MELISSA OFFICINALIS L.) AND IMPROVEMENT OF ITS GASTRO ... · Melissa officinalis L. dried and ground. • Lemon balm ipowder® is a powder composed of an aqueous extraction of two

ENRICHMENT OF ROSMARINIC ACID FROM LEMON BALM (MELISSA OFFICINALIS L.) AND IMPROVEMENT OF ITS GASTRO-INTESTINAL BIOACCESSIBILITY BY USING THE TECHNOLOGY Michel Dubourdeaux 1 , Anaïs Escalon 1 , Valérie Bardot 1 , César Coe 1 , Sylvain Denis 2 , Monique Alric 2 , Sandrine Chalancon 2 , Isabelle Ripoche 3 , Pierre Chalard 3 , Lucile Berthomier 3 , Marn Leremboure 3 1 Groupe Pileje, Paris, France, [email protected]; 2 Université Clermont Auvergne, INRA, UMR454 MEDIS, F-63000 Clermont-Ferrand, France; 3 Université Clermont Au- vergne, CNRS, SIGMA Clermont, Clermont-Ferrand, France MATERIAL & METHODS RESULTS CONCLUSION CONFLICT OF INTEREST DISCLOSURE: V Bardot, A Escalon, C Coe, M Dubourdeaux are employees of PiLeJe Industrie. I Ripoche, P Chalard, L Berthomier, and M Leremboure performed the chromatographic analyses and S Denis, M Alric, S Chalancon performed the expe- riments in the TIM model for PiLeJe. 2 Quantification of rosmarinic acid • ipowder® contains a higher concentraon of RA than the nave plant. 3 Release of rosmarinic acid (RA) during in vitro gastro-intestinal digestion • Compared to RA total amount in each extract, recovery percentage of RA was higher with ipowder® than nave plant ( a ANOVA, p<0.05). • 3 capsules of the nave plant (900 mg in total) were necessary to release the same amount of RA given by only one capsule of ipowder® (300 mg) Nave plant (3 capsules) ipowder® (1 capsule) Total RA ± SD (mg) % of intake ± SD Total RA ± SD (mg) % of intake ± SD Intake 12.44 ± 0.02 100 7.79 ± 0.10 a 100 Cumulated dialysates 7.5 ± 0.7 60.0 ± 5.3 6.6 ± 0.4 85.3 ± 4.8 a Cumulated ileal effluents 0.07 ± 0.02 0.6 ± 0.2 0.18 ± 0.06 a 2.3 ± 0.7 a Final GIT content 0.27 ± 0.04 2.2 ± 0.3 0.25 ± 0.24 3.2 ± 3.1 Total release 7.8 ± 0.6 62.7 ± 5.1 1.0± 0.5 90.8 ± 6.5 a [1] Shakeri A, Sahebkar A, Javadi B. 2016. Melissa officinalis L. A review of its tradional uses, phytochemistry and phar - macology. J Ethnopharmacol. 188:204–28. [2] Michel Dubourdeaux; EP2080436 A2, 2009. [3] Blanquet S, et al. 2004. A dynamic arficial gastrointesnal system for studying the behavior of orally administered drug dosage forms under various physiological condions. Pharm Res. 21(4):585-91. ABSTRACT Raw materials • Nave plant: leaves of Melissa officinalis L. dried and ground. • Lemon balm ipowder® is a powder composed of an aqueous extracon of two parts of plant followed by concentraon, vacuum fixaon and drying of this extract on one part of plant material. TNO gastro-Intesnal Model (TIM-1) The TIM-1 is a mul-compartmental and dynamic system reproducing the human digesve process. 1 capsule of ipowder® or 3 capsules of nave plant were digested with 200 mL of mineral water. Dia- lysis fluids were regularly collected during digeson and aﬅer 4 h, final contents of the four com- partments (stomach, duodenum, jejunum and ileum) were pooled. Volumes were measured and samples stored at -20°C for downstream analysis of RA concentraon. Phytochemical fingerprint by HPTLC: CAMAG equipment RA quanficaon by HPLC: WATERS system 1 Phytochemical fingerprint Image obtained aﬅer derivazaon with Neu PEG at 366 nm. 1 - Rosmarinic acid 2 - Nave plant 3 - ipowder® Advantages Nave plant Lemon balm ipowder® a a a a This exclusive and patented process insures : 1 dose ipowder® 3 doses nave plant Introducon: Lemon balm (Melissa officinalis L.) is a plant that has long been used in tradional medicine for the treatment of numerous diseases[1]. It contains high level of rosmarinic acid (RA) a well-known bioacve polyphenol. Lemon balm ipowder® is a plant infusion concentrated on plant material, an exclusive 100% plant product made by PiLeJe Industrie from dried M. officinalis L. according to a patented process[2]. Study objecve: The objecve was to assess the benefits of the ipowder® technology applied to M. officinalis L. by comparing rosmarinic acid (RA) release from the ipowder® to the nave plant during in vitro gastro intesnal digeson. Methods: Quanficaon of RA in raw materials or digeson samples was analyzed by HPLC. In vitro gastro intesnal digeson was conducted with the TNO gastro-Intesnal Model (TIM-1), a dynamic 4 stage system simulang the human digesve process[3]. One capsule of lemon balm ipowder® (300 mg) or three of the nave plant (ground dried leaves; 900 mg) were digested 3 mes with 200 mL of mineral water in TIM-1. Dialysis fluids from jejunal and ileal contents and ileal effluents were collected during 4h digesons to assess the total amount of RA released. Results: RA concentraon in the ipowder® was twice higher than in the nave plant. In vitro digesons demonstrated that: 1) three capsules of leaves were necessary to release the same amount of RA given by one capsule of ipowder® and 2) the percentage of RA released was significantly increased with the ipowder® (90.8 ± 6.5%) compared to the nave plant (62.7 ± 5.1%; ANOVA, p<0.05). Conclusion: The ipowder® technology concentrates RA from M. officinalis L. and RA was more available for intesnal absorpon with the ipowder® than the nave plant: there was a 3-fold reducon in the quanty of ingested product to obtain the same amount of RA delivered into the upper gastro-intesnal tract. Therefore, the ipowder® technology allows reducing the dosage for such food supplement which is more comfortable for consumers.

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MELISSA OFFICINALIS L.) AND IMPROVEMENT OF ITS GASTRO ... · Melissa officinalis L. dried and ground. • Lemon balm ipowder® is a powder composed of an aqueous extraction of two