Development of methods for standardization of Crocus sativus (saﬀron) stigmas for inclusion in the draft monograph of the State Pharmacopoeia of Ukraine
Authors: Mykhailenko O. O. , Kotov A. G. , Kotova E. E. , Sira L. M. , Kovalyov V. M. , Georgiants V. A. 
Published: April 2021

Crocus sativus L. from the Iridaceae family is a medicinal and edible plant that has recently been actively cultivated in Ukraine. Saﬀron spice is crocus ﬂower stigmas exhibiting a wide range of the pharmacological activity due to its three main bioactive compounds: crocin, picrocrocin and safranal. The quality of this raw material is regulatedby various normative documents, but there is no monograph in the State Pharmacopoeia of Ukraine (SPhU).
Aim. To perform a comparative analysis of the monograph “Saﬀron for homoeopatic preparations” from the Ph. Eur. 9.0, “Saﬀron. Croci stigma” from the Deutscher arzneimittel codex (DAC) and “Spices – Saﬀron (Crocus sativus L.)” from the ISO 3632 by the following indicators: description, identifcation (microscopy and TLC), impurities, loss on drying, total ash and quantifcation of crocin, picrocrocin and safranal by UV-Vis-spectrophotometry to clarify the possibility of harmonizing the requirements of the national legal framework for Crocus stigmas withthe Ph. Eur. 9.0, DAC and ISO 3632. The results of the analysis will be taken into account in the development of the monograph of the national part of the SPhU “Crocus stigmas (saﬀron)”.Results and discussion. The quality indicators of Crocus stigmas and their standardization determinedby the monograph in the Ph. Eur. 9.0, DAC and ISO 3632 on the raw material studied have been analyzed; some diﬀerences in the regulated quality indicators of the raw material have been found. The results of our own research have shown that the samples of the Ukrainian Crocus (saﬀron) meet all these requirements. The results of the macro- and microscopic examination of the raw material are given. The chromatographic examination hasbeen performed in accordance with the Ph. Eur. 9.0 and DAC using crocin as a standard compound; the quantitative determination of crocin, picrocrocin and safranal in 8 samples of Crocus stigmas has been performed by UV-Vis-spectrophotometry in accordance with the ISO 3632. It has been proposed to introduce the following indicators to determine the specifc absorption rate: for crocin (C44H64O24; Mw – 976.70) not less than 180, for safranal (C10H14O; Mw – 150.22) – not less than 30, for picrocrocin (C16H26O7; Mw – 330.37) – not less than 60 calculated with reference to the dried raw material.Experimental part. For analysis the dried stigmas of Crocus sativus (saﬀron) collected in the villageLyubimivka, Kherson region, Ukraine (2016-2018), as well as commercial samples of saﬀron from Morocco,Azerbaijan, Spain were used. The analysis of the leading normative documents containing monographs “Crocus stigmas”, among them the Eur. Ph. 9.0, DAC and ISO 3632, was performed. The studies included description of the appearance of the whole Crocus stigmas; loss on drying and the mass fraction of volatile compounds (105 °C, 16 h); the mass fraction of total ash. The amount of picrocrocin, safranal and crocin was measured by direct reading of the absorption of 1 % aqueous solution of stigmas at 257 nm, 330 nm and 440 nm, respectively, usinga UV-Vis-spectrophotometer.Conclusions. The analysis of the requirements to quality of the medicinal raw material – stigmas of Crocus sativus – has been performed based on the monographs: “Saﬀron for homoeopatic preparations” from the Ph. Eur. 9.0, “Saﬀron. Croci stigma” from the Deutscher arzneimittel codex and “Spices – Saﬀron (Crocus sativus L.)” from the ISO 3632. The main criteria for standardization of the raw material have been determined. It has been proposed to introduce the monograph to the national part of the SPhU “Crocus stigmas (saﬀron)” according to the following requirements: identifcation (macro- and microscopic signs; thin-layer chromatography (crocin); the quantitativedetermination (the content of crocin, picrocrocin and safranal) adapted to the ISO 3632, performed by UV-Vis-spectrophotometry; impurities; loss on drying; total ash.Key words: standardization; State Pharmacopoeia of Ukraine; Crocus sativus stigma; saﬀron; crocin