Qualitative and Quantitative Analysis of Secondary Metabolites in Morphological Parts of Paulownia Clon In Vitro 112® and Their Anticoagulant Properties in Whole Human Blood

Abstract

It is not easy to find data in the scientific literature on the quantitative content of individual phytochemicals. It is possible to find groups of compounds and even individual compounds rather easily, but it is not known what their concentration is in cultivated or wild plants. Therefore, the subject of this study was to determine the content of individual compounds in the new Paulownia species, Oxytree, developed in a biotechnology laboratory in 2008 at La Mancha University in Spain. Six secondary metabolites were isolated, and their chemical structure was confirmed by spectral methods. An analytical method was developed, which was then used to determine the content of individual compounds in leaves, twigs, flowers and fruits of Paulownia Clon in Vitro 112®. No flavonoids were found in twigs and fruits of Oxytree, while the highest phenylethanoid glycosides were found in twigs. In this study, we also focused on biological properties (anticoagulant or procoagulant) of extract and four fractions (A–D) of different chemical composition from Paulownia Clon in Vitro 112 leaves using whole human blood. These properties were determined based on the thrombus-formation analysis system (T-TAS), which imitates in vivo conditions to assess whole blood thrombogenecity. We observed that three fractions (A, C and D) from leaves decrease AUC10 measured by T-TAS. In addition, fraction D rich in triterpenoids showed the strongest anticoagulant activity. However, in order to clarify the exact mechanism of action of the active substances present in this plant, studies closer to physiological conditions, i.e., in vivo studies, should be performed, which will also allow to determine the effects of their long-term effects.

Paulownia Organs as Interesting New Sources of Bioactive Compounds

“…Currently, there are few publications describing the chemical constituents of Paulownia Clone in vitro 112. As the chemical content varies between different species and cultivars, more research is needed to ascertain the content of secondary metabolites and their properties [ 4 , 113 ].…”

“…Overall, the extract had stronger antiplatelet activity than the fractions [ 24 ]. Moreover, fraction D showed the strongest anticoagulant activity in whole blood, which was determined with the Total Thrombus-Formation Analysis System (T-TAS) [ 113 ].…”

Identification and Pharmacological Activities of Secondary Metabolites of Phytochemicals

“…Both techniques have been used to separate, identify and quantify series of structurally different SMs in plants include spectrophotometric [57,[73][74][75], GC-MS [56,76], LC-MS [77], LC-MS/MS [56,76,78], Liquid Chromatography-High Resolution Electrospray Ionization Mass Spectrometry (LC-HRESIMS) [57], Nuclear Magnetic Resonance (NMR) [44,79], Ultra-Fast Liquid Chromatography (UFLC-Q-TOF-MS/MS) [80][81], UFLC-triple-Q-TOF-MS/MS [82], Ultra-high performance Liquid Chromatography quadrupole-time of flight mass spectrometry [83], UHPLC-LTQ-Orbitrap [84], UHPLC/Q-Orbitrap-MS [85], HPLC-DAD [86], UHPLC-Q-TOF-MS [87], UHPLC-Q-TOF-MS/MS [88], HPLC-DAD-ESI-MS [89], UPLC-QTOF/MSE [90], UHPLC-MS/MS [91], HPLC-ESI-QTOF-MS [92], UPLC-QTOF-MS/MS [93], UHPLC-Q-TOF-MS/MS [94][95], UHPLC-ESI-MS/MS [96], UHPLC-ESI-Orbitrap-MS/MS [97], UPLC-IT-MS [98], UPLC-Q-TOF-MS [98], UPLC-PDA [45], HPLC-ESI-MS and MS 2 [99], reverse phase high performance liquid chromatography-diode array detector (RP-HPLC-DAD) [74], HPLC-HRMS [100]. been used to produce a yellow color that turn colorless on addition of dilute hydrochloric acid confirm flavonoid [102,[104][105]…”

Effect of Paulownia Leaves Extract Levels on In Vitro Ruminal Fermentation, Microbial Population, Methane Production, and Fatty Acid Biohydrogenation

“…Recently, many biologically active substances and tree leaves have been used as a source of nutrients and plant bioactive compounds to decrease methane production and improve ruminal fermentation [3,8,9]. An example could be a Paulownia tree [10]. Paulownia dried leaves or silage positively affected the rumen fermentation parameters by changing acetate to a propionate ratio without any adverse effect on nutrient digestion, which could be exploited in the nutrition of ruminants [3,9].…”