The Analysis of Leontopodium leontopodioides (Willd.) Beauv. Chemical Composition by GC/MS and UPLC-Q-Orbitrap MS

Fu S., Cheng R., Deng Z., Liu T.

The Lianhua Qingwen (LHQW) capsule is a popular traditional Chinese medicine for the treatment of viral respiratory diseases. In particular, it has been recently prescribed to treat infections caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, due to its complex composition, little attention has been directed toward the analysis of chemical constituents present in the LHQW capsule. This study presents a reliable and comprehensive approach to characterizing the chemical constituents present in LHQW by high-performance liquid chromatography-Q Exactive-Orbitrap mass spectrometry (HPLC-Q Exactive-Orbitrap-MS) coupled with gas chromatography-mass spectrometry (GC-MS). An automated library alignment method with a high mass accuracy (within 5 ppm) was used for the rapid identification of compounds. A total of 104 compounds, consisting of alkaloids, flavonoids, phenols, phenolic acids, phenylpropanoids, quinones, terpenoids, and other phytochemicals, were successfully characterized. In addition, the fragmentation pathways and characteristic fragments of some representative compounds were elucidated. GC-MS analysis was conducted to characterize the volatile compounds present in LHQW. In total, 17 compounds were putatively characterized by comparing the acquired data with that from the NIST library. The major constituent was menthol, and all the other compounds were terpenoids. This is the first comprehensive report on the identification of the major chemical constituents present in the LHQW capsule by HPLC-Q Exactive-Orbitrap-MS, coupled with GC-MS, and the results of this study can be used for the quality control and standardization of LHQW capsules.

Li Q., Lan T., He S., Chen W., Li X., Zhang W., Liu Y., Zhang Q., Chen X., Han Y., Su Z., Zhu D., Guo H.

Lei-gong-gen formula granule (LFG) is a folk prescription derived from Zhuang nationality, the largest ethnic minority among 56 nationalities in China. It consists of three herbs, namely Eclipta prostrata (L.) L., Smilax glabra Roxb, and Centella asiatica (L.) Urb. It has been widely used as health protection tea for hundreds of years to prevent hypertension in Guangxi Zhuang Autonomous Region. The purpose of this study is to validate the antihypertensive effect of LFG on the spontaneously hypertensive rat (SHR) model, and to further identify the effective components and anti-hypertension mechanism of LFG. The effects of LFG on blood pressure, body weight, and heart rate were investigated in vivo using the SHR model. The levels of NO, ANG II, and ET-1 in the serum were measured, and pathological changes in the heart were examined by H&E staining. The main active components of LFG, their corresponding targets, and hypertension associated pathways were discerned through network pharmacology analysis based on the Traditional Chinese Medicine Systems Pharmacology (TCMSP), Traditional Chinese Medicine Integrated Database (TCMID), and the Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM). Then the predicted results were further verified by molecular biology experiments such as RT-qPCR and western blot. Additionally, the potential active compounds were predicted by molecular docking technology, and the chemical constituents of LFG were analyzed and identified by UPLC-QTOF/MS technology. Finally, an in vitro assay was performed to investigate the protective effects of potential active compounds against hydrogen peroxide (H2O2) induced oxidative damage in human umbilical vein endothelial cells (HUVEC). LFG could effectively reduce blood pressure and increase serum NO content in SHR model. Histological results showed that LFG could ameliorate pathological changes such as cardiac hypertrophy and interstitial inflammation. From network pharmacology analysis, 53 candidate active compounds of LFG were collected, which linked to 765 potential targets, and 828 hypertension associated targets were retrieved, from which 12 overlapped targets both related to candidate active compounds from LFG and hypertension were screened and used as the potential targets of LFG on antihypertensive effect. The molecular biology experiments of the 12 overlapped targets showed that LFG could upregulate the mRNA and protein expressions of NOS3 and proto-oncogene tyrosine-protein kinase SRC (SRC) in the thoracic aorta. Pathway enrichment analysis showed that the PI3K-AKT signaling pathway was closely related to the expression of NOS3 and SRC. Moreover, western blot results showed that LFG significantly increased the protein expression levels of PI3K and phosphorylated AKT in SHR model, suggesting that LFG may active the PI3K-AKT signaling pathway to decrease hypertension. Molecular docking study further supported that p-hydroxybenzoic acid, cedar acid, shikimic acid, salicylic acid, nicotinic acid, linalool, and histidine can be well binding with NOS3, SRC, PI3K, and AKT. UPLC-QTOF/MS analysis confirmed that p-hydroxybenzoic acid, shikimic acid, salicylic acid, and nicotinic acid existed in LFG. Pre-treatment of HUVEC with nicotinic acid could alleviate the effect on cell viability induced by H2O2 and increase the NO level in cell supernatants. LFG can reduce the blood pressure in SHR model, which might be attributed to increasing the NO level in serum for promoting vasodilation via upregulating SRC expression level and activating the PI3K-AKT-NOS3 signaling pathway. Nicotinic acid might be the potential compound for LFG antihypertensive effect.

Li R., Xia Z., Li B., Tian Y., Zhang G., Li M., Dong J.

Ginkgo biloba L. has always been a popular area of research due to its various active ingredients and pharmacological effects. Ginkgo biloba is rich in ginkgo flavonoids, ginkgolides, and ginkgolic acid, with anti-inflammation, antioxidation, neuroprotection, anti-platelet agglutination, hypolipidemic effect, anti-cancer, and anti-radiation properties. There are many methods to extract and separate the active components of ginkgo. Among them, supercritical carbon dioxide fluid extraction (SFE-CO2) is known for its green, clean, and environment-friendly properties. In this paper, the pharmacological activities, the active components, and structures of different parts of ginkgo, the extraction methods of its effective ingredients, and the application of the SFE-CO2 method for the extraction and separation of active ingredients in Ginkgo biloba from leaves, seeds, pollen, and roots were reviewed, in order to make best use of ginkgo resources, and provide support and references for the development of SFE-CO2 of active components from Ginkgo biloba.

Liu M., Cai M., Ding P.

Most of traditional Chinese herbal medicine (TCHM) substances come from medicinal plants, among which oligosaccharides have gradually attracted widespread attention at home and abroad due to their important biological activities and great medicinal potential. Numerous in vitro and in vivo experiments exhibited that oligosaccharides possess various activities, such as antitumor, anti-oxidation, modulate the gut microflora, anti-inflammatory, anti-infection, and immune-regulatory activities. Generally, biological activities are closely related to chemical structures, including molecular weight, monosaccharide composition, glycosidic bond connection, etc. The structural analysis of oligosaccharides is an important basis for studying their structure–activity relationship, but the structural diversity and complexity of carbohydrate compounds limit the study of oligosaccharides activities. Understanding the structures and biological functions of oligosaccharides is important for the development of new bioactive substances with natural oligosaccharides. This review provides a systematic introduction of the current knowledge of the chemical structures and biological activities of oligosaccharides. Most importantly, the reported chemical characteristics and biological activities of the famous TCHM oligosaccharides were briefly summarized, including Morinda officinalis, Rehmannia glutinosa, Arctium lappa, Polygala tenuifolia, Panax ginseng, Lycium barbarum and Astragalus membranaceus. TCHM oligosaccharides play an important role in nutrition, health care, disease diagnosis and prevention as well as have broad application prospects in the field of medicine.

Li N., Xie L., Yang N., Sun G., Liu H., Bi C., Duan J., Yuan Y., Yu H., Xu Y., Li Y.

Introduction Epimedium koreanum Nakai (EKN), is a well-known Chinese herbal medicine for the treatment of osteoporosis, immunosuppression, tumours and cardiovascular diseases. Comprehensive component identification is essential for elucidation of its pharmacological mechanism and quality control. However, its complex chemical composition has caused certain difficulties in the analysis of this traditional Chinese medicine (TCM). Therefore, there is an urgent need to establish a method for rapid classification and identification of EKN chemical components. Objective To establish a method for rapid classification and identification of the main components of flavonoids, organic acids and alkaloids in EKN. Methods The samples were analysed by ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and data post-processing techniques. The UPLC system used a BEH C18 column to separate the total extract of EKN. The mobile phase consisted of 0.1% formic acid in water and acetonitrile, and the EKN extract was analysed by gradient elution at a flow rate of 0.4 mL/min. In both the positive and negative ion modes, the fragment information was obtained and compared with those of the characteristic fragmentations and neutral losses described in the literature to quickly identify the target compounds. Results Finally, we successfully screened out 51 chemical components, including 40 flavonoids, nine organic acids, and two alkaloids. Conclusion The established method not only comprehensively analysed the chemical compositions of EKN, solved the difficult problems of analysis and identification of the complex chemical compositions of the TCM, but also further promoted the development of the application of chemical compositions of TCM.

Nijat D., Abdulla R., Liu G., Luo Y., Aisa H.A.

• This was the first study that was conducted by UHPLC-Quadrupole-Orbitrap-HRMS and HPLC-tri-quadrupole-LIT-MS for Meiguihua oral solution (MOS). • Totally 46 compounds (21 flavonoids and 25 tannins) were firstly detected in MOS, and among them 14 polyphenols were identified for the first time in this herb. • This study provides the simultaneous quantification method for ten compounds including hyperoside and isoquercitrin (isomers). Meiguihua oral solution (MOS), a classical Chinese medicinal formula, was approved by the China Food and Drug Administration for production. However, the quality evaluation of MOS has not been reported. In this present study, qualitative and quantitative analysis of MOS were conducted by ultra high performance liquid chromatography coupled to quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-orbitrap-HRMS) and high performance liquid chromatography-tri-quadrupole linear ion trap mass spectrometry (HPLC-tri-Q-LIT-MS). Totally 46 phenolic compounds (21 flavonoids and 25 tannins) were identified in the MOS, among them 14 polyphenols were not reported in raw plant materials of MOS. The simultaneous quantification of ten compounds including gallic acid, quercetin-3- O -sophoroside, ellagic acid, sophoraflavonoloside, hyperoside, isoquercitrin, avicularin, astragalin, quercitrin and juglanin, which were completed in 16 min in the negative electrospray ionization (ESI) mode under multiple reaction monitoring (MRM) method. Linearity was reached fine determination coefficient (r 2 > 0.9995). Precisions, repeatability, stability (inter-day and intra-day), and recovery were validated and the relative standard deviations (RSD) were less than 2.9%, 4.7%, 3.6% and 1.79%, respectively. This result proved the high sensitivity and efficiency of the method. The quantitative and qualitative analysis of MOS would provide the substantial basis for further quality control and medicinal values.

Wang J., Zhao J., Yan Y., Liu D., Wang C., Wang H.

BACKGROUND Controlling the blood glucose level is an effective method to reduce type 2 diabetes and prevent diabetes-related complications. Ursolic acid is a plant extract that can reduce postprandial hyperglycemia effectively. This study aimed to explore the inhibitory effect and interaction mechanism of ursolic acid against α-amylase and α-glucosidase. RESULTS In this study, the effect of ursolic acid on glycosidase was studied in vitro, in vivo, and in silico. The half-maximal inhibitory concentration (IC50) of ursolic acid on α-amylase and α-glucosidase was 0.482 ± 0.12 mg mL−1 and 0.213 ± 0.042 mg mL−1, respectively. The results of enzymatic kinetics showed that ursolic acid inhibited α-amylase and α-glucosidase activity in a non-competitive manner. The fluorescence spectrum showed that the combination of ursolic acid and glycosidase caused the intrinsic fluorescence quenching of glycosidase. The observation of starch granules revealed that the activity of α-amylase was inhibited and the hydrolysis of starch granules was prevented in the presence of ursolic acid. Molecular docking results showed that ursolic acid bound to the inactive site of α-amylase and α-glucosidase through the formation of ursolic acid–glucosidase complex. Ursolic acid interacted with α-amylase and α-glucosidase mainly through hydrogen bonding. The postprandial hypoglycemic effect of ursolic acid in C57BL/6J mice showed that the high concentration of ursolic acid could quickly reduce postprandial blood glucose level. CONCLUSION Ursolic acid can be considered as a natural ingredient in functional foods to control postprandial blood glucose levels and prevent diabetes by delaying the digestion of starch in foods. © 2019 Society of Chemical Industry

Han X., Wang S., Yang X., Li T., Zhao H., Zhou L., Zhao L., Bao Y., Meng X.

Patrinia villosa (Thunb.) Juss. (PVJ) is described as pungent, bitter and slightly cold in Chinese medicine, and is associated with the large intestine, stomach and liver meridians. The preliminary experiments of our research team proved that PVJ total flavonoids have excellent inhibitory effects on liver cancer cells. The present experiment uses the UPLC-Q-TOF-MS technology and serum pharmacochemistry methods to analyze the chemical components in vitro and in vivo of PVJ antiliver tumors. A total of 14 chemical components were identified in the total flavonoids extract of PVJ, and it is mainly composed of flavonoids, flavonones, flavonols and phenolic acids. At the same time, seven prototypical components and seven metabolic components were detected in the drug-containing plasma. Hydrocaffeate and scutellarein are the phase I metabolites of caffeic acid and scutellarin, respectively. Sulfated apigenin, sulfated luteolin, sulfated kaempferol and methylated kaempferol are the II phase metabolites of apigenin, luteolin, kaempferol, respectively. The experiment provides a reference for the research and development of antitumor drug candidates, and provides a basis for revealing the bioactive components of PVJ and the antitumor mechanism.

Mlala S., Oyedeji A.O., Gondwe M., Oyedeji O.O.

Non-communicable diseases (NCDs) such as cancer, diabetes, and chronic respiratory and cardiovascular diseases continue to be threatening and deadly to human kind. Resistance to and side effects of known drugs for treatment further increase the threat, while at the same time leaving scientists to search for alternative sources from nature, especially from plants. Pentacyclic triterpenoids (PT) from medicinal plants have been identified as one class of secondary metabolites that could play a critical role in the treatment and management of several NCDs. One of such PT is ursolic acid (UA, 3 β-hydroxy-urs-12-en-28-oic acid), which possesses important biological effects, including anti-inflammatory, anticancer, antidiabetic, antioxidant and antibacterial effects, but its bioavailability and solubility limits its clinical application. Mimusops caffra, Ilex paraguarieni, and Glechoma hederacea, have been reported as major sources of UA. The chemistry of UA has been studied extensively based on the literature, with modifications mostly having been made at positions C-3 (hydroxyl), C12-C13 (double bonds) and C-28 (carboxylic acid), leading to several UA derivatives (esters, amides, oxadiazole quinolone, etc.) with enhanced potency, bioavailability and water solubility. This article comprehensively reviews the information that has become available over the last decade with respect to the sources, chemistry, biological potency and clinical trials of UA and its derivatives as potential therapeutic agents, with a focus on addressing NCDs.

Zhao Y., You X., Jiang H., Zou G., Wang B.

Leontopodium leontopodioides (Willd.) Beauv. is used therapeutically to prevent numerous diseases. Historically, L. leontopodioides extracts have been used to treat influenza infections, bronchitis, acute and chronic nephritis, proteinuria, hematuria, and diabetes. However, the bioactive compounds that are responsible for the associated therapeutic effects have not yet been characterized. In this study, high-performance liquid chromatography was utilized to study the anti-inflammatory properties of L. leontopodioides through analysis of spectrum-effect relationships. The bioactive compounds that correlated with anti-inflammatory activities were partially identified. Following aqueous extraction, a variety of different polar organic solvents including petrol ether extracts, ethyl acetate extracts, n-butanol extracts, and residual aqueous extracts were successfully isolated from L. leontopodioides. These extracts were analyzed using high-performance liquid chromatography to generate HPLC fingerprints. A total of 32 common peaks were selected following a similarity analysis (SA). The spectrum-effect relationship was subsequently studied and inflammatory factors were identified following acute inflammatory experiments. The results revealed that the main peaks associated with anti-inflammatory activities were x1, x3, x4, x13, x14, x16 for interleukin-1 (IL-1), x5, x8, x9, x18, x26, x27, x30, x31, x32 for interleukin-6 (IL-6), and x28 and x29 for leukotriene B4 (LTB4). Following analysis of HPLC data, peaks x9 and x14 were identified as chlorogenic acid and ferulic acid, respectively. The current study utilized HPLC and pharmacological analyses to formulate a spectrum-effect relationship and identify bioactive compounds in L. leontopodioides.

Xiao Y., Lv L., Gou P., Xie H.

Eight previously undescribed acyl atractyligenin and carboxyatractyligenin glycosides were isolated from whole Antennaria rosea subsp. confinis (Greene) R. J. Bayer (Compositae) [syn. Leontopodium leontopodioides (Willd.) Beauv. (Asteraceae)] plants and their structures were determined by spectroscopic and chemical methods. The compounds were trivially named leontopodiosides F-M. Seven of the compounds showed potent in vitro inhibitory activity toward pancreatic lipase with IC50 values ranging from 3.4 to 52.5 μM, suggesting that they participate in the previously observed effect this plant has in reducing triglyceride absorption in rats.

Gou P., Xiao Y., Lv L., Xie H.

Three new glucosides of hydroquinone, monoterpene, and megastigmane, benzyl 2,5-dihydroxybenzoate 5-O-β-d-glucopyranoside (isotrichocarpin, 1), (2S,3R)-3,7-dimethyl-6-octene-1,2,3-triol 2-O-β-d-glucopyranoside (leontopodioside D, 4), and (6R,7R,8R,9S)-6,9-epoxy-7,8-dihydroxymegastigman-4-en-3-one 8-O-β-d-glucopyranoside (leontopodioside E, 7) were isolated from the whole herbs of Leontopodium leontopodioides (Willd.) Beauv. (Asteraceae), along with nebrodenside A (2), pungenin (3), betulalbuside A (5), geranyl O-β-d-glucopyranoside (6), and 3β-hydroxy-β-ionone 3-O-β-d-glucopyranoside (8). Their structure were determined by spectroscopic and chemical methods. All the known compounds were reported from this species for the first time. Compounds 2-6 showed potent in vitro pancreatic lipase inhibitory activity, suggesting their participation in the reductive effect of the herbs on triglyceride absorption.

Zhang Y., Yang Y., Ruan J., Chen Q., Li J., Guo Y., Han L., Wang T.

A phytochemical investigation to obtain new triglyceride (TG) accumulation inhibitors resulted in the isolation of six new isobenzofuranones, leontopodiols A (1), B (2), leontopodiosides C (3), D (4), E (5), F (6), together with three known ones (7-9) from the aerial parts of Leontopodium leontopodioides (Willd.) Beauv. The structures of these isolates were identified by routine NMR experiments, optical rotation determination, electronic circular dichroism (ECD) calculation, along with chemical reaction. Moreover, compounds 1, 2, 5, and 7-9 displayed TG accumulation inhibitory effects on HepG2 cells.

Gao Y., Rao H., Mao L., Ma Q.

The essential oil (EO) from the aerial parts of Leontopodium leontopodioides (Willd.) Beauverd was obtained by hydrodistillation and analysed by GC-FID and GC-MS. Sixty-five compounds were identified which represent 96.2% of the total composition of the EO. The major components of the EO were palmitic acid (11.6%), n-pentadecanal (5.7%), linalool (3.8%), β-ionone (3.3%), hexahydrofarnesyl acetone (3.2%), bisabolone (3.2%) and β-caryophyllene (3.2%). The EO exhibited an excellent antibacterial activity against Staphylococcus aureus and Bacillus subtilis according to the MIC values tested by micro-dilution method. It also exhibited a significant cytotoxicity against HepG2 and MCF-7 cell lines with the IC50 values of 67.44 and 70.49 μg/mL according to the MTT assay. However, the antioxidant activity test revealed that the EO exhibited a weak DPPH radical-scavenging activity. In conclusion, the EO of L. leontopodioides could be regarded as a bioactive natural product and deserves further study for its potential therapeutic effects.

Chen Q., Li J., Ruan J., Qu L., Wei H., Ma X., Zhang Y., Wang T.

Five new compounds, leontoaerialosides A (1), B (2), C (3), D (4), and E (5) were obtained from the 70% EtOH extract of the whole plants of Leontopodium leontopodioides (Wild.) Beauv. Their structures were elucidated by chemical and spectroscopic methods. Moreover, compounds 4 and 5 showed significant effects on stimulating the hepatic glucose uptake in HepG2 cells.

Liu W., Lu D., Jia S., Yang Y., Meng F., Du Y., Yang Y., Yuan L., Nan Y.

Benitez D.A., Galvez A.K., Sesenes R.L., Arteaga A.M., Rodriguez J.G.

The effect of Annona muricata leaves extract as an inhibitor of green corrosion of aluminum was investigated 20% cane ethanol and 80% gasoline blend solution for 91 days of immersion. It was evaluated using gravimetric and electrochemical techniques, such as electrochemical impedance spectroscopy (EIS), linear polarization resistance (LPR), and electrochemical noise (EN). The studies revealed that Annona muricata extract is an efficient corrosion inhibitor, reaching its highest efficiency at concentrations of 20 and 25 ppm. This decrease in the corrosion rate is because the inhibitor adsorbs to the aluminum surface by physisorption according to the Langmuir adsorption isotherm and demonstrated a more notable protective effect Fourier transform infrared spectroscopy (FTIR) and gas chromatography coupled with mass spectrometry (GC–MS) analyzes were performed, which showed the presence of phenolic compounds, lauric acid, palmitic acid, linoleic acid, vitamin E, and other compounds with antioxidant properties. The morphology of the aluminum surface was examined by scanning electron microscopy (SEM), showing the existence of a protective layer of Annona muricata extract that reduces pitting corrosion susceptibility of aluminum.