Qualitative and Quantitative Analysis of the Major Bioactive Components of Juniperus chinensis L. Using LC-QTOF-MS and LC-MSMS and Investigation of Antibacterial Activity against Pathogenic Bacteria

Plants in the genus Juniperus have been reported to produce a variety of chemical components, such as coumarins, flavonoids, lignans, sterols, and terpenoids. Here, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) were applied to qualitatively and quantitatively analyze the major bioactive components in an ethanolic crude extract from the leaves of Juniperus chinensis L., which grows naturally in Korea. In addition, the antibacterial activity of the crude extract against pathogenic bacteria was investigated. Using LC-QTOF-MS analysis, we identified ten compounds, of which six were confirmed to be flavonoid and lignan-based components as the major bioactive components, i.e., isoquercetin, quercetin-3-O-α-l-rhamnoside, hinokiflavone, amentoflavone, podocarpusflavone A, and matairesinoside. Among them, a quantitative analysis performed using LC-MS/MS revealed that the levels of quercetin-3-O-α-l-rhamnoside and amentoflavone in the crude extract were 203.78 and 69.84 mg/g, respectively. Furthermore, the crude extract exhibited potential antibacterial activity against 10 pathogenic bacteria, with the highest antibacterial activity detected against Bordetella pertussis. Thus, further studies of the leaf extract of J. chinensis L. must be carried out to correlate the compounds present in the extract with the antibacterial activity and elucidate the mechanisms of action of this extract against bacteria.

[1]  Z. Ali,et al.  Simultaneous determination and characterization of flavonoids, sesquiterpene lactone, and other phenolics from Centaurea benedicta and dietary supplements using UHPLC-PDA-MS and LC-DAD-QToF. , 2022, Journal of pharmaceutical and biomedical analysis.

[2]  A. Almansour,et al.  Bioactive compounds from Euphorbia schimperiana with cytotoxic and antibacterial activities , 2021 .

[3]  Jinhua Tao,et al.  Quality Evaluation of Artemisia capillaris Thunb. Based on Qualitative Analysis of the HPLC Fingerprint and UFLC-Q-TOF-MS/MS Combined with Quantitative Analysis of Multicomponents , 2021, Journal of analytical methods in chemistry.

[4]  Ganjun Yuan,et al.  Antibacterial activity and mechanism of plant flavonoids to gram-positive bacteria predicted from their lipophilicities , 2021, Scientific Reports.

[5]  Y. Rizal,et al.  Identification of bioactive compounds in gambier (Uncaria gambir) liquid by-product in West Sumatra, Indonesia , 2021 .

[6]  F. Lamari,et al.  Metabolomic fingerprinting and genetic discrimination of four Hypericum taxa from Greece. , 2020, Phytochemistry.

[7]  Yuting Chen,et al.  UHPLC‐Q‐TOF‐MS/MS method based on four‐step strategy for metabolites of hinokiflavone in vivo and in vitro , 2019, Journal of pharmaceutical and biomedical analysis.

[8]  Kumar Ganesan,et al.  Isoquercetin upregulates antioxidant genes, suppresses inflammatory cytokines and regulates AMPK pathway in streptozotocin-induced diabetic rats. , 2019, Chemico-biological interactions.

[9]  L. Mondello,et al.  Comparative study of the phenolic profile, antioxidant and antimicrobial activities of leaf extracts of five Juniperus L. (Cupressaceae) taxa growing in Turkey , 2018, Natural product research.

[10]  M. Iranshahi,et al.  Antibacterial activity of flavonoids and their structure–activity relationship: An update review , 2018, Phytotherapy research : PTR.

[11]  A. Ammar,et al.  Antibacterial and antioxidant activity of Juniperus thurifera L. leaf extracts growing in East of Algeria , 2018, Veterinary world.

[12]  A. Sureda,et al.  Therapeutic potential of flavonoids in inflammatory bowel disease: A comprehensive review , 2017, World journal of gastroenterology.

[13]  Haiyan Xiang,et al.  Antioxidant profiling of vine tea (Ampelopsis grossedentata): Off-line coupling heart-cutting HSCCC with HPLC-DAD-QTOF-MS/MS. , 2017, Food chemistry.

[14]  Sam F. Y. Li,et al.  A Review on the Phytochemistry, Pharmacology, and Pharmacokinetics of Amentoflavone, a Naturally-Occurring Biflavonoid , 2017, Molecules.

[15]  Xu Zhou,et al.  Screening of minor bioactive compounds from herbal medicines by in silico docking and the trace peak exposure methods. , 2016, Journal of chromatography. A.

[16]  G. Wang,et al.  Rapid Screening and Structural Characterization of Antioxidants from the Extract of Selaginella doederleinii Hieron with DPPH-UPLC-Q-TOF/MS Method , 2015, International journal of analytical chemistry.

[17]  M. Shivakumar,et al.  Antioxidant compound Quercetin-3-O-α-L-rhamnoside (1→6)-β-D-glucose (Rutin) isolated from ethyl acetate leaf extracts of Memecylon edule Roxb (Melastamataceae) , 2015 .

[18]  Wei Zhang,et al.  Research on the bioactivity of isoquercetin extracted from marestail on bladder cancer EJ cell and the mechanism of its occurrence , 2015, Artificial cells, nanomedicine, and biotechnology.

[19]  Xiaoqing Chen,et al.  Antibacterial activities of flavonoids: structure-activity relationship and mechanism. , 2014, Current medicinal chemistry.

[20]  V. Bagla,et al.  Antimicrobial activity, toxicity and selectivity index of two biflavonoids and a flavone isolated from Podocarpus henkelii (Podocarpaceae) leaves , 2014, BMC Complementary and Alternative Medicine.

[21]  N. Allouche,et al.  In vitro anti-diabetic, anti-obesity and antioxidant proprieties of Juniperus phoenicea L. leaves from Tunisia , 2014 .

[22]  Hong-Guang Jin,et al.  Amentoflavone Stimulates Mitochondrial Dysfunction and Induces Apoptotic Cell Death in Candida albicans , 2012, Mycopathologia.

[23]  C. Lee,et al.  Protective effects of amentoflavone on Lamin A-dependent UVB-induced nuclear aberration in normal human fibroblasts. , 2011, Bioorganic & medicinal chemistry letters.

[24]  N. Miceli,et al.  Antioxidant and antimicrobial activities of branches extracts of five Juniperus species from Turkey , 2011, Pharmaceutical biology.

[25]  J. Ragle,et al.  IUCN Red List of Threatened Species , 2010 .

[26]  J. Bouajila,et al.  Chemical composition and antimicrobial and antioxidant activities of essential oils and various extracts of Juniperus phoenicea L. (Cupressacees). , 2009, Journal of food science.

[27]  M. Rahimizadeh,et al.  Chemical and Antimicrobial Studies of Juniperus excelsa subsp. excelsa and Juniperus excelsa subsp. polycarpos Essential Oils , 2008 .

[28]  Yong Sup Lee,et al.  An update on bioactive plant lignans. , 2005, Natural product reports.

[29]  Andrew J Lamb,et al.  Antimicrobial activity of flavonoids , 2005, International Journal of Antimicrobial Agents.

[30]  L. Nahar,et al.  Biological Activity of Lignans from the Seeds of Centaurea scabiosa , 2003 .

[31]  M. Vaneechoutte,et al.  Susceptibility Testing of Fluconazole by the NCCLS Broth Macrodilution Method, E-Test, and Disk Diffusion for Application in the Routine Laboratory , 2002, Journal of Clinical Microbiology.

[32]  L. Thompson,et al.  Antioxidant activity of the flaxseed lignan secoisolariciresinol diglycoside and its mammalian lignan metabolites enterodiol and enterolactone , 1999, Molecular and Cellular Biochemistry.

[33]  Yuh-meei Lin,et al.  Hinokiflavone, a cytotoxic principle from Rhus succedanea and the cytotoxicity of the related biflavonoids. , 1989, Planta medica.

[34]  G. Towers,et al.  Biological activities of lignans , 1984 .

[35]  M. Okigawa,et al.  Biflavones in Selaginella species , 1971 .

[36]  N. Kawano,et al.  Studies on Bisflavones in the Leaves of Podocarpus macrophylla and P. nagi , 1969 .

[37]  Miguel Ángel Martínez,et al.  Antifungal and Antibacterial Activities of Araucaria araucana (Mol.) K. Koch Heartwood Lignans , 2006, Zeitschrift fur Naturforschung. C, Journal of biosciences.

[38]  Artur M. S. Silva,et al.  The chemical composition of the Juniperus genus (1970-2004) , 2005 .