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The genus Fuchsia is generally used in herbal preparations to treat conditions caused by microorganisms. Based on the popular use of this type of plants, the objective of this study was to obtain sequential extracts of increasing polarity from the branches of Fuchsia lycioides by maceration at room temperature and by the Soxhlet method at 60ºC, to later evaluate the antifungal capacity of the extracts against different clinical isolates of the Candida genus. The ethyl acetate extract exhibited strong anti-fungal activity, selectively inhibiting C. albicans strains with MIC and CMF values of 10 and 15 µg/mL, respectively; comparable with the drug itraconazole®. The analysis of the extract by GC-MS showed a high concentration of terpenoids (mainly phytol) and phenylpropanoids (mainly cinnamic acid), possibly responsible for the antifungal activity of the ethyl acetate extract of F. lycioides.
El género Fuchsia se usa generalmente en preparaciones de hierbas para tratar afecciones provocadas por microorganismos. En base al uso popular de este tipo de plantas, el objetivo de este estudio fue obtener los extractos secuenciales de polaridad creciente de las ramas de Fuchsia lycioides por maceración a temperatura ambiente y por el método Soxhlet a 60ºC, para luego evaluar la capacidad antifúngica de los extractos frente a diferentes aislados clínicos del genero Candida. El extracto de acetato de etilo exhibió una fuerte actividad antifúngica inhibiendo en forma selectiva las cepas de C. albicans con valores de CMI y de CMF de 10 y 15 µg/mL, respectivamente; comparables con el fármaco itraconazol®. El análisis del extracto por CG-EM mostró una alta concentración de terpenoides (principalmente fitol) y fenilpropanoides (principalmente ácido cinámico), posibles responsables de la actividad antifúngica del extracto de acetato de etilo de F. lycioides.
Subject(s)
Candida albicans/drug effects , Plant Extracts/pharmacology , Onagraceae/chemistry , Antifungal Agents/pharmacology , Phenylpropionates/analysis , Temperature , Terpenes/analysis , Plant Extracts/chemistry , Microbial Sensitivity Tests , Gas Chromatography-Mass Spectrometry , Antifungal Agents/chemistryABSTRACT
Phenylpropanoids are one of the major chemical constituents in Zanthoxylum species. They include simple phenylpropanoids, coumarins, and lignans and possess anti-tumor, anti-inflammatory, anti-platelet aggregation, anti-bacterial, anti-viral, insecticidal, and antifeedant activities. This review summarizes the chemical constituents and pharmacological activities from the Zanthoxylum plants in hopes of providing reference for the research and application of phenylpropanoids from this genus.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Coumarins/pharmacology , Lignans , Plant Extracts , ZanthoxylumABSTRACT
Objective:To rapidly identify the chemical constituents in Xiao Chengqitang by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap-MS). Method:The method was established by the Waters CORTECS T3 column (2.1 mm×150 mm, 1.6 μm), mobile phase was methanol (A)-0.1% formic acid aqueous solution (B) for gradient elution (0-5 min, 3%-21%A; 5-20 min, 21%-36%A; 20-32 min, 36%-50%A; 32-42 min, 50%-62%A; 42-50 min, 62%-85%A; 50-60 min, 85%-95%A), the flow rate was 0.2 mL·min<sup>-1</sup>, and the column temperature was 30 ℃. UPLC-Q-Orbitrap-MS was operated in positive and negative ion modes, the scanning range was 100-1 200 with mode of Full MS/dd-MS<sup>2</sup>, and the collision energies were 20, 40 eV. The compounds were identified by comparing with reference substances and combining with literature reports and MS database information. Result:A total of 123 components were identified in Xiao Chengqitang, including 33 flavonoids, 25 anthraquinones and anthrones, 23 phenylpropanoids, 15 tannins, 10 nitrogen-containing components and 17 other components. Among them, 32 components were determined by reference substances. Conclusion:The material basis of Xiao Chengqitang is flavonoids, anthraquinones and anthrones, phenylpropanoids, which is derived from Aurantii Fructus Immaturus,<italic> </italic>Rhei Radix et Rhizoma and Magnoliae Officinalis Cortex, respectively.
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Objective:To rapidly identify the chemical constituents of Chaishi Tuire granules by ultra-performance liquid chromatography-electrospray/quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS). Method:Chromatographic separation was conducted on a Phenomenex<sup>®</sup> Luna omega C<sub>18</sub> column (2.1 mm×100 mm, 1.6 μm) with 0.1% formic acid aqueous solution (A)-acetonitrile (B) as the mobile phases for gradient elution (0-20 min, 5%-40%B; 20-40 min, 40%-95%B; 40-43 min, 95%B), the flow rate was set at 0.3 mL·min<sup>-1</sup>. MS data were collected in positive and negative ion modes, the scanning range was <italic>m</italic>/<italic>z</italic> 150-1 500 and electrospray ionization (ESI) was employed. The chemical constituents of Chaishi Tuire granules were identified by comparing with the retention time and the mass data of the reference substances, as well as the accurate mass, MS/MS fragment ions, mass spectrometry databases (PubChem, MassBank, ChemicalBook and others) and related literature. Result:A total of 85 chemical constituents were identified, including 28 flavonoids, 24 phenylpropanoids, 11 terpenoids, 10 alkaloids, 4 quinones, and 8 others. Among them, 19 constituents derived from Lonicerae Japonicae Flos, 14 constituents derived from Scutellariae Radix, 10 constituents derived from Isatidis Radix, 9 constituents derived from Taraxaci Herba, 9 constituents derived from Forsythiae Fructus, 4 constituents derived from Bupleuri Radix, 4 constituents derived from Anemarrhenae Rhizoma, and 4 constituents derived from Rhei Radix et Rhizoma. Conclusion:Chaishi Tuire granules is rich in phytochemicals, which are derived from many of traditional Chinese medicines. This study can lay a foundation for the quality control, material basis and <italic>in vivo</italic> metabolic analysis of this preparation.
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Objective:To systematically analyze the chemical constituents of Qizhi Jiangtang capsules by ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry (UPLC-QE-Orbitrap-MS). Method:Analysis was conducted on a ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) with acetonitrile (A)-water (B) as the mobile phase for gradient elution (0-13 min, 1%-25%A; 13-21 min, 25%-35%A; 21-28 min, 35%-85%A; 28-30 min, 85%-100%A; 30-32 min, 100%-1%A). The flow rate was 0.2 mL·min<sup>-1</sup>, the column temperature was 30 ℃, and the volume of sample injection was 3 μL. Electrospray ionization (ESI) was used to collect data in the negative and positive ion modes with the scanning range of <italic>m</italic>/<italic>z</italic> 100-1 500. Meanwhile, a variety of MS analytic methods were used, including comparing with the information of control substances, self-built compounds database and literature references, diagnostic ion filtering, Compound Discoverer 3.0 software, for identification of the chemical components. Result:Based on the above strategy, a total of 52 compounds were identified in Qizhi Jiangtang capsules, and the sources of these compounds were identified. Amino acids were mainly derived from Hirudo, phenylpropanoids were derived from Astragali Radix and Rehmanniae Radix, iridoid glycosides were derived from Rehmanniae Radix, coumarins and triterpenes were derived from Astragali Radix, flavonoids were from Astragali Radix and Polygonati Rhizoma. Conclusion:The established UPLC-QE-Orbitrap-MS analytical method can comprehensively and rapidly analyze and identify of the chemical constituents in Qizhi Jiangtang capsules. Many of the ingredients have been proved by modern pharmacological studies to have the effect of improving related symptoms of diabetes and its complications, reflecting the characteristics of synergistic action of multiple components in Qizhi Jiangtang capsules. This study can provide reference for the further research on the pharmacodynamic material basis and the quality control of Qizhi Jiangtang capsules.
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Objective:An ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was developed to analyze and identify the chemical constituents in <italic>Coptis chinensis</italic> inflorescence. Method:The chromatographic separation was performed on ACQUITY UPLC BEH C<sub>18</sub> column (2.1 mm×100 mm, 1.7 μm) with the mobile phase of 0.1% formic acid aqueous solution (A)-acetonitrile (B) for gradient elution (0-15 min, 10%-22%B; 15-20 min, 22%B; 20-25 min, 22%-44%B; 25-35 min, 44%-50%B; 35-40 min, 50%-60%B; 40-55 min, 60%-85%B), the flow rate was 0.15 mL·min<sup>-1</sup>, the injection volume was 3 μL and the column temperature was 30 ℃. HRMS was equipped with electrospray ionization (ESI) and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan (Full MS/dd-MS<sup>2</sup>). Compound Discoverer 3.0 software combined with mzCloud, mzVault, ChemSpider databases and HRMS database of components in traditional Chinese medicine were used to analyze and identify the collected data by HRMS, based on accurate relative molecular mass, retention time and characteristic ion fragmentation of the compounds, as well as literature information and relevant reference materials. Result:A total of 51 chemical constituents were identified in <italic>C</italic>.<italic> chinensis</italic> inflorescence, including 16 alkaloids, 14 flavonoids, 7 phenylpropanoids, 7 organic acids and 7 others. Among them, 10 components [berberine, palmatine, coptidine, rutin, quercetin, isoquercitrin, chlorogenic acid, cryptochlorogenic acid,<italic> D</italic>-(-) quinic acid and <italic>D</italic>-proline] were unambiguously identified by comparing with reference standards. Conclusion:The established UPLC-Q-Orbitrap HRMS can be used to accurately analyze and identify chemical constituents of <italic>C. chinensis</italic> inflorescence. A total of 41 chemical constituents are reported from <italic>C. chinensis</italic> inflorescence for the first time and 6 alkaloids are found from the <italic>C. chinensis</italic> for the first time. These findings can provide methodological reference and experimental basis for the basic research of quality evaluation and efficacy materials of <italic>C. chinensis</italic> inflorescence, and lay a foundation for its further development and utilization.
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Objective::To study the chemical constituents in Baihe Dihuangtang by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). Method::The separation was performed on an ACQUITY UPLC BEH C18 column (2.1 mm×50 mm, 1.7 μm) by a gradient elution of acetonitrile-0.1%formic acid solution. The flow rate was 0.4 mL·min-1 and the column temperature was 30 ℃, the injection volume was 5 μL. Electrospray ionization was applied and the data were collected via positive and negative ion modes. By using SCIEX OS 1.4 software, the chemical constituents were analyzed based on the retention time, excimer ion peak and fragment ion peak of the compounds, as well as comparison with reference substances and literature data. Result::A total of 49 chemical constituents in Baihe Dihuangtang were identified, including 5 phenolic glycerides, 15 phenylethanoid glycosides, 11 iridoids, 8 lonones, 3 phenylpropanoids, 2 nucleosides, 1 organic acid, and 4 other compounds. Among them, phenolic glycerides belonged to Lilii Bulbus, and other components mainly belonged to Rehmanniae Radix. Four chemical constituents (acteoside, isoacteoside, ferulic acid and caffeic acid) were identified by comparison of reference substances. Conclusion::The established detection method can quickly and accurately analyze the chemical constituents of Baihe Dihuangtang. The information of chemical constituents in Baihe Dihuangtang is comprehensively expounded. The study establishes a foundation for the research of quality control, material foundation of efficacy and the development of compound preparations of Baihe Dihuangtang.
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Objective:To study the water soluble chemical constituents in rhizoma of Acorus tatarinowii and transformation pathway of nucleosides in the process of water extraction. Method:Compounds were isolated and purified by column chromatography on macroporous resin,Sephadex LH-20,ODS and preparative HPLC. Their structures were identified on the basis of physicochemical properties and spectral data. Nucleosides were identified from aqueous extract of A. tatarinowii,and their stability was investigated by HPLC. The possible transformation pathways of nucleosides in aqueous extract of A. tatarinowii were studied by nucleotide addition test. Result:Eleven compounds including four nucleosides,four phenylpropanoids,two alkaloids and a furfural were isolated,and identified as uridine(1),adenine(2),guanosine(3),adenosine(4),5-hydroxymethylfurfural(5),5-(hydroxymethyl)-1H-pyrrole-2-carboxaldehyde(6),(threo)1',2'-dihydroxyasarone(7),(erythro)1',2'-dihydroxyasarone(8),acoraminol A(9),acoraminol B(10),and tatarine A(11). The chromatographic peaks of compounds 1-4 and cytidine were identified from aqueous extract of A. tatarinowii by HPLC. After ultrasonic extraction for 0.5 h,the stability of nucleosides in water was poor. After ultrasonic extraction for 3 h or refluxing extraction for 0.5 h,the stability of nucleosides in water was good. Four transformation pathways including 5'-cytidylic acid→cytidine,uridine monophosphate→uridine,guanosine monophosphate,guanosine and adenosine-5'-monophosphate,adenosine 5'-diphosphate,adenosine 5'-triphosphorate,adenosine,adenine might exist in water extract of A. tatarinowii. Conclusion:Compounds 1-4 and 6 were isolated from the genus Acorus for the first time. These compounds further enriched the chemical constituents of A. tatarinowii. The stability and transformation pathway of nucleosides in A. tatarinowii provides reference data for the analysis of nucleosides in A. tatarinowii and other traditional Chinese medicine.
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Objective: To study the chemical constituents of Changyanning Tablets, a high performance liquid chromatography-quadrupole time of flight mass spectrometry (HPLC-Q-TOF-MS/MS) was established to recognize and classify the ingredients accurately and rapidly. Methods: Agilent ZORBAX Eclipse XDB-C18 chromatographic column (250 mm × 4.6 mm, 5 μm) was employed and the separation was performed with the mobile phase consisting of methanol-0.05% acetic acid aqueous solution. The information of accurate mass and multistage fragment ions were obtained by the monitored simultaneously for positive and negative ions. The main chemical constituents of Changyanning Tablets were identified by high resolution mass spectrometry data, combining with Pubmed, Hmdb, Massbank network database, reference literature and comparing the reference. Results: Fifty-one chemical components were finally identified in this study, including two phenylpropanoids, eight iridoids, 12 flavonoids, four tannins, 23 organic acids, and two other classes. Conclusion: This study comprehensively studies the material basis in Changyanning Tablets, which provides a basis for improving the quality evaluation system of Changyanning Tablets and lays the foundation for elucidating the active components mechanism.
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Three phenylpropanoid glycosides were purified by extensive chromatographic techniques including column chromatography over microporous resin, MCI, diol, sephadex LH-20, reverse phase C_(18) from water-extracts of Sophorae Tonkinensis Radix et Rhizoma. Their planar structures were elucidated by combination of various spectroscopic method, such as IR, UV, MS, and NMR. The absolute configuration of aglycone was determined by quantum chemical calculations. Their structures were elucidated as(8R)-3-(4-hydroxyphenyl)-1-propanol-2-O-β-D-glucopyranoside(1), kalopanaxin D(2),(E)-4-hydroxycinnamyl alcohol 4-O-[2'-O-β-D-apiofuranosyl(1″→2')]-β-D-glucopyranoside(3). Compound 1 was undescribed previously. Compounds 2 and 3 were firstly isolated from Sophora genus.
Subject(s)
Drugs, Chinese Herbal , Ethanol , Glycosides , Rhizome , SophoraABSTRACT
Objective:HPLC for the determination of five components in Descurainiae Semen was established to investigate the change rule of contents of five components in the herb before and after being processed. Method:The contents of quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside(QGG),sinapic acid,quercetin-3-O-β-D-glucopyranoside(QG),isorhamnetin-3-O-β-D-glucopyranoside(IG) and 1,2-di-O-sinapoyl-β-D-glucopyranose(SG) was determined simultaneously by HPLC,the change rule of contents of these components before and after processing and its reasons were analyzed.Waters Symmetry® C18 column(4.6 mm×250 mm,5 μm) was employed,and the mobile phase was acetonitrile(A)-1% acetic acid aqueous solution(B) for gradient elution(0-5 min,5%-10%A;5-15 min,10%-13%A;15-23 min,13%-20%A;23-43 min,20%-25%A;43-46 min,25%A;46-55 min,25%-40%A;55-60 min,40%A).The flow rate was 1 mL·min-1.The detection wavelength was set at 265 nm,the injection volume was 10 μL,and the column temperature was 30℃. Result:Contents of the above five components before processing were 0.114 3%,0.041 6%,0.036 2%,0.022 6% and 0.097 6%;after processing,the contents of these five components turn into 0.107 4%,0.011 3%,0.034 2%,0.021 9% and 0.058 9%;among them,the contents of these five components decreased by 6.04%,72.84%,5.52%,3.10% and 39.65%,respectively. Conclusion:The contents of these five components in Descurainiae Semen is reduced to varying degrees after processing.The contents of phenylpropanoids decrease significantly,while the contents of flavonoid glycosides do not change significantly.
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Objective: To study the chemical constituents of barks of Eucommia ulmoides. Methods: Compounds were isolated by means of normal phase silical gel, reverse phase ODS, Sephadex LH-20 column chromatography, and preparative-HPLC, and their structures were identified on the basis of various spectroscopic analyses, including NMR and HR-ESI-MS. Results: Nine known compounds, including seven flavonoids and two phenylpropanoids, were isolated from the 95% EtOH extract of barks of E. ulmoides. Their structures were identified as licoflavone B (1), 3,5,4’-trihydroxy-7,3’-dimethoxyflavone (2), liquiritigenin (3), formononetine (4), (+)-4’-O-methylglabridin (5), paratocarpin E (6), loureirin C (7), threo-1-(4-hydroxy-3-methoxyphenyl)-1-methoxy-2-{4-[1-formyl- (E)-vinyl]-2-methoxyphenoxy}-3-propanol (8), and diospyrosin (9). Conclusion: Compounds 1-9 are obtained from the genus of Eucommia for the first time.
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Active components of Chinese materia medica (CMM) are the material basis of its efficacy and the keys to realize modernization of CMM. How to improve the content of active ingredients of CMM has become a research hotspot, and microbial transformation is one of its key technologies. The practice has proved that the strains are different, the types of CMM are different, and the transformation effects are different. Therefore, the research on fungal transformation of CMM active ingredients such as terpenes, alkaloids, phenylpropanoids, quinones, steroids, organic acids, etc. is reviewed in this study.
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Xanthii Fructus has been traditionally used for the treatment of rhinitis, rheumatoid arthritis, and eczema. In this study, a high-performance liquid chromatography-photodiode array (HPLC-PDA) method was developed and then used for the simultaneous analysis of eight phenylpropanoids in Xanthii Fructus. The analytical column used for this separation was a SunFire™ C₁₈ column, maintained at 40℃. The mobile phase used was 1.0% acetic acid in distilled water and 1.0% acetic acid in acetonitrile with gradient elution. For identify of each component, the mass spectrometer (MS) was used a Waters triple quadrupole mass spectrometer requipped with electrospray ionization (ESI) source. The HPLC-PDA method showed good linearity: correlation coefficients were ≥ 0.9996. The limits of detection and quantification of the eight compounds were 0.02 – 0.04 and 0.06 – 0.14 µg/mL, respectively. The extraction recoveries ranged from 97.51 to 108.67%. The relative standard deviation values of intra- and inter-day precision were 0.06 – 1.55 and 0.09 – 1.68%, respectively. The validated HPLC-PDA method was applied to simultaneously analyse the amounts of eight phenlypropanoids in Xanthii Fructus.
Subject(s)
Acetic Acid , Arthritis, Rheumatoid , Eczema , Limit of Detection , Methods , Rhinitis , WaterABSTRACT
Plants of the genus Salix L. are abundant in resources, consisting more than 520 species throughout the world. The bark, branches, leaves, and flowers of different Salix species are used in folk medicines with a long history. There are several kinds of chemical components in the Salix plants, such as flavonoids, salicylic acid and its derivatives, phenylpropanoids, terpenoids, anthraquinones and steroids, which have the pharmacological activities of anti-tumor, analgesic, anti-inflammatory, anti-oxidant, hypoglycemic, regulating the nervous system, promoting the lipid dissolution, etc. In the present paper, we summarize the research reports related with plants of the genus Salix from the databases of CNKI and PubMed from 2008 to 2018, and review the research progress of chemical components and pharmacological effects to provide references for the in-depth studies of medicinal active ingredients and their action mechanism on this genus, hoping to promote the further rational development and utilization of the Salix plants.
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Objective: To compare the expression of genes in the leaves of Tussilago farfara that involved in biosynthesis of phenylpropanoids in different developmental stages, and infer the accumulation period of biosynthesis of phenylpropanoids and provide a scientific basis for the resource utilization of leaves of T. farfara. Methods: The Illumina HiSeq2500 highthroughput sequencing method was used to analyze the transcriptome of the leaves of T. farfara in different periods. After obtaining transcriptome data, bioinformatics analysis of gene function annotation was performed to compare the expression of genes related to phenylpropanoid biosynthesis in different periods. Results: A total of 46 793 unigenes were obtained by transcriptome sequencing and the average length was 952.144 8 bp. Among them, 4 774 unigenes were annotated in the public databases NR, Swiss-Prot, eggNOG, GO, and KEGG. According to the assignment of KEGG pathway, 144 unigenes were involved in terpenoid biosynthesis, phenylpropanoid biosynthesis and flavonoids, 65 unigenes were involved in terpenoid biosynthesis, 64 unigenes were involved in phenylpropanoid and 15 unigenes were involved in flavonoids biosynthesis. The enzyme genes involved in the phenylpropanoid biosynthesis were also compared in different development stages, and the results indicated that the expression of PAL, 4CL, HCT, and CCoAOMT, which were closely related to biosynthesis of phenylpropanoids, were highest in September, which means that the contents of these compounds might be highest in September. Conclusion: This study lays the foundation for the biosynthetic pathway and regulation analysis of phenylpropanoids, and provides a scientific basis for the development and the resource utilization of leaves of T. farfara.
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As a traditional Chinese medicine, Pithecellobium clypearia has been commonly used for hundreds of years. Its unilateral preparations are applied for the treatment of upper respiratory tract infections, pharyngitis, laryngitis, acute tonsillitis, acute gastroenteritis and other diseases. The present researches indicate that flavonoids, phenylpropanoids, organic phenolic acids, triterpenes, and steroids are the main chemical constituents of P. clypearia, which have a wide range of pharmacological activities, such as anti-oxidation, anti-virus, inhibition of T lymphocyte proliferation, anti-inflammatory, anti-allergy and so on. According to the domestic and foreign relevant literatures, the chemical composition, quality standards, pharmacological activity, and clinical application of P. clypearia were systematically summarized in this paper, which could provide the reference for the further investigation and development of this herbal plant.
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Aleppo Avens, a traditional Chinese medicine, has been commonly used for the treatment of headache and dizziness, infantile convulsion, hypertension, lumbar and leg pain, irregular menstruation, and so on. However, the underlying mechanisms of the functions remain unclear. Up to the present, terpenoids (monoterpene, sesquiterpene and three terpenoids), tannins and phenylpropyl esters were found as the main chemical composition in Aleppo Avens. Accumulating studies showed that Aleppo Avens possess the pharmacological profile of anticoagulant, antihypertensive, anti-inflammatory, anticancer, and anti-ischemia. The present paper reviewed the researches of active ingredients and pharmacological effects of Aleppo Avens in recent 40 years, and provided the pharmacological basis for the clinical application and further rational development of Aleppo Avens.
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The open silica gel, ODS, and Sephadex LH-20 column chromatography, along with the semi-preparative HPLC was used to isolate and purify the chemical constituents from Murraya euchrestifolia. The structures of the isolates were elucidated by their physiochemical properties, NMR, and MS spectroscopic data, as well as comparison with literature data. Eighteen compounds were isolated from the CH2Cl2 fraction of the 95% aqueous EtOH extract of M. euchrestifolia, and their structures were identified as sakuranetin (1), eriodictyol-7,4'-dimethyl ether (2), isosakuranetin (3), 5-hydroxy-7,4'-dimethoxyflavanone (4), eriodictyol-7-methyl ether (5), lichexanthon (6), 5,6,7-trimethoxycoumarin (7), 5-hydroxy-6,8-dimethoxycoumarin (8), 8-hydroxy-6-methoxy-3-n-pentylisocoumarin (9), ethyl caffeate (10), 4-hydroxy-3,5- dimethoxycinnamic acid ethyl ester (11), methyl 3-(5'-hydroxyprenyl)-coumarate (12), (E)-coniferol (13), β-hydroxypropiovanillone (14), 3-hydroxy-7,8-didehydro-β-ionone (15), 3β-hydroxy-5α, 6α-epoxy-7-megastigmen-9-one (16), grasshopper ketone (17), and 4-hydroxy-3,5-dimethoxybenzaldehyde (18). Compounds 1-15 and 18 were first obtained from the plants of Murraya genus, and compounds 16 and 17 were isolated from M. euchrestifolia for the first time.
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The aim of this study was to develop a simple, sensitive ultra performance liquid chromatography mass spectrometry (UPLC-MS/MS) method for the determination of syringaresinol, N-trans-feruloyltyramine, chelerythrine chloride, sinomenine, coptisine chloride, sanguinarine, chelidonine, magnolflorine, allocryptopine, protopine, farrerol, stylopine and dihydrosanguin-arine in Tong'an injection (TAI), which could be used for the quality control of TAI. The UPLC analysis was performed on Agilent Zorbax SB-Aq column (2.1 mm×150 mm,3.5 μm), with 0.1% formic acid solution (A) -acetonitrile (B) as the mobile phase for gradient elution (0.01-2 min, 5%B; 2-8 min, 5%-30%B; 8-11 min, 30%-95%B; 11-13 min, 95%B; 13-13.1 min, 95%-5%B; 13.1-14 min, 5%B). The flow rate was 0.5 mL•min⁻¹, and the column temperature was 25 ℃; multiple reaction monitoring (MRM) was performed in electrospray ion source positive ion mode for quantitative determination. The calibration curves for the above thirteen compounds showed good linear relationship in corresponding mass concentration range (r>0.999 0). The average recovery rate of the compounds ranged from 95.70% to 104.8%, with RSD of less than 1.9%. The contents of thirteen active components in 10 batches of TAI were 0.021 2-0.029 0, 0.001 7-0.002 3, 0.000 9-0.001 3, 5.952-6.205 2, 0.195 4-0.240 5, 0.002 0-0.002 9, 0.693-0.798 2, 0.069 3-0.078 2, 0.089 29-0.102 9, 0.386 5-0.420 1, 0.014 3-0.015 9, 0.755 3-0.842 1, and 0.008 2-0.011 2 g•L⁻¹ respectively. Methodology validation proved that this method was simple, rapid, sensitive and accurate, which can be used to provide reference for the comprehensive evaluation of TAI quality. The determination results of 10 batches of TAI showed the content of each batch had no significant difference. The results may provide a basis for the quality control of TAI.