esterification (dicarboxylic carotenoid), which were classified according to their sugar fractions
[15]. The cis/trans-X-R1R2 crocin abbreviation system is used based on three main characteristics: (a) cis/trans isomers, (b) X: number of glucose components (1–5), and (c) type of structure in R1 and R2 (acid form: H; glucose: g; gentiobiose: G; Neapolitan: n; or triglucose: t.) (Suchareau et al. (2021)). The most represented crocins are trans-4-GG, trans-3-Gg, trans-2-G, trans-2-gg, trans-5-tG, and trans-1-g, among others
[15][17][18][19][20][21][22][23][24][25][26][27].
Crocins are unusual apocarotenoids since their terminal glycoside rings confer high solubility. These pigments are detected in the red lobes of the stigmas of the
Crocus sativus flower
[9][17] and their content is proportional to the color and quality index. However, it should be noted that zeaxanthin (fat-soluble carotenoid) can also influence the color
[28]. Crocins as such have low stability and lose their functionality during exposure to heat, oxygen, light absorption, acidic environments, and/or due to the presence of additives
[4]. Therefore, the drying and storage temperatures are important for proper color development
[25]; poor storage conditions lead to color pigment degradation
[29]. Several factors are related to the concentration of these pigments in saffron stigmas, which are mainly the geographical growing region, crop conditions, type of soil, plant genetic traits, climate, planting time (rate), seed/crown rate, planting depth, corm size/weight, crop density, nutrient management, weed management, growth regulators, harvest and postharvest management, and drying conditions
[30][31]. Finally, crocin (digentiobiose ester of crocetin) is recognized as a natural food-grade dye that displays biological activity such as antigenotoxic, cytotoxic, antioxidant, anti-inflammatory, anti-atherosclerotic, anti-diabetic, hypotensive, hypolipidemic, hypoglycemic, and antidepressant properties
[2][9][10].
Crocetins are lipophilic carotenoids derived from the hydrolysis of crocin glycosides, which is a crocin aglycone
[24]. It contains a carboxyl group at each end of the polyene chain
[17]; these groups of compounds (α-crocetin or crocetin I, crocetin II, β-crocetin, γ-crocetin) are produced from the degradation of zeaxanthin
[32].
Picrocrocin’s structure was established by Khun and Winterstein in 1934
[16]. It is a colorless and odorless glycoside monoterpene (4-hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde or hydroxy-β-cyclocitral: HTCC and glucose), a product of the degradation of zeaxanthin, and is responsible for saffron’s bitter taste
[1][2][6][13][14][33]. Picrocrocin is the second most abundant component in dry matter content
[23][32][34]. During the drying process (35–50 °C for 4–7 h), picrocrocin’s temperature and/or hydrolysis form an aglycone
[32][35]. Therefore, picrocrocin decreases during dehydration, whereas safranal is absent before drying
[36].
Safranal is an aldehyde monoterpene and the volatile component responsible for saffron essential oil. HTCC (hydroxy-β-cyclocitral or 4-hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde) is regarded by many authors as a safranal precursor. This compound is obtained by chemical or enzymatic hydrolysis (dissociation) or when the vegetal material is dehydrated and transformed into safranal, but this also happens due to the handling and storage processes
[1][7][13][20][34][37][38]. The safranal content changes according to the duration and intensity of drying, causing quality fluctuations
[14], whereas its concentration increases with the storage and timely harvesting of flowers. However, heat and sunlight decrease the final quality and price
[2].
1.2. Hypotheses on the Method of Obtaining Apocarotenoids
There are various hypotheses on the method of obtaining these important apocarotenoids from saffron. The first theory focuses on synthesizing these compounds in the plant from protocrocin (glycosyl derivative of zeaxanthin), the substrate of an oxidative enzyme that produces a molecule of crocin and two molecules of picrocrocin. Regarding safranal, it has been described that only a minimal concentration is detected in the fresh spice
[39]. Fallahi et al.
[40] described another pathway wherein apocarotenoids, which are commercially important, are obtained by the cleavage of carotenoids (zeaxanthin and β-carotene) by the carotenoid dioxygenase enzyme, giving rise to crocetin and hydroxy-β-cyclocitral as products. Later, they propose a glycosylation (glycosyltransferases) step, which produces crocins and picrocrocin, respectively. Finally, they describe that picrocrocin is hydrolyzed to form safranal. This hypothesis is consistent with that described by Sereshti et al.
[41], who also describe other, more specific enzymes and substrates, as seen in
Figure 1.
Figure 1. Possible pathways of commercial apocarotenoids in saffron.
Possible pathways of commercial apocarotenoids in saffron.
The enzyme dioxygenase performs a 7–8C and 7′–8′C symmetric cleavage on the carotenoid zexanthin, converting it to 3-hydroxy-𝛽-cyclocitral and dialdehyde crocetin. Crocetin dialdehyde undergoes oxidation by aldehyde dehydrogenase to crocetin. Crocetin further undergoes glycosylation at the carboxyl group by the enzyme UDP-glucuronosyl transferase, forming crocin. Picrocrocetin is obtained from 3-hydroxy-𝛽-cyclocitral by glycosylation at the hydroxyl group by the enzyme UDP-glucuronosyl transferases. Picrocrocin is converted to safranal by the action of the enzyme 𝛽-glucosidase along with heat during drying
[9].
2. Saffron Quality: Compounds Related to Color, Odor, and Flavor
Saffron’s quality depends on its chemical profile, which provides the bitter taste, desirable aroma, and attractive yellowish-red color of this spice
[42][43]. Several studies on saffron stability are related to temperature, humidity, pH, light, oxygen
[35], geographical growth location, and drying and storage conditions
[44]. Since 1980, a standard quality procedure has been employed for saffron classification according to the International Standard Organization (ISO/TS 3632), which was updated in subsequent years (2003, 2010, 2011). This regulation allows saffron to be classified into distinct categories based on physical and chemical criteria: Category I—high quality; Category II—±medium quality; and Category III—low quality
[18][45][46]. The grouping parameters used are moisture content, flower residues, foreign material, ash, and coloring power. However, external parameters, such as the absence of other plants, biological micro-flora, and pesticide residues, are also used. The methodology to determine saffron’s quality using these regulations is the spectrophotometric quantification of the stigmas’ aqueous extracts (1%) at three maximum wavelengths, namely 257 nm to indicate flavor strength (picrocrocin), 330 nm related to aroma (safranal), and 440 nm for coloring force (crocins), using a 1 cm pathway quartz cell
[46][47][48][49][50]. The results are reported according to Equation (1):