Enhanced accumulation of harpagide and 8-O-acetyl-harpagide in Melittis melissophyllum L. agitated shoot cultures analyzed by UPLC-MS/MS

Abstract

Harpagide and its derivatives have valuable medicinal properties, such as anti-inflammatory, analgesic and potential antirheumatic effects. There is the demand for searching plant species containing these iridoids or developing biotechnological methods to obtain the compounds. The present study investigated the effects of methyl jasmonate (MeJa, 50 μM), ethephon (Eth, 50 μM) and L-phenylalanine (L-Phe, 2.4 g/L of medium), added to previously selected variant of Murashige and Skoog medium (supplemented with plant growth regulators: 6-benzylaminopurine 1.0 mg/L, α-naphthaleneacetic acid 0.5 mg/L, gibberellic acid 0.25 mg/L) on the accumulation of harpagide and 8-O-acetyl-harpagide in Melittis melissophyllum L. agitated shoot cultures. Plant material was harvested 2 and 8 days after the supplementation. Iridoids were quantitatively analyzed by the UPLC-MS/MS method in extracts from the biomass and the culture medium. It was found that all of the variants caused an increase in the accumulation of harpagide. In the biomass harvested after 2 days, the highest harpagide content of 247.3 mg/100 g DW was found for variant F (L-Phe and Eth), and the highest 8-O-acetyl-harpagide content of 138 mg/100 g DW for variant E (L-Phe and MeJa). After 8 days, in some variants, a portion of the metabolites was released into the culture medium. Considering the total amount of the compounds (in the biomass and medium), the highest accumulation of harpagide, amounting to 619 mg/100 g DW, was found in variant F, and the highest amount of 8-O-acetyl-harpagide, of 255.4 mg/100 g DW, was found in variant H (L-Phe, MeJa, Eth) when harvested on the 8th day. These amounts were, respectively, 24.7 and 4.8 times higher than in the control culture, and were, respectively, 15 and 6.7 times higher than in the leaves of the soil-grown plant. The total amount of the two iridoids was highest for variant F (0.78% DW) and variant H (0.68% DW) when harvested on the 8th day. The results indicate that the agitated shoot cultures of M. melissophyllum can be a rich source of harpagide and 8-O-acetyl-harpagide, having a potential practical application. To the best of our knowledge we present for the first time the results of the quantitative UPLC-MS/MS analysis of harpagide and 8-O-acetyl-harpagide in M. melissophyllum shoot cultures and the enhancement of their accumulation by means of medium supplementation with elicitors and precursor.

Matrix Fmea Analysis as a Preventive Method for Quality Design of Hydraulic Components

“…Such approach often requires special prediction models (Pietraszek et al, 2014;Pietraszek et al, 2017a) with an uncertainty estimated by non-parametric methods (Pietraszek and Gadek-Moszczak, 2013;Gadek-Moszczak et al, 2015;Dwornicka et al, 2017;Pietraszek et al, 2017b). Strong support for this approach is offered by a design of experiments methodology (Montgomery, 2008) which is widely used from artificial neural networks (Pietraszek, 2003) through a biotechnology (Skrzypczak-Pietraszek et al, 2018a), a phytochemistry (Skrzypczak- Pietraszek and Pietraszek, 2009;Skrzypczak-Pietraszek et al, 2018b), a mechanics (Kozien, 2009;Goroshko et al 2014) up to a industry management (Ulewicz et al, 2013;Ulewicz, 2016;Kozien and Kozien, 2017;Maszke et al, 2018). Pietraszek, J., Jasiewicz, B., Sikorska, S., Wojnar, L., 2015. The bootstrap approach to the comparison of two methods applied to the evaluation of the growth index in the analysis of the digital X-ray image of a bone regenerate.…”

Hydraulic Systems Safety by Reducing Operation and Maintenance Mistakes

“…To achieve maximum effectiveness, it is recommended to use a design of experiment methods like e.g. in biotechnology (Skrzypczak et al, 1994), phytochemistry (Skrzypczak-Pietraszek and Pietraszek, 2009;Skrzypczak-Pietraszek et al, 2018a;Skrzypczak-Pietraszek et al, 2018b) or materials science (Kuscer et al, 2007;Korzekwa et al,. 2016).…”

Theimpact of the Coatingsystem for Rolling Stock on Fire and Anti-Corrosion Properties

“…In the next step, it is planned to perform more resistance tests: humidity resistance, combine corrosion tests and start long-term natural weathering test. It will include various method such as an image analysis (Gadek-Moszczak, 2017), a fuzzy assessment of an uncertainty (Pietraszek, 2012;Pietraszek et al, 2018), a non-parametric modelling of processes (Pietraszek and Gadek-Moszczak, 2013;Gadek-Moszczak et al, 2015) and, at last, the combination of all these methods inside the design of experiment methodology (Skrzypczak- Pietraszek and Pietraszek, 2009;Skrzypczak-Pietraszek et al, 2018a, Skrzypczak-Pietraszek et al, 2018b. It also seems that a FEM-based numerical simulation of the flow (Domagala et al, 2018a;Domagala et al, 2018b) and thermomechanical processes accompanying the fire burning should provide a lot of detailed information, especially related to properties of construction materials and their mutual interactions (Guidoni et al, 2005;Ulewicz et al, 2010;Hebda et al, 2012;Pieczonka et al, 2012;Kapustka et al, 2017;Gawdzinska et al, 2018).…”