Abstract

In this study, the aphrodisiac activities of Crocus sativus stigma aqueous extract and its constituents, safranal and crocin, were evaluated in male rats.

The aqueous extract (80, 160 and 320mg/kg body wt.), crocin (100, 200 and 400mg/kg body wt.), safranal (0.1, 0.2 and 0.4ml/kg), sildenafil (60mg/kg body wt., as a positive control) and saline were administered intraperitoneally to male rats.

Mounting frequency (MF), intromission frequency (IF), erection frequency (EF), mount latency (ML), intromission latency (IL) and ejaculation latency (EL) were the factors evaluated during the sexual behavior study.

Crocin, at all doses, and the extract, especially at doses 160 and 320mg/kg body wt., increased MF, IF and EF behaviors and reduced EL, IL and ML parameters.

Safranal did not show aphrodisiac effects.

The present study reveals an aphrodisiac activity of saffron aqueous extract and its constituent crocin.

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The Effect of Saffron, Crocus Sativus Stigma, Extract and Its Constituents, Safranal and Crocin on Sexual Behaviors in Normal Male Rats

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Abstract

The stigmas of Crocus sativus L. have been used as spice and colorant agent (i.e. saffron) for more than 4000 years.

For an updated structural investigation of the aglycon present in the glycosylated crocetin apocarotenoids (i.e. crocins), seven representative derivatives ranging from one up to five glucosyl-residues with a maximum number of three monosaccharides per glycosylation site (glucose, gentiobiose, gentiotriose and neapolitanose) were isolated and purified by high-performance liquid chromatography.

The compounds selected for further mass spectrometric investigation include glucosyl-, bis-glucosyl-, gentiobiosyl-, gentiobiosyl-glucosyl-, bis-gentiobiosyl-, gentiobiosyl-gentiotriosyl- and gentiobiosyl-neapolitanosyl-crocetin.

Electrospray ionization in combination with low-energy collision-induced dissociation/tandem mass spectrometry of sodiated crocin precursor ions utilizing either a 3D-ion trap (MS(n) , n = 2, 3) or a QqTOF instrument, with the latter providing accurate mass determination with an accuracy of ±1-3 ppm or better at a resolution of 10,000 (full width at half maximum), was used.

Major fragmentation pathways included loss of either one or two carbohydrate substituents leading to the sodiated aglycon without interglycosidic bond cleavage during in MS(2) -experiments.

All sodiated precursor ions and major product ions were accompanied by a loss of 92 Da, which was elucidated as C7 H8 -loss from the aglycon by skeletal rearrangement via an eight-membered transition state as previously described for intact C40-carotenoids.

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In-chain Neutral Hydrocarbon Loss From Crocin Apocarotenoid Ester Glycosides and the Crocetin Aglycon (Crocus Sativus L.) by ESI-MS(n) (n=2, 3)

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Abstract

Rationale:

The expensive spice saffron originating from the stigmas of Crocus sativus L. and also applied in traditional Chinese medicine (TCM) constitutes a complex mixture of glycoconjugates varying not only in the aglycon structure, but also in glycosylation pattern. Therefore, various tandem mass spectrometric techniques were evaluated for their usefulness in structural elucidation.

Methods:

Three selected constituents of the stigmas of Crocus sativus L., trans- and cis-crocin-4 as well as picrocrocin, were isolated and purified by HPLC and finally analyzed by ESI-MS (ion trap, QqRTOF), IP-MALDI-MS (QqRTOF) and vMALDI-MS (TOF/RTOF) in combination with tandem mass spectrometry in collision energy regimes ranging from a few eV (LE) to 20 keV (HE) collisions for the first time. These data aid in structurally elucidating minor, unknown glycoconjugates originating from this plant-derived spice.

Results:

LE-CID of isomeric crocins on either an ion trap with ESI or a QqRTOF-instrument with ESI or IP-MALDI as desorption/ionization technique only yielded a limited number of structurally diagnostic sodiated product ions related to the carbohydrate moiety as well as to the intact aglycon in contrast to true HE-CID. The low MW constituent picrocrocin did not yield useful LE-CID spectra, but showed a high number of structurally diagnostic product ions by HE-CID utilizing a vMALDI TOF/RTOF-instrument.

Conclusions:

The highest number of structurally diagnostic product ions allowing also determination of the carbohydrate linkage of the gentiobiose-moiety of isomeric crocins ((0,4)A(2), (3,5)A(2) product ions indicating a 1→6 carbohydrate linkage) was only achievable by HE-CID. Fragmentation of the aglycon was not observed by any collision energy regime applied.

Comparison of Different Tandem Mass Spectrometric Techniques (ESI-IT, ESI- And IP-MALDI-QRTOF and vMALDI-TOF/RTOF) for the Analysis of Crocins and Picrocrocin From the Stigmas of Crocus Sativus L

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Abstract

Crocus sativus L. is a plant cultivated in various parts of the world. Crocins are among the active components of Crocus sativus L.

The present study was designed to investigate in the rat whether or not crocins possess anxiolytic properties.

For this aim, the light/dark test was selected.

Either crocins, at a dose which did not influence animals’ motor activity (50mg/kg), or diazepam (1.5 mg/kg), significantly increased the latency to enter the dark compartment and prolonged the time spent in the lit chamber in the rats.

Conversely, lower doses of crocins (15-30 mg/kg) did not substantially modify animals’ behaviour.

The present results indicate that treatment with these active constituents of Crocus sativus L. induce anxiolytic-like effects in the rat.

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Effects of the active constituents of Crocus sativus L., crocins, in an animal model of anxiety

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Abstract

Crocus sativus stigmas are one of the widely known spices (saffron) and consist of unusually polar carotenoids.

Alzheimer’s disease is characterized pathologically by deposition of amyloid beta-peptide (Abeta) fibrils. Oxidation is thought to promote Abeta fibril formation and deposition.

To identify agents inhibiting the pathogenesis of Alzheimer’s disease, we examined in vitro the antioxidant properties of extract of C. sativus stigmas and its effect on Abeta(1-40) fibrillogenesis.

The antioxidant properties were determined by measuring the ferric-reducing antioxidant power and Trolox-equivalent antioxidant capacity, while its effects on Abeta-aggregation and fibrillogenesis were studied by thioflavine T-based fluorescence assay and by DNA binding shift assay.

The water:methanol (50:50, v/v) extract of C. sativus stigmas possesses good antioxidant properties, higher than those of tomatoes and carrots, and inhibited Abeta fibrillogenesis in a concentration and time-dependent manner.

The main carotenoid constituent, trans-crocin-4, the digentibiosyl ester of crocetin, inhibited Abeta fibrillogenesis at lower concentrations than dimethylcrocetin, revealing that the action of the carotenoid is enhanced by the presence of the sugars.

Our findings suggest the possible use of C. sativus stigma constituents for inhibition of aggregation and deposition of Abeta in the human brain.

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Inhibitory Activity on Amyloid-?? Aggregation and Antioxidant Properties of Crocus sativus Stigmas Extract and Its Crocin Constituents

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Radical scavenging activity is involved in aging processes, antiinflammatory, anticancer and wound healing activity.

Hence, in the present study the DPPH radical scavenging activity of a natural product that possesses biological properties, an extract of Crocus sativus L. (saffron), grown in Crocos, Kozani (Greece), and some of its bioactive constituents (crocin, safranal) was studied.

It was shown that a methanol extract of Crocus sativus exhibited high antioxidant activity, although it contains several active and inactive constituents.

In trying to approximate a structure-activity relationship, two bioactive constituents of saffron extract were tested, namely crocin and safranal.

Crocin showed high radical scavenging activity (50% and 65% for 500 and 1000 ppm solution in methanol, respectively), followed by safranal (34% for 500 ppm solution).

All the tested samples showed high radical scavenging activity, probably due to the ability to donate a hydrogen atom to the DPPH radical.

Thus, saffron grown in Greece can be used promisingly in functional foods, drinks with antioxidant activity, in pharmaceutical and cosmetic preparations for their antioxidant activity and probably for their antiaging activity.

Saffron can also be used internally in the form of powder or other pharmacotechnical formulae as a food supplement with antioxidant properties.

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Radical scavenging activity of Crocus sativus L. Extract and its bioactive constituents

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Abstract

The effects of carotenoids of Crocus sativus L. (saffron) on cell proliferation and differentiation of HL-60 cells have been studied and compared with those of all-trans retinoic acid.

Our results demonstrated that the doses inducing 50% inhibition of cell growth were 0.12 microM for all-trans retinoic acid (ATRA) and for carotenoids of saffron 0.8 microM for dimethylcrocetin (DMCRT), 2 microM for crocetin CRT and 2 microM for crocins (CRCs).

At 5 microM, all these compounds induced differentiation of HL-60 cells, at 85% for ATRA, 70% for DMCRT, 50% for CRT and 48% for CRCs.

In these experiments, leukemic cells were cultured for 5 days in the absence or in the presence of up to 5 microM ATRA or seminatural and natural carotenoids.

Since retinoids have a potential application as chemopreventive agents in humans, their toxicity as an important limiting factor for their use in treatment should be extensively explored.

The seminatural (DMCRT and CRT) and natural carotenoids (CRCs) of Crocus sativus L. are not provitamin A precursors and could therefore be less toxic than retinoids, even at high doses.

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Inhibition of Growth and Induction of Differentiation of Promyelocytic Leukemia (HL-60) by Carotenoids From Crocus Sativus L

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