Chemical Composition of Saffron: a Review Syed Muzaffar*A, Tanveer A

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Review article Chemical Composition of Saffron: A Review Syed Muzaffar*a, Tanveer A. Sofib and Khaliquz Zaman Khana Department of Chemistry, University of Kashmir, Srinagar 190006, India, 2Department of Zoology, University of Kashmir, Srinagar 190006, India

A R T I C L E I N F O A B S T R A C T

Keywords: Abstract: Saffron, considered being the king of spices with the costliest tag in terms of weight Saffron has not received much scientific attention within the state of Jammu and Kashmir, though this Jammu and Kashmir Cultivation is an important privileged asset to this state and constitutes approximately 16% of agricultural Chemistry income. This golden crop of our state is world famous due to its glorifying dark reddish aromatic stigmas and there are thousands of people involved in cultivation and trade of saffron within and outside Jammu and Kashmir. More importantly, there is a deep interest growing globally among scientists, herbal and pharmaceutical industries to explore biochemical and biological profile of saffron spice so that various products could be formulated based on saffron integration. Further, there was not much literature available about saffron native to Jammu and Kashmir particularly related to the impact of different drying methods and soil constituents on its chemical profile as well as nutritional values and evaluation of its activities such as anticancer, antibacterial and antifungal. Hence, these dimensions were attempted in the present study.

INTRODUCTION Saffron (Crocus sativus L.) has a significant place as a spice and 2667 hectares with an annual production of 5.61 tons (Husaini et has been an important ingredient of unani and ayurvedic al., 2013). In addition among all Indian varieties, saffron grown in formulations for the last 3500 years (Plessner et al., 1989). Jammu and Kashmir is considered to be of good quality (Behazad According to Spanish Food Code, saffron spice is defined as Crocus et al., 1992).Saffron spice has been used in seasoning, medicine, sativus L. stigmas with a maximum tolerance of styles and floral cosmetics, perfumes, dyes, in religious rites and celebrations for waste specified for various qualities in corresponding regulations over three millennia (Ulbricht et al., 2011; Perez, 1995). Egyptians (Delgado et al., 2006). Saffron belongs to the family irridaceae and and Hebrews used it to carry out ablutions in temples and sacred is a herbaceous perennial cormous plant having a height of 10 to places (Capel and Girbes, 1988). In India it is used in religious 25 cm. This plant grows on arid or semi-arid lands adaptable to ceremonies and as a sign of good omen is applied on the forehead temperate and sub-tropical climates at an altitude ranging (Madan et al., 1965; Alarcon and Sanchez, 1968). In addition to its between 600-2800 m from sea level (Agayev, 2003; Poma and culinary use, saffron spice also shows medicinal properties. It is Fontechio, 2012). Saffron is majorly being cultivated in Iran, India, known from antiquity for its antispasmodic and sedative activities Spain, Greece and Italy, however, it is also grown in other countries and has been used as a stomach stimulant (Sampathu et al., 1984). like Morocoo, France, Switzerland, Turkey, Israel, Azerbaijan, During the last decade, investigations have been focused on the Pakistan, China, Egypt, UAE, Japan, Afghanistan, Iraq, Tasmania tumoricidal and anticarcinogenic properties of saffron Australia and Mexico with minimal production (Fernández, 2007; (Abduyaev, 2002; Fernández, 2004; Fernández, 2006), and more Amir et al., 2010; Abdullaev, 2002). Iran with more than 47,000 recently on its free radical scavenging and detoxifying capacities hectares of land under saffron cultivation shares 80% of the (Ajam et al., 2010; Ordoudi et al., 2009). All these properties have world's total production (Ali, 1979; Nehvi et al., 2008). In India it been attributed to the stigmas, the sole commercially valuable is grown in the states of Jammu and Kashmir, Himachal Pradesh part of the plant whereas other parts of the plant have been much and Uttrakhand. In Jammu and Kashmir saffron has been less studied. Extracts of saffron petal have shown antidepressant cultivated since 750 AD and the total area under its cultivation is effects (Moshiri et al., 2006), antinociceptive and anti- inflammatory properties (Hosseinzadeh et al., 2002), free radical scavenging and antityrosinase activities (Li et al., 2002). * Corresponding Author : Syed Muzaffar [email protected] Chemical analysis has shown the presence of numerous components in C. sativus stigmas (Bathaie and Mousavi, 2010). These are characterized by the presence of sugars, minerals, fats,

vitamins and secondary metabolites including terpenes, flavonoids, isophorone, 2,2,6-trimethyl-1,4-cyclohexanedione, 4- anthocyanins and carotenoids. Between them, carotenoids are the ketoisophorone, 2-hydroxy-4,4,6-trimethyl-2,5-cyclohexadien-1- most important molecules because they determine color and taste of one as well as 2,6,6-trimethyl-1,4-cyclohexadiene-1- the saffron spice (Gismondi et al., 2012). Saffron's name is derived carboxaldehyde (Maggi et al., 2009) but more than 160 additional from the Arabic word for yellow, a name reflecting the high volatile components have been identified (Carmona et al., 2007). Of concentration of carotenoid pigments present in the saffron stigmas these, safranal represents approximately 30 to 70% of essential oil which contribute most to the color profile of this spice. Both and 0.001 to 0.006% of dry matter (Carmona et al., 2007; Maggi et al., lipophilic carotenoids and hydrophilic carotenoids have been 2009). Besides its typical spicy aromatic note, safranal has also been identified in saffron (Alonso et al., 2001). The lipophilic carotenoids, shown to have high antioxidant potential (Assimopoulou et al., 2005; lycopene, α-, and β- carotene and zeaxanthin have been reported in Kanakis et al., 2007) as well as cytotoxicity towards certain cancer trace amounts (Sampathu et al., 1984). However the hydrophilic cells in vitro (Melnyk et al., 2010; Escribano et al., 1996). crocins constitutes approximately 6 to16% of saffron's total dry matter depending upon the variety, growing conditions, and Isolation and characterization of carotenoids is not an easy processing methods (Gregory et al., 2005). Carotenoids are process, because they are very sensitive to light, humidity and high terpenoids and ubiquitous in nature which can be synthesized in temperatures. There have been many attempts to study their vivo through two different pathways: 1) mevalonic acid (MVA) extraction, focusing on the solvent variable. Some analytical pathway in the cytoplasm and 2) non-mevalonic acid pathway (2-C- methods have been developed to separate, identify and quantify the methyl-D-erythritol 4-phosphate pathway: MEP) in plastids that crocins. Techniques such as thin-layer chromatography (TLC) is provides the precursors for carotenoids. The MVA pathway starts commonly used in ISO normative, but the high performance liquid with synthesis of mevalonate through three molecules of acetyl CoA chromatography (HPLC) is the most effective technique for the and then continues with production of isopentenyl diphosphate analysis of crocins and related compounds in saffron extracts (IPP) molecules, geranyl geranyl pyrophosphate (GGPP), colour less (Zareena et al., 2001; Tarantilis et al., 1994; Basker and Negbi, 1985). phytoene, coloured lycopene, β-carotene and zeaxanthin (Namin, 2009; Bolhassani et al., 2014). This pathway possesses many Cold percolation method is a slow process with slightly lower enzymes which catalyzed the reactions and coded by related key yields of extraction, but the safranal content of oleoresin was found genes such as PSY, LYC, CCD, BCH and ZCD (Bolhassani et al., 2014). β- to remain more intact in this method. Solid phase extraction (SPE) is carotene with two rings is built up via cyclization of lycopene with one of the most common and least expensive purification techniques lycopene-β-cyclase (LYC). The hydroxylation of β-carotene in MVA and is considered as a convenient approach for sample preparation pathway is catalyzed by β-carotenoid hydroxylase coded by BCH in food analysis. SPE has been applied for the detection of gene to yield zeaxanthin. The biogenesis of the colour and odour adulterations by artificial colourants in saffron (ISO, 2003) and active compounds of saffron are derived through bio-oxidative purification of its components (Alonso et al., 1990). Safranal being cleavage of zeaxanthin at the points 7, 8 (7′, 8′) by zeaxanthin volatile, needs careful handling during extraction. Various extraction cleavage dioxygenase to produce crocetin dialdehyde and methods have been used for the extraction of volatiles from plant picrocrocin. In C. sativus stigmas, the final step involves material such as micro simultaneous hydro-distillation extraction glucosylation of the generated zeaxanthin cleavage products by (Iborra et al., 1992a), supercritical fluid extraction (Iborra et al., glucosyl transferase enzyme in chromoplast of stigmas (Pfander and 1992b), thermal desorption (Orfanou and Tsimidou, 1996), vacuum S c h u r t e n b e r g e r, 1 9 8 2 ; B o l h a s s a n i e t a l . , 2 0 1 4 ) . headspace (Tsimidou and Biliaderis, 1997), liquid extraction with organic solvents (Selim et al., 2000) and the ultrasound-assisted Crocin 1 (or α-crocin), a digentiobioside, is the most abundant extraction (USAE) (Lourdes et al., 2010). With the advancement in crocin with a high solubility being attributed to the sugar moieties. technology, quantification and estimation of bioactive compounds in Crocin (C44H64O24), typically deep red in color, quickly dissolves in saffron is being done with more accuracy through latest methods water to form an orange colored solution thereby making crocin like micellar electrokinetic chromatographic (MEKC) method widely used as a natural food colourant. In addition to being an (Liakopoulou-Kyriakides and Skubas, 1990). But still most of the excellent colourant, crocin also acts as an antioxidant by quenching estimation work is done by HPLC because of its accuracy and free radicals, protecting cells and tissues against oxidation reproducibility. The volatile components in dried stigma tissues are (Assimopoulou et al., 2005; Papandreou et al., 2006; Soeda et al., very important indicators for determination of quality of original 2007; Melnyk et al., 2010). The actual taste of saffron is derived saffron. There are different techniques which have been used for primarily from picrocrocin (C16H26O7) which is the second most elucidation of volatile saffron stigma composition, however, GC-MS abundant component (by weight), accounting for approximately 1% is considered to be the best choice because of its rapidness, high to 13% of saffron's dry matter (Alonso et al., 2001). Natural de- sensitivity, low limit of detection and rapid compound identification glycosylation of picrocrocin yields another important chemical using their mass spectra (Esmaeilian et al., 2012). component, safranal (C10H14O), which is mainly responsible for the aroma of saffron. Dehydration is not only important to the Differences among production areas regarding crop, processing preservation of saffron but is actually critical in the release of and classification result in geographical origin distinction of saffron safranal from picrocrocin via enzymatic activity, the reaction spice based on physicochemical and sensory properties (Carmona et yielding D-glucose and safranal, the latter being the volatile oil in al., 2006). Further from the evidence available there is also much saffron. The six major volatile compounds in saffron are safranal, uncertainty about the ideal preservation methods for saffron. The 6912 Syed Muzaffar et al. /Int J Biol Med Res.10(4):6910 - 6919

commonly used drying method of preservation is to reduce the minimal production (Fernández, 2007; Amir et al., 2010; Abdullaev, moisture content to 10-12%. This is achieved through toasting (Iran 2002). Its flower is made up of six violet tepals, three yellow stamens and Spain), air drying at room temperature and sun-drying (India) and a single pistil whose stigma, defined by three red filaments, is the (Maghsoodi et al., 2012). However saffron aromatic fingerprint has source of the saffron (Gresta, 2007). Saffron is cultivated almost an immense effect of drying methods used. Maintaing proper level of exclusively for its stigma, which once dried, forms saffron, the most saffron moisture content is an essential requirement for saffron expensive spice in the world (Sánchez-Vioquea et al., 2012). The quality in international market (Maghsoodi et al., 2012). The value taxonomic classification of saffron is as follows (Goldblatt, 1990). and quality of saffron is determined by its chemical composition (ISO 3632–1, 3631–2, 2003). The disparity in saffron drying methods of each country and region leads to loss of major components. The traditional methods of saffron drying mainly used in Jammu and Kashmir is sun drying which takes 3-5 days and occasional shade drying till moisture content is reduced to 10-12 % (Carmona et al., 2007). Hence, there is a potential scope for the designing of drying method with minimal effect on chemical composition of saffron.

The definition of quality of saffron that Dioscorides gave in the first century of our age is “The most perfect saffron for use in medicine is fresh, with good colour, a bit of white in the filaments, long, whole, difficult to crumble, not oily, full, staining ones hands when wet, neither smelling like mould nor touched by woodworm Saffron (Crocus sativus) and lastly giving off a sharp, smooth scent”. ISO (the international Kingdom : Plantae organization for standardization) is a worldwide federation of Division : Mangoliopyta national standard bodies (ISO member bodies), they have set a Class : Liliopsida classification of saffron based on minimum requirements of each Order : Asparagales quality, and this is called ISO 3632. The major focus of this measure is Family : Iridaceae to establish whether the product meets the ISO standard Genus : Crocus parameters, to get an indication of the range of quality being Species : sativus produced under different geographical locations. This classifies Bionomical name : Crocus sativus L. saffron into three categories' on the basis of large number of physical and chemical parameters such as microscopic characteristics, Synonyms presence of flowers waste, moisture and volatile matter content, ash Hindi-kesar, zaffran; Sanskrit-avarakta, saurab, mangalya, content, E-440 nm (coloring strength, E-330nm (related to safranal agnishikha, kumkuma, mangal, kusrunam; English- saffron; Arab content), E-257nm (related to picrocrocin content) etc. However, and Persian zafrah, zipharana; Guj-keshar; Tam. and Mal.- only coloring strength representing the crocetin ester content has kunkumappu; and Ger.-safran (Corradi and Micheli, 1979). traditionally been of importance for the companies which trade with saffron. This standard is however not capable of discriminating Saffron chemistry between qualities with regards to volatiles (E-330nm) or picrocrocin (E-257 nm) content. When samples fulfill the Crocus sativus stigmas are characterized by the presence of requirements of any category regarding coloring strength sugars, minerals, fats, vitamins and secondary metabolites such as (absorbance at 440nm) it also fulfills the other spectrometric terpenes, flavonoids, anthocyanins and carotenoids (Gismondi et al., parameters (E-330nm, E-257nm) for the same category (ISO 2012). Range of all chemical constituents can vary greatly due to 3632–1, 3631–2, 2003). growing conditions and country of origin.

Cultivation and taxonomic classification of saffron Proximate composition

Crocus sativus Linn belongs to family Iridaceae is a flowering Proximate composition of dried stigmas of saffron has been plant and is commonly known as saffron. It might have come from studied by Sampathu et al. 1984 (Table 1). Where, total vitamin Asia Minor, Mesopotamia or Egypt and could have evolved from one content of different samples of saffron was 3.4–5.6 µg/g and of or more predecessors. One of them was probably Crocus thiamine 0.3–0.4 µg/g. Percentage composition of saffron ash cartwrightianus Herbert (Delgado et al., 2006). Saffron is a perennial (34.46%), K2O (8.56%), Na2O (10.01%), P2O5 (7.12%), SO3 herbaceous, 10 to 25 cm high plant that develops from corms. (2.89%) and quantities of Magnesium (~565 mg/100 g), calcium Saffron is currently being cultivated in Iran, India, Spain, Greece and (~150 mg/100 g), iron (25.4– 76.5 mg/100 g), manganese (0.24–5.7 Italy, however it also grows in other countries like Morocoo, France, mg/100 g), copper (1.5–27.7 mg/100 g), zinc (0.5–15.2 mg/100 g) Switzerland, Turkey, Israel, Azerbaijan, Pakistan, China, Egypt, UAE, has been reported (Sampathu et al., 1984; Tarantilis et al., 1990; Japan, Afghanistan, Iraq, Tasmania Australia and Mexico with Alonso et al., 1998). Syed Muzaffar et al. /Int J Biol Med Res.10(4):6910 - 6919 6913

Table 1: Proximate composition of dried stigmas of saffron Chemistry in terms of quality

The main important active constituents which have been used as markers for quality of saffron and its pharmacological studies are cis and trans-crocin, crocetin, picrocrocin and safranal. Further the quality and consequently the commercial value of saffron is based on the estimation of colouring power, the bitter taste and the aroma.

Crocetin

Liakopulou et al. (1999) revealed that crocetin esters such as all- trans-crocetin di-(β-D-gentiobiosyl) or α-crocin, all-trans-crocetin β-D-gentiobiosyl-β-D-glucosyl ester, all-trans-crocetin di-(β- D- glucosyl) ester, all-trans-crocetin mono-(β-D-gentiobiosyl) ester, 13-cis-crocetin β-D-gentiobiosyl- β-D-glucosyl ester are commonly found in saffron extract and they are known as crocins. Among the glycosyl ester of crocetin, the β-crocin, digentiobiosyl ester of the crocetin (Table 2), is responsible for 10% of saffron's mass and its distinctly bright yellow orange colour. Crocetin is reported to occur Secondary metabolites to a limited extent freely in the spice. Côté et al. (2000) have reported Apart from the primary metabolites such as carbohydrates, on the properties of a glucosyltransferase which is involved in the minerals, fats and vitamins, the crocus sativus L. contains four major glucosylation of crocetin. This enzyme (uridine-5′- bioactive compounds viz., crocin (Mono-glycosyl polyene esters), diphosphoglucose- crocetin 8, 8′-glucosyltransferase), which crocetin (a natural carotenoid dicarboxylic acid precursor of crocin), glucosylates the carboxylic ends of crocetin, has been isolated from picrocrocin (monoterpene glycoside precursor of safranal and cell cultures of saffron. The glucosylation of crocetin into crocin by product of xeaxanthin degradation) and safranal, all contributing to UDP-glucose takes place according to the following sequence: colour, taste and aroma respectively (Melnyk et al., 2010) (Fig. 1). monoglucosyl ester; monogentiobiosyl and diglucosyl esters; According to Sobolev et al. (2014) presence of biologically active gentiobiosylglucosyl ester and crocin. compounds such as crocetin, picrocrocin and safranal makes this Table 2. Crocetin and Crocin derivatives of saffron spice a promising candidate for being a functional food. (LiakopoulouKyriakides and Kyriakidis, 2002).

Crocins

Water-soluble crocins are mainly responsible for colouring properties of Saffron. Various investigators have reported the structures of major crocins. Tarantilis et al. (1995) used HPLC-UV- visible photodiode-array detection on-line with mass spectrometry for the analysis of crocetin glycosides (crocins), carrying one to five glucose molecules, and differentiated their trans and cis isomers. Investigators revealed that on acid hydrolysis in absence of air, crocins yield crocetin and glucose, while hydrolysis with alcoholic ammonia results in crocetin and gentiobiose.

The antioxidant properties of crocin have also been investigated (Pham et al., 2000). Weber and Grosch (1976) have reported a carbonyl compound as a product of crocin bleaching during co- oxidation with linoleic acid by a soybean lipoxygenase. The levels of Figure 1. Major components of saffron 6914 Syed Muzaffar et al. /Int J Biol Med Res.10(4):6910 - 6919

each pigment in saffron vary due to the different origin of the plant and the overall processing conditions and storage length. Pfander and Rychener (1982) found that crocin-1 represents the 40–45% of the aqueous extract of saffron, followed by the crocetin-(β-D- gentiobiosyl)-(β-D-glucosyl) ester (35%), the crocetindi-(β-D- glucosyl) ester, as well as the crocetin-mono-(β-D-gentiobiosyl) and mono-(β-D-glucosyl) esters. On the other hand, Alonso et al.(2001) examined the content of Spanish, Indian and Iranian saffron in crocin derivatives and gave results for trans- and cis-crocins (trans-crocin: 0.46–12.12%; cis-crocin: 0.04–8.53%; trans-(β-D-gentiobiosyl)-(β- D-glucosyl) ester: 0.01–9.44%; cis-(β-D-gentiobiosyl)-(β-D- glucosyl) ester: 0.01–2.26%).

Pigments Figure 2. Production of safranal (Bolhassani et al., 2014)

Numerous C20 carotenoid pigments have been isolated from the The levels of safranal and those of the other aroma compounds dried stigmas of saffron. The main pigments are the glycosidic forms from saffron vary depending mainly upon the conditions of of crocetin (8,8′-diapocarotene-8,8′-dioic acid), D-glucose and D processing, storage and the methods of analysis of saffron. In any gentiobiose as sugar moieties, known as crocins. α-Carotene, β- case, safranal is the most abundant volatile component in the carotene, lycopene and zeaxanthin are also present in trace amounts stigmas of saffron (> 60% of essential oil) though even lower levels and mangicrocin, a xanthone-carotenoid glycosidic conjugate has have also been reported (Bolhassani et al., 2014). Some minor also been identified. The structure of dimethyl crocetin volatile compounds are believed to have been derived from non- (C11H14O2)2, a purple compound, prepared by alkaline hydrolysis volatile precursors such as carotenoids, when exposed to heat, light, in methanol extract of saffron, has been elucidated by Fourier oxygen or the enzyme β-glucosidase. transform-IR and Raman spectroscopy and also crystal structure analysis (Tarantilis et al., 1994c). Bitter compounds

Volatiles The colourless glycoside picrocrocin (C16H26O7, 4-(β-D- glucopyranosyloxy)-2, 6, 6-trimethyl-1-cyclohexene-1- Literature revealed that saffron contains more than 150 volatile, carboxaldehyde) is the major bitter compound of saffron. The non-volatile and aroma-yielding compounds including of 40-50 compound was first isolated by Winterstein and Teleczky in 1922. volatiles components with strong odour (Melnyk et al., 2010). Kuhn and Winterstein (1934) examined its chemical properties. UV Tarantilis and Polissiou reported the presence of 23 volatile absorption maxima for picrocrocin was observed at 254 nm/ε = components, 13 of which were ketones and 6 aldehydes. The 7124 (Alonso et al., 1999a) and 250.5 nm/ε = 10100.The formation molecular weights of these compounds ranged from 122–208 and of picrocrocin is related to degradation of zeaxanthin. Its their presence was directly related to the method of isolation of decomposition gives rise to compounds responsible for the aroma of volatile compounds. Of them namely, 2,6,6-trimethyl-1,3- saffron. Removal of the sugar moiety takes place during processing cyclohexadien-1-carboxaldehyde (safranal) (70%); 3,5,5-trimethyl- (drying, storage) of saffron or even during flower development. The 2-cyclohexen-1-one (isophorone) (14%); 3,5,5,-trimethyl-3- hydrolysis of picrocrocin may also occur due to activation of β- cyclohexene-1-one (isomer of isophorone)(5%); 2,6,6-trimethyl-2- glucosidase. Iborra et al. (1992a) proposed a method for the cyclohexene-1,4-dione (4%); and 2,6,6-trimethyl-1,4- immobilization of this enzyme in order to increase the rate of cyclohexadiene-1-carboxaldehyde (isomer of safranal) (3%) were hydrolysis yielding higher amounts of safranal. the major compounds. Structures of these compounds were elucidated by mass spectroscopy. Himeno and Sano (1987) studied the picrocrocin content in intact stigmas of saffron during the flower development and observed a These volatiles with a common nucleus possibly originate from stepwise increase in the content before anthesis. Then, an picrocrocin and are formed either by enzymatic activity or by appreciable decrease in picrocrocin level coincided with the oxidative degradation process. A scheme for the formation of appearance of volatiles. Alonso et al. (2001) who examined several safranal is presented in Fig.2. samples of Spanish, Indian and Iranian saffron found their picrocrocin content as 0.79–12.94% in Spanish 1.07–2.16% in Indian and 2.18–6.15% in Iranian varities. Syed Muzaffar et al. /Int J Biol Med Res.10(4):6910 - 6919 6915

Constituents from other plant organs of C. sativus

Various flavonoids have been found in the flowers, petals and leaves of many Crocus species (Poldini et al., 1979; Harborne and Williams, 1984). Kaempferol, Astragalin, Helichrysoside, Kaempferol-3-O-β-D-glucopyranosyl-(1,2)-β-D-6-acetylglucopyranoside and kaempferol-3- O-β-D-glucopyranosyl-(1,2)-β-D-glucopyranoside myricetin, petunidin, and delphinidin-3,5-diglycoside have been reported in pollen, petals and tepals of C. sativus. Gao et al. (1998) reported on the isolation and characterisation of some phenolic glycosides and a γ-lactone glucoside from the sprouts of the plant. The compounds were characterized as 2, 4-dihydroxy-6-methoxyacetophenone-2-β-D-glucopyranoside,2,3,4- trihydroxy-6-methoxy acetophenone-3-β-D-glucopyranoside and 3-(S)-3-β-D-glucopyranosylbutanolide. Escribano et al. (1999a) isolated a proteoglucan of 30 kDa from corms of C. sativus. The polysaccharide part of the molecule was found to contain 36.4% rhamnose, while the protein backbone is composed of aspartic acid/asparagine, alanine, glutamic acid/glutamine, glycine and serine, which make up 60% of the total amino acids.

Adulteration of Saffron

Saffron has been adulterated in various ways since beginnings in order to obtain benefits because of its high cost. In fact, in the time of Marco Polo (13th century) its price was higher than gold. Pliny said (1st century), “Nothing is adulterated as much as saffron” (Negbi, 1999). Adulterations have been carried out with a large variety of animals, vegetable and mineral materials (Table 3) 6916 Syed Muzaffar et al. /Int J Biol Med Res.10(4):6910 - 6919

The Dehydration process used as a tonic and a promoter of non-specific immunological defence. In the Chinese therapeutics it is used to treat some During the dehydration process for obtaining saffron spice, disorders of the central nervous system (Schmidt et al., 2007). Crocus sativus L. stigmas lose 80% of their weight. To preserve Various pharmacological activities such as anticonvulsant activity, saffron for a long time its moisture content must be reduced to the anticancer activity, antinociceptive activity, anti-inflammatory 12% as established by the ISO/TS 3632 specification, or even lower effect, anti-depressant, relaxant activity, antihypertensive activity, than that. According to dealers, the organoleptic differences that antioxidant activity, antitussive activity and antihyperlipidemic they detect in saffron from different origins is caused by the activity have been reported (Melnyk et al., 2010). Defence against dehydration process and not by genetic or edaphoclimatic oxygen toxicity, effect in improving ocular blood flow and retinal differences. function, inhibitory effect on Platelet aggregation, radical scavenging activity, antityrosinase activity, apoptosis effect, catalase activity, Simplifying things, it can be said that there are two forms of enzymatic activity, effect on Uterus and Estrus Cycle, glutathione S- saffron dehydration. The first one consists of drying at room transferase (GST) activity, immunomodulating activity, modulatory temperature, by spreading the stigmas out over large surfaces, effects have also been reported (Tavakkol-Afshari et al., 2008; Wang exposing them to the sun or in the shade, in a ventilated place. This et al., 1996). process is used in countries such as India, Iran and Morocco. In India stigmas are dried in the sun for 3-5 days until humidity content is Saffron has a high LD50 = 20 g/Kg which explains why toxicology reduced to 8-10 % (Nauriyal et al., 1977; Sampathu et al., 1984). In researchers currently consider it to be safe for human consumption Kashmir the usual procedure is to dry the whole flower in the sun (Bisset and Wichtl, 1994). The medicinal use of saffron and other and then normally separate the three filaments from the stigma herbs and spices has declined in the 20th century but since 90s (Hassnain, 1998). In Morocco stigmas are placed on a cloth in very saffronologists enhanced their efforts in this direction. Abdullaev thin layers and dried for two hours in the sun and then 7-10 days in and co-workers reviewed the cytotoxic, anticancer and antitumor shade (Ait-Oubahou and El-Otmani, 1999). properties of saffron and its active gredients in, mainly in, in-vitro as well as in vivo studies. Researchers working on the medical The other possibility is to expose saffron to high temperatures by properties of saffron and other parts of the plant e.g., corms are means of hot air flows or placing it on a source of heat. This is the mainly coming from producing countries: Greece, Spain and from method used in Spain, Greece and Italy. In the latter, the process is countries with long tradition in herbal medicine such as Japan carried out by placing fresh stigmas in a sieve situated some 20 cm (Morjani et al., 1990; Bolhassani et al., 2014; Hill, 2004). Though the above oak embers. Halfway through the process, which lasts active constituents of saffron are considered to be crocins, other between 15 to 20 minutes, these are turned over to assure compounds have not been intensively examined in the case of saffron homogenous drying. It is considered done when stigmas contain but are known to possess significant anti-stress/anti-anxiety moisture between 5 to 20 %. At the point they possess some properties such as mangicrocin. They also significantly contribute to elasticity when squeezed between ones fingers (Tammaro, 1999). the therapeutic properties of the spice ((Deo, 2003; Rau, 1969; Ghosal et al., 1989), as well. In Greece the mixture of fresh stigmas and part of the stamens is speed over in a thin layer of 4-5 mm on 40-50 cm trays with the The use of saffron for textile dyeing declined with the advent of bottom covered by silk cloth. These trays, 25 -30 cm apart, are placed synthetic dyes. Recent papers indicate an interest for dyeing natural on vertical supports in rooms meant for dehydration. During the first expensive textiles (silk, wool, and cotton) with it. Tsatsaroni and few hours the temperature is kept at 200C and later raised to 30 - Eleftheriadis (1994) studied the dyeing and fastness properties of 350C. The drying process ends when moisture is reduced to 10- 11 saffron on cotton and wool. 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