Quantitation of Crocins and Picrocrocin in Saffron by HPLC: Application to Quality Control and Phytochemical Differentiation from Other Crocus Taxa

Abstract

A chromatographic method was developed and fully validated for the determination of the major saffron constituents, i.e., picrocrocin and five major crocins. Dried samples (styles of Crocus sativus and other Crocus taxa) were extracted with MeOH : water (1 : 1, v/v), and chromatographic separation of the analytes was achieved by reversed-phase chromatography using a gradient elution. Full validation was performed using spiked samples with analytes, which were isolated, purified, and characterized by MS due to a lack of commercial standards. The method showed a good fit (r2 > 0.999) for all analytes with limit of quantitation values in the range of 1-15 µg/mL, and demonstrated adequate intra- and inter-precision (< 15 % RSD) and accuracy (< 7 % RE). The method was applied to the analysis of various commercial saffron samples and of indigenous Crocus taxa and allowed for the first time the absolute quantitation of several Crocus components.

Rapid isolation and characterization of crocins, picrocrocin, and crocetin from saffron using centrifugal partition chromatography and LC–MS

“…The whole procedure yielded totally 35.4 g of the C. sativus extract (59% of 60 g starting material). This result is in accordance with the extraction yield described in previous experiments [9].…”

“…Over the past years, there have been many studies, regarding the major and minor compounds of C. sativus . The main saffron constituents (Figure ) include crocins, which are mono‐ and bis‐esters of crocetin (apocarotenoid) with glucose and/or gentiobiose, and monoterpenes, mainly picrocrocin and safranal . Picrocrocin, a colorless glycoside, is the β ‐ d ‐glucoside of hydroxysafranal (4‐hydroxy‐2,6,6‐trimethyl‐1‐cyclohexene‐1‐carboxaldehyde) and mainly responsible for saffron's well‐known bitterness .…”

“…Saffron is considered to be the most expensive spice that is mainly used as traditional herbal medicine or food coloring Figure 1) include crocins, which are mono-and bis-esters of crocetin (apocarotenoid) with glucose and/or gentiobiose, and monoterpenes, mainly picrocrocin and safranal. [9] Picrocrocin, a colorless glycoside, is the β-D-glucoside of hydroxysafranal (4-hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde) and mainly responsible for saffron's well-known bitterness [10]. Safranal is the principal substance of saffron's aroma.…”

Efficient production of saffron crocins and picrocrocin inNicotiana benthamianausing a virus-driven system

“…Crocins reported values in saffron samples from Spain ranged from 0.85 to 32.40 mg/g dry weight (ALONSO et al, 2001). Other reports showed 24.87 mg/g from China (Tong et al ., 2015), 26.60 mg/g from Greece (Koulakiotis et al, 2015), 29.00 mg/g from Morocco (Gonda et al, 2012), 32.60 mg/g from Iran, 49.80 mg/g from Italy (Masi et al, 2016) or 89.00 mg/g from Nepal (Li et al, 2018). In Gardenia fruits, crocins levels range from 2.60 to 8.37 mg/g (Ouyang et al, 2011; Wu et al, 2014).…”

“…The picrocrocin content ranges from 7.90 to 129.40 mg/g in Spanish saffron. In Greek saffron, 6.70 mg/g were reported (Koulakiotis et al, 2015), and 10.70-2.16 mg/g in Indian, 21.80-6.15 mg/g in Iranian and 42.20-280.00 mg/g in Moroccan saffron (Lage and Cantrell, 2009). The different values obtained in those samples are also a consequence of the different procedures followed to obtain the saffron spice.…”