Elucidation of metabolite isomers of Leonurus japonicus and Leonurus cardiaca using discriminating metabolite isomerism strategy based on ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry.

The isomer structural discrimination is a significant challenge in metabolome analysis based on ultrahigh performance liquid chromatography tandem high-resolution mass spectrometry (UHPLC-HRMS). In this study, a new discriminating metabolite isomerism strategy is proposed to elucidate the metabolome, especially the isomers, of Leonurus japonicus and Leonurus cardiaca. This strategy consists of three steps. First, the metabolite biosynthesis pathways are constructed based on a home-built compound database to rapidly profile the compounds of interest using the multiple diagnostic product ions (DPIs) screening analysis and binary comparison based on SUMPRODUCT function. Second, the fragmentation patterns (e.g. the high-resolution DPIs, DPI ratios) and chromatographic elution order are defined based on scattered reference chromatographic and mass spectrometry data, calculated lipophilicity parameters, molecular hydrogen bond analysis, and chemical reference standards. Finally, all discovered isomerisms are mapped with the defined applicable rules and the isomers are identified conveniently. Using this strategy, a total of 257 compounds were tentatively characterized, including 212 potential novel compounds and 67 pairs of cis-, trans-, and positional isomers of flavonoids, phenylethanoid glycosides, glucaric acids, novel quinic acids, and esters of fatty acids. Moreover, 56 characteristic markers were identified to discriminate these two herbal medicines. This strategy may significantly improve the efficiency and reliability of identifying isomers found in metabolite biosynthesis pathways.

REPRESENTATIVES OF MOTHERWORT GENUS (LEONURUS SPP.): ASPECTS OF PHARMACOGNOSTIC FEATURES AND RELEVANCE OF NEW SPECIES APPLICATION.

The review deals with analysis and classification of literature data on phamacognostic study (botanical characteristics, geographical distribution, chemical composition and use in medicine) of species of Motherwort genus (Leonums L.). The review unveils opportunities for comprehensive study and development of approaches to the standardization of raw materials of Turkistan motherwort (L. turkestanicus V.I. Krecz. & Kuprian). That will allow to expand the range of sources of herbal remedies raw materials in the framework of realization of the state program for import substitution.

GC-MS and q-NMR based chemotaxonomic evaluation of two Leonurus species.

INTRODUCTION: The genus Leonurus L. (fam: Lamiaceae) is represented in Uzbekistan by two species, L. panzerioides Popov. and L. turkestanicus V. I. Krecz. & Kuprian, which are used to treat nervous disorders and also as sedative and hypotensive agents. OBJECTIVES: To establish the taxonomic status of Leonurus panzerioides and L. turkestanicus based on their chemical constituents analysed by GC-MS and q-NMR. MATERIALS AND METHODS: Quantitative (1) H-NMR (q-NMR) was used to identify and quantify known major components in the methanol extracts of these two species. Additionally, the chemical composition of the essential oils obtained from the aerial parts of these plants were analysed by GC-MS. RESULTS: The q-NMR analyses of Leonurus panzerioides and L. turkestanicus revealed the presence of 8-acetylharpagide, harpagide, leonurine and stachydrine as major components. Using the GC-MS method, overall 24 and 39 constituents were identified, respectively, from L. panzerioides and L. turkestanicus oils. The major constituents of the essential oil of L. panzerioides were eugenol (30.9%) and p-vinyl guaiacol (15.8%), whereas thymol (40.1%) and octen-3-ol (13.1%) were the principal compounds in the essential oil of L. turkestanicus. CONCLUSION: The major components in Leonurus panzerioides and L. turkestanicus as identified by the GC-MS and q-NMR analyses, were similar to those present in other Leonurus species and thus provided chemotaxonomic evidence for the placement of these species under the genus Leonurus. Copyright © 2016 John Wiley & Sons, Ltd.

[Genetic relationship analysis of different provenances of Leonurus japonicus by ISSR marker].

OBJECTIVE: To study the genetic polymorphism and genetic relationship of different provenances of Leonurus japonicus. METHOD: Genetic relationship in 19 provenances, which came from nationwide origins were measured by using polymerase chain reaction (PCR) with ISSR marker. RESULT: Twenty ISSR primers were selected from 100 ISSR primers and used for ISSR amplification. A total of 164 bands were generated, of which 117 bands were polymorphic bands (the percentage of polymorphic band, PPB = 71.34%). The similarity coefficients were calculated with NTsys 2.10e software and the dendrogram were constructed with UPGMA. Nineteen samples were divided into 3 types using the cluster analysis according to their genetic similarity coefficient. CONCLUSION: Results from the cluster analysis were correlated significantly with the morphological characteristics and geographical location of 19 samples. The data indicate that ISSR technique is useful to determine genetic diversity and genetic relationship among Leonurus provenances, providing a scientific basis for genetic breeding, differentiation and new cultivar selection.

[Nuclear ribosomal DNA internal transcribed spacer 1 sequences of 4 Leonurus species].

OBJECTIVE: To analyze the nuclear ribosome DNA (nrDNA) internal transcribed spacer (ITS) sequences of 4 Leonurus species, and the possibility of using them for molecular authentication of the crude drugs from the genus. METHODS: The nrDNA ITS sequence (including ITS1, 5.8S rDNA, ITS2, and partial 18S rDNA and 26S rDNA) of L. japonicus and its 3 adulterant species were amplified and sequenced, and CLUSTRAL X and MEGA software was employed for analysis. RESULTS: The variation of ITS1 and ITS2 between L. japonicus and its adulterant species ranged between 7.2% and 18.8% and between 14.2% and 27%, respectively. The phylogenic tree derived from the dendrograms based on the ITS sequence data contained some discrepancy from the traditional classification. CONCLUSION: The nrDNA ITS sequences can be used potentially as efficient markers for identification of L. japonicus and its adulterants, and further study is needed for studying the phylogeny of Leonurus.

[The descriptions identification of "yimu cao"].

Remained but entire bracts beneath the inflorescence and the persistent calyxes were selected as primary characteristics to distinguish "Yimu Cao", which derived from 8 species and 1 variety in the same genus Leonurus. The results were reliable.

[Identification on crude drugs from genus Leonurus: II. Leaf and flower surface characters].

OBJECTIVE: To identify the crude drugs originated from 9 species and 1 variety of genus Leonurus (called Yimucao) through leaf and flower surface characters. METHOD: Routine slide processing and the characters under microscope were observed and compared. RESULTS: Obvious characteristics for identification were revealed. CONCLUSION: The shape and length of non-glandular hairs are useful in identifying this sort of crude drugs. Leaf constants and cluster crystals are also useful as reference. However, pollen grains and secretory tissues are worthless.

[Identification of crude drugs from genus Leonurus].

The structure in cross section of stem and lamina of 9 species and 1 variety from genus Leonurus, whose herb used as Yimu Cao, were reported. Diagnostic characteristics were listed out in a key for identification.