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Influence of isolation techniques on the composition of glucosinolate breakdown products, their antiproliferative activity and gastrointestinal stability of allyl isothiocyanate

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Abstract

Mustard seeds are used as a food, and spice due to their nutritive value, the presence of biologically active compounds, and specific taste. Its morphology was investigated by optical microscopy and scanning electron microscopy before and after isolation by hydrodistillation and microwave assisted distillation. The chemical composition of volatile compounds in different mustard seeds (Brassica juncea L. and Brassica nigra L.) was determined using GC–MS, after applying various techniques (Clevenger hydrodistillation, microwave assisted distillation—MAD, and microwave hydrodiffusion and gravity—MHG). Allyl isothiocyanate, degradation product of allyl glucosinolate, was the main volatile compound in B. juncea essential oils and extract after all applied extraction techniques (91.07–99.01%). Gastrointestinal stability of allyl isothiocyanate was determined employing two-phase digestion model (gastric and duodenal) by two methods (in vitro digestion method using commercial and ex vivo digestion method using human digestive enzymes). The stability rate of allyl isothiocyanate was higher after the gastric digestion phase by both methods. In B. nigra seeds but-3-enyl and allyl isothiocyanate were the main compounds after Clevenger hydrodistillation (80.58% and 15.39%, respectively). After MAD, and MHG the main compounds were 4,5-epithiopentanenitrile (50.70% and 59.93%, respectively), and 3,4-epithiobutanenitrile (7.61% and 25.97%, respectively), originating from the same glucosinolates, i.e. but-3-enyl and allyl glucosinolate, respectively. Antiproliferative activity of mustard seed EOs and extracts was evaluated against human cancer cell lines (MDA-MB-231 and TCCSUP). The best antiproliferative activity was shown for B. nigra MHG extract against MDA-MB-231 cell line with IC50 value of 9.1 µg/mL.

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Abbreviations

EO:

Essential oil

ESP:

Epithiospecifier protein

GSL:

Glucosinolate

GC–MS:

Gas chromatography–mass spectrometry

ITC:

Isothiocyanate

MAD:

Microwave-assisted distillation

MHG:

Microwave hydrodiffusion and gravity

RGE 15:

Rabbit gastric extract 15

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Acknowledgements

We are also thankful for the scientific-research equipment funded by EU grant “Functional integration of the University of Split, PMF-ST, PF-ST and KTF-ST through the development of the scientific and research infrastructure” (KK.01.1.1.02.0018).

Funding

This work has been fully supported by the Croatian Science Foundation within the project "Plants as a source of bioactive sulfur compounds and their ability to hyperaccumulate metals", HRZZ-IP-2016–06-1316.

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Authors and Affiliations

  1. Department of Food Technology and Biotechnology, Faculty of Chemistry and Technology, University of Split, Ruđera Boškovića 35, 21 000, Split, Croatia

    Ivana Vrca & Tea Bilušić

  2. Department of Physics, Faculty of Science, University of Split, Ruđera Boškovića 33, 21000, Split, Croatia

    Josipa Šćurla, Nives Kević & Ivana Bočina

  3. Department of Analytical Chemistry, Faculty of Chemistry and Technology, University of Split, Ruđera Boškovića 35, 21 000, Split, Croatia

    Franko Burčul & Ivica Blažević

  4. Department of Medical Chemistry and Biochemistry, School of Medicine, Šoltanska 2, 21 000, Split, Croatia

    Vedrana Čikeš Čulić

  5. Division of Gastroenterology and Hepatology, University Hospital Split, Spinčićeva 1, 21000, Split, Croatia

    Andre Bratanić

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  1. Ivana Vrca
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, formal analysis, investigations as well as reviewing and editing was performed by all authors. Optical and SEM microscopy: [IV], [JŠ], [NK] and [IB]; isolation of volatile compounds: [IV]; chemistry investigation: [IV] and [IB]; antiproliferative activity (MTT assay): [IV] and [VČČ]; obtaining human gastric juices: [AB]; two-phase digestion simulations: [IV] and [TB]; tandem mass spectrometry analysis: [FB]. The first original draft of the manuscript was written by [IV] and [TB]. All authors have read and approved the final manuscript.

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Correspondence to Ivana Vrca.

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The authors declare that they have no conflict of interest.

Compliance with ethics requirements

The study included an ex vivo method of a digestive model with human digestive juices. Permission for the collection of human digestive juices was obtained by the Ethics Committee of the Clinical Hospital Center—Split and the Ethics Committee of the Faculty of Medicine, University of Zagreb, Reg. No.: 380-59-10106-20-111/105, Class: 641-01/20-02/01.

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Vrca, I., Šćurla, J., Kević, N. et al. Influence of isolation techniques on the composition of glucosinolate breakdown products, their antiproliferative activity and gastrointestinal stability of allyl isothiocyanate. Eur Food Res Technol 248, 567–576 (2022). https://doi.org/10.1007/s00217-021-03903-x

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