Expression of β-glucosidase An-bgl3 from Aspergillus niger for conversion of scopolin / 生物工程学报

Scopoletin is a coumarin compound with various biological activities including detumescence and analgesic, insecticidal, antibacterial and acaricidal effects. However, interference with scopolin and other components often leads to difficulties in purification of scopoletin with low extraction rates from plant resource. In this paper, heterologous expression of the gene encoding β-glucosidase An-bgl3 derived from Aspergillus niger were carried out. The expression product was purified and characterized with further structure-activity relationship between it and β-glucosidase analyzed. Subsequently, its ability for transforming scopolin from plant extract was studied. The results showed that the specific activity of the purified β-glucosidase An-bgl3 was 15.22 IU/mg, the apparent molecular weight was about 120 kDa. The optimum reaction temperature and pH were 55 ℃ and 4.0, respectively. Moreover, 10 mmol/L metal ions Fe2+ and Mn2+ increased the enzyme activity by 1.74-fold and 1.20-fold, respectively. A 10 mmol/L solution containing Tween-20, Tween-80 and Triton X-100 all inhibited the enzyme activity by 30%. The enzyme showed affinity towards scopolin and tolerated 10% methanol and 10% ethanol solution, respectively. The enzyme specifically hydrolyzed scopolin into scopoletin from the extract of Erycibe obtusifolia Benth with a 47.8% increase of scopoletin. This demonstrated that the β-glucosidase An-bgl3 from A. niger shows specificity on scopolin with good activities, thus providing an alternative method for increasing the extraction efficiency of scopoletin from plant material.

Effects of six compounds with different chemical structures on melanogenesis / 中国天然药物

Several chemical compounds can restore pigmentation in vitiligo through mechanisms that vary according to disease etiology. In the present study, we investigated the melanogenic activity of six structurally distinct compounds, namely, scopoletin, kaempferol, chrysin, vitamin D, piperine, and 6-benzylaminopurine. We determined their effectiveness, toxicity, and mechanism of action for stimulating pigmentation in B16F10 melanoma cells and in a zebrafish model. The melanogenic activity of 6-benzylaminopurine, the compound identified as the most potent, was further verified by measuring green fluorescent protein concentration in tyrp1 a: eGFP (tyrosinase-related protein 1) zebrafish and mitfa: eGFP (microphthalmia associated transcription factor) zebrafish and antioxidative activity. All the tested compounds were found to enhance melanogenesis responses both in vivo and in vitro at their respective optimal concentration by increasing melanin content and expression of TYR and MITF. 6-Benzyamino-purine showed the strongest re-pigmentation action at a concentration of 20 μmol·Lin vivo and 100 μmol·Lin vitro, and up-regulated the strong fluorescence expression of green fluorescent protein in tyrp1a: eGFP and mitfa: eGFP zebrafish in vitro. However, its relative anti-oxidative activity was found to be very low. Overall, our results indicated that 6-benzylaminopurine stimulated pigmentation through a direct mechanism, by increasing melanin content via positive regulation of tyrosinase activity in vitro, as well as up-regulating the expression of the green fluorescent protein in transgenic zebrafish in vivo.

Effects of six compounds with different chemical structures on melanogenesis / 中国天然药物

Several chemical compounds can restore pigmentation in vitiligo through mechanisms that vary according to disease etiology. In the present study, we investigated the melanogenic activity of six structurally distinct compounds, namely, scopoletin, kaempferol, chrysin, vitamin D, piperine, and 6-benzylaminopurine. We determined their effectiveness, toxicity, and mechanism of action for stimulating pigmentation in B16F10 melanoma cells and in a zebrafish model. The melanogenic activity of 6-benzylaminopurine, the compound identified as the most potent, was further verified by measuring green fluorescent protein concentration in tyrp1 a: eGFP (tyrosinase-related protein 1) zebrafish and mitfa: eGFP (microphthalmia associated transcription factor) zebrafish and antioxidative activity. All the tested compounds were found to enhance melanogenesis responses both in vivo and in vitro at their respective optimal concentration by increasing melanin content and expression of TYR and MITF. 6-Benzyamino-purine showed the strongest re-pigmentation action at a concentration of 20 μmol·Lin vivo and 100 μmol·Lin vitro, and up-regulated the strong fluorescence expression of green fluorescent protein in tyrp1a: eGFP and mitfa: eGFP zebrafish in vitro. However, its relative anti-oxidative activity was found to be very low. Overall, our results indicated that 6-benzylaminopurine stimulated pigmentation through a direct mechanism, by increasing melanin content via positive regulation of tyrosinase activity in vitro, as well as up-regulating the expression of the green fluorescent protein in transgenic zebrafish in vivo.

Penile Erection Induced by Scoparone from Artemisia capillaris through the Nitric Oxide-Cyclic Guanosine Monophosphate Signaling Pathway

PURPOSE: The objective of this study was to evaluate the relaxant effect of scoparone from Artemisia capillaris on rabbit penile corpus cavernosum smooth muscle (PCCSM) and to elucidate the mechanism of action of scoparone for the treatment of erectile dysfunction (ED). MATERIALS AND METHODS: PCCSM that had been precontracted with phenylephrine was treated with 3 Artemisia herbs (A. princeps, A. capillaris, and A. iwayomogi) and 3 fractions (n-hexane, ethyl acetate, and n-butanol) with different concentrations (0.1, 0.5, 1.0, and 2.0 mg/mL). Four components (esculetin, scopoletin, capillarisin, and scoparone) isolated from A. capillaris were also evaluated. The PCCSM was preincubated with Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME) and 1H-[1,2,4] oxadiazolo [4,3-a]quinoxalin-1-one (ODQ). Cyclic nucleotides in the perfusate were measured by a radioimmunoassay. The interactions of scoparone with udenafil and rolipram were also evaluated. RESULTS: A. capillaris extract relaxed PCCSM in a concentration-dependent manner. Scoparone had the highest relaxant effect on PCCSM among the 4 components (esculetin, scopoletin, capillarisin, and scoparone) isolated from the ethyl acetate fraction. The application of scoparone on PCCSM pretreated with L-NAME and ODQ led to significantly less relaxation. Scoparone also increased the cyclic guanosine monophosphate (cGMP) levels in the perfusate in a concentration-dependent manner. Furthermore, scoparone enhanced udenafil- and rolipram-induced relaxation of the PCCSM. CONCLUSIONS: Scoparone relaxed the PCCSM mainly by activating the nitric oxide-cGMP signaling pathway, and it may be a new promising treatment for ED patients who do not completely respond to udenafil.

Acetylcholinesterase Inhibitors from Angelica polymorpha Stem

Fourteen compounds were isolated from the stem of Angelica polymorpha. On the basis of spectral data, these compounds were identified as isoimperatorin (1), phellopterin (2), bergapten (3), xanthyletin (4), cnidilin (5), geijerine (6), (−)-3'-acetyl hamaudol (7), 7-demethylsuberosine (8), dehydrogeijerin (9), (−)-hamaudol (10), (+)-visamminol (11), divaricatol (12), scopoletin (13), and decursidate (14), respectively. Among them, compounds 4 - 6, 8, 9, 13, and 14 were isolated for the first time from A. polymorpha. Dehydrogeijerin (6) and geijerin (9) were isolated for the first time from genus Angelica. All isolates tested for inhibitory activity against acetylcholinesterae. Compounds 1 to 13 showed acetylcholinesterase inhibitory activity with IC₅₀ values ranging from 1.4 to 37.5 µM.

Determination of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in plants of Erycibe / 中国中药杂志

<p><b>OBJECTIVE</b>An accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.</p><p><b>METHOD</b>Scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.</p><p><b>RESULT</b>Good linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.</p><p><b>CONCLUSION</b>The method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.</p>

HPLC and GC-MS Analysis of Phenolic Substances in Acer tegmentosum

The stem barks, heartwoods, and leaves of Acer tegmentosum (Aceraceae) are widely used in Korea to treat hepatic or cerebral disorders mainly due to alcohol poisoning. This study was aimed to analyze phenolic substances in A. tegmentosum. Quantitative analysis of the three phenolic substances (salidroside, (+)-catechin and scopoletin) was performed by HPLC and the identification of volatile phenolic substances were done by GC-MS. The contents of the three compounds in the three MeOH extracts were higher in the stem bark (salidroside: 80.22 mg/g, (+)-catechin: 23.31 mg/g, and scopoletin: 9.45 mg/g) compared to the heartwoods and leaves. And GC-MS analysis of the stem bark extract demonstrated that p-tyrosol is a main substance of twenty-one compounds identified.

Simultaneous quantitation of artemisinin, arteannuin B, artemisic acid, and scopoletin in mice plasma by HPLC-MS / 中国中药杂志

The objective of this study is to develop a sensitive and reliable high-performance liquid chromatography mass spectrometry (LC-MS) method for simultaneous determination of artemisinin, arteannuin B, artemisic acid, and scopoletin, and study the pharmacokinetics of the four constituents in mouse serum after oral administration of the four components to mice. The analytical column used was Agilent Zorbax SB-C18 (2.1 mm x 150 mm, 5 mm). The mobile phase was acetonitrile: 0.5% acetic acid (60: 40) and the flow rate was 0.3 mL x min(-1). The temperature of the column was 40.0 degrees C. In this condition, we established an analysis method to simultaneously determine the four components. A sensitive and specific liquid chromatography-mass spectrometric (LC-MS) method was developed and validated for the determination of artemisin in derivatives in mice plasma. The method we established has a linear range of 5-3 000 μg x L(-1) with a good sensitivity and specificity for all of the four components. This method is simple, rapid, accurate and suitable for the determination of the content of the four compounds.

Two new phenylpropanoids from Micromelum integerrimum / 中国天然药物

AIM@#To investigate the chemical and bioactive constituents from the stems and leaves of Micromelum integerrimum.@*METHOD@#The chemical constituents were isolated and purified by silica gel, Sephadex LH-20, and HPLC. Their structures were mainly elucidated on the basis of extensive 1D- and 2D-NMR spectroscopy and mass spectrometry. Their cytotoxicity and antimicrobial activities were tested by the SRB and turbidimetric methods, respectively.@*RESULTS@#Two new phenylpropanoids and two known coumarins were obtained, and their structures were identified as microintegerrin A (1), microintegerrin B (2), scopoletin (3), and scopolin (4). All of the compounds were tested for their cytotoxicity against three cancer cell lines (HeLa, A549, and BGC-823) and for antimicrobial activity against the fungus Candida albicans and the bacterium Staphylococcus aureus.@*CONCLUSION@#Two new phenylpropanoids 1 and 2 were isolated and identified from the stems and leaves of M. intgerrimum. None of the compounds showed cytotoxic or antimicrobial activity at the tested concentration of 20 μg·mL(-1).

Scopoletin from Cirsium setidens Increases Melanin Synthesis via CREB Phosphorylation in B16F10 Cells

In this study, we isolated scopoletin from Cirsium setidens Nakai (Compositae) and tested its effects on melanogenesis. Scopoletin was not toxic to cells at concentrations less than 50 microM and increased melanin synthesis in a dose-dependent manner. As melanin synthesis increased, scopoletin stimulated the total tyrosinase activity, the rate-limiting enzyme of melanogenesis. In a cell-free system, however, scopoletin did not increase tyrosinase activity, indicating that scopoletin is not a direct activator of tyrosinase. Furthermore, Western blot analysis showed that scopoletin stimulated the production of microphthalmia-associated transcription factor (MITF) and tyrosinase expression via cAMP response element-binding protein (CREB) phosphorylation in a dose-dependent manner. Based on these results, preclinical and clinical studies are needed to assess the use of scopoletin for the treatment of vitiligo.

Chemical constituents of Rhododendron seniavinii / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Rhododendron seniavinii.</p><p><b>METHOD</b>Compounds were isolated from the aqueous extract of the leaves of R. seniavinii by using Sephadex LH-20, ODS open column chromatography and other means. Their structures were elucidated according to spectral data and physiochemical properties.</p><p><b>RESULT</b>Thirteen compounds were isolated from R. seniavinii and identified as 5-methoxydehydroconiferyl alcohol (1), dehydroconiferyl alcohol (2), (-)-syringaresinol (3), (-)-lyoniresinol (4), (+)-lyoniresinol 3alpha-O-beta-D-glucopyranoside (5), (-)-lyoniresinol 3alpha-O-beta-D-glucopyranoside (6), 3,4,5-trimethoxyphenyl-1-O-beta-D-glucopyranoside (7), nikoenoside (8), 3,5,7-trihydroxychromone-3-0-alpha-L-rhamnopyranoside (9), 3,4,5-trimethoxyphenol (10), scopoletin (11), scopolin (12) and quercitrin (13).</p><p><b>CONCLUSION</b>Compounds 1-12 were obtained from this plant for the first time.</p>

Efecto de Bursera grandiflora sobre el peso corporal y lipemia en ratones obesos / Effect of Bursera grandiflora on body weight and lipemia in obese mice

Obesity and overweight are clearly related to the caloric intake that leads to a chronic systemic inflammation of low degree. Pharmacological therapy is highly recommended for this disease. Within the Burseraceae family there are species used empirically for the treatment of obesity; one of them, Bursera grandiflora, is also reported for exhibiting anti-inflammatory properties. Evaluate the effect produced by B. grandiflora on a mice model of obesity, its toxicity potential, and chemical identification of the major chemical compound. Mice of the C57B1/6 strain were used. All animals were fed for 8 weeks with a hypercaloric diet. Afterwards, during 7 weeks, animals were daily administered with the hidro-alcoholic extract from B. grandiflora or water (control). At the end of the administration period, plasma cholesterol and triglycerides levels were measured. Moreover, toxicity tests were carried out in mice after acute and chronic administering the B. grandiflora extract. Finally, the main compound in the active extract was identified. Animals treated with the B. grandiflora extract showed a greater food consumption and, paradoxically, without increasing the body weight. Moreover, a decrease of the plasma-triglycerides was observed. Toxicological evaluation showed that the extract administration did not produce any death of the experimental animals or modifications on the organs or behavior. The major compound identified in the extract was scopoletin.

El sobrepeso y la obesidad están claramente vinculados con la ingesta calórica que lleva a la instalación crónica de inflamación sistémica de bajo grado. La terapia farmacológica es altamente recomendable para este padecimiento. Dentro de la familia Burseraceae existen especies utilizadas para el tratamiento de la obesidad, de las cuales, Bursera grandiflora es referida también por su efecto anti-inflamatorio. Evaluar el efecto de B. grandiflora en un modelo de obesidad, su potencial toxicológico, e identificación del compuesto mayoritario. Se utilizaron ratones C57Bl/6 alimentados durante 8 semanas con dieta hipercalórica. Posteriormente, durante 7 semanas se administró diariamente un extracto hidroalcohólico de B. grandiflora o agua (control). Al final se cuantificaron colesterol y triglicéridos. Además, se realizaron pruebas de toxicidad en ratones administrando el extracto de B. grandiflora en forma aguda y subcrónica. Finalmente, se identificó el compuesto mayoritario. Los animales tratados con B. grandiflora registraron el mayor consumo de alimento y, paradójicamente, sin mostrar un crecimiento ponderal y con una disminución de los triglicéridos. Las evaluaciones de toxicología revelaron que la administración del extracto no produjo muertes en los animales de experimentación, ni cambios orgánicos o de comportamiento. El compuesto mayoritario identificado en el extracto activo fue escopoletina.

Phytochemical and biological study of vangueria edulis cultivated in Egypt

Fractionation and purification of the alcoholic extract of the leaves and stem -bark of Vangueria edulis cultivated in Egypt afforded thirteen compounds identified as: p-sitosterol acetate [1], stigmasterol [2], palmitic acid [3], scopoletin [4], p-coumaric acid [5], protocatechuic acid [6], esculetin [7], ethyl I-O-glucosyi-4-O-[E] caffeoyl quinate [8], kaempferol 3-O-rhamnoside, 7-O-rutinoside [9], p-sitosterol [10], ceryl alcohol [11], vanillic acid [12] and p-sitosterol-3-Q-beta-D-glucopyranoside [13]. Identification of these compounds has been established by physical, chemical and spectral data as well as comparison with authentic samples. Compounds 8 and 9 are firstly reported from a natural source while the rest of the compounds have been isolated for the first time from the genus Vangueria. The LD[50] antibacterial antifungal anti-inflammatory, antipyretic and the effect on CNS were studied

Chemical constituents from stems of Dysoxylum laxiracemosum / 中国中药杂志

Twelve compounds were separated from stems of Dysoxylum laxiracemosum and their structures were identified by spectrum analysis as shoreic acid (1), cabraleahydroxylactone (2), cabralealactone (3), cinchonain (5), catechin (6), scopoletin (7), vanillic acid (8), p-hydroxybenzoic acid (9), docosanol (10), beta-sitosterol (11), daucosterol (12). Of them, compounds 1-6,8-12 were separated from this plant for the first time, and compounds 4-6 were reported from this plant genus for the first time.

Chemical constituents of Eupatorium lindleyanum / 中国中药杂志

To study chemical constituents of Eupatorium lindleyanum. Ethyl acetate extractive fractions were separated with silica gel and Sephadex LH-20 by column chromatography, and their structures were identified on the basis of spectroscopic analysis and chemical evidence. Sixteen compounds were separated and identified as scopoletin (1), 6, 7-dimethylesculetin (2), nepetin (3), eupatrin (4), luteolin (5), isoquerecitrin (6), jaceosidin (7), quceritin (8), kaempferol (9), rutin (10), cirsiliol (11), taraxasterylacetate (12), pseudotaraxasteryl acetate (13), pseudotaraxasterol (14), butanoic acid (15) and n-hexadecanoic acid (16). Of them, compounds 1-6 and 11, 13 and 15 were separated from this plant for the first time.

Main chemical components for clearing heat and relieving sore throat in different germplasm of Canarium album / 中国中药杂志

<p><b>OBJECTIVE</b>To screen the representing components in different germplasms of Canarium album and determine the content by HPLC for laying the foundation for the quality specification.</p><p><b>METHOD</b>Gallic acid and scopoletin and scoparone were chosen as the main chemical components for clearing heat and relieving sore throat by reviewing literature and qualitative experiment. Gallic acid was determined according the method of Galla chinensis in Chinese pharmacopoeia. Scopoletin and scoparone were determined by HPLC on C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase was acetonitrile-water-acetic acid (16:84:0.5) with the detection wavelength at 345 nm and column temperature at 35 degrees C , the flow rate was 1.0 mL x min(-1).</p><p><b>RESULT</b>The resolution is good for gallic acid and scopoletin and scoparone. The standard curve in detection range showed good linear relation, the average recoveries were 100.22%, 101.22% and 100.42%, respectively, RSD were all less than 2.0%. The content of gallic acid in the large fruit C. album from Guangdong was higher than that in the small fruit C. album, while the result was contrary in samples from Sichuan. The content of total coumarins in the small fruit C. album was generally higher than that in the large fruit C. album, which from Sichuan was higher than those from Guangdong.</p><p><b>CONCLUSION</b>The efficacies of clearing heat and relieving sore throat in C. album were formed by synergistic effect of gallic acid and scopoletin and scoparone. The small fruit C. album was better than the large fruit C. album as medicinal materials, the results provide reliable basis for quality control in medicinal materials and its preparation.</p>

Study on chemical constituents in roots and rhizomes of Ligularia duciforms II / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents in roots and rhizomes of Ligularia duciforms.</p><p><b>METHOD</b>90% ethanol extract was isolated and purified by silica gel and sephadex LH-20 column chromatography, the structures of compounds were identified by physicochemical properties and spectral analysis.</p><p><b>RESULT</b>Nine compounds were isolated and identified as lupeol (1), isoscopoletin (2), isoline (3), duciformine (4), (2S, 3S, 4R)-sphinganine-(2'R)-delta5',6' (E)-2'-hydroxytetracosanoylamino (5), tetracosanoic acid (6), tetracosanoicacid glyceride (7), (E)-docosyl ferulate (8), (E)-docosyl caffeate (9).</p><p><b>CONCLUSION</b>Compounds 1, 2, 5-7 were isolated for the first time from this plant, and the compound 2, 5-7 were isolated firstly from the genus Ligularia.</p>

Studies on chemical constituents of Saussurea laniceps / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of Saussurea laniceps.</p><p><b>METHOD</b>The ethanol extract of S. laniceps was separated by means of silica gel chromatography. The compounds isolated from the plant were identified by their spectral evidence.</p><p><b>RESULT</b>Fifteen compounds were isolated and identified as beta-stiosterol (1), umbelliferone (2), 4-hydroxyacetophenone (3), scopoletin (4), isoscopoletin (5), xuelianlactone (6), methyl 3-(2', 4'-dihydroxyphenyl) propanoate (7), apigenin (8), neoechinulin A (9), daucosterol (10), scopolin (11), xuelianlactone 8-O-beta-D-glcuoside (12), apigenin 7-glcuoside (13), apigenin 7-lutinoside (14) and syringin (15).</p><p><b>CONCLUSION</b>Compounds 5-15 were isolated from S. laniceps, and among them, 7 and 9 were isolated from genus Saussurea for the first time.</p>

Studies on absorption kinetics of scopoletin in rat stomachs and intestines / 中国中药杂志

<p><b>OBJECTIVE</b>To study absorption kinetics of scopoletin in rat stomachs and intestines.</p><p><b>METHOD</b>Rats was cannulated for in situ recirculation. UV and HPLC methods were used to determine the concentrations of phenolsulfonphthalein and scopoletin, respectively.</p><p><b>RESULT</b>The absorption rates in rat stomachs at 2 h after administration was 76.31%; The absorption rates at colon, duodenum, ileum and jejunum were 46.25%, 40.54%, 38.21%, 32.77%, respectively. The absorption rate constant (Ka) at concentrations of 10.0144, 20.0288-40.0576 mg x L(-1) in intestine were 0.6434, 0.6137, 0.5970 h(-1), respectively. The Ka of scopoletin at pH of 6.0, 6.8 and 7.4 in intestine were 0.6217, 0.6033, 0.6137 h(-1), respectively.</p><p><b>CONCLUSION</b>The concentrations and pH values of scopoletin solution had no distinctive effect on the absorption kinetics. The absorption of scopoletin was a first-order process with passive diffusion mechanism. Scopoletin was well absorbed at stomachs and intestines in rats. Colon was the best absorption site of scopoletin, which suggest that a sustained-release preparation should be suitable for this compound.</p>

Studies on chemical constituents of Ranunculus japonicus / 中国中药杂志

<p><b>OBJECTIVE</b>To isolate and determine the chemical constituents of Ranunculus japonicus in Liupan Mountain, Ningxia province, China.</p><p><b>METHOD</b>The herb was extracted with ethanol by ultrasonic bath. The extractives were divided to petroleum ether, ethyl acetate, n-butanol parts. The first two parts were separated and purified with silica gel and Sephadex LH -20 column chromatography. The structures of the separated compounds were idnetefied by physical and chemical properties and spectral analysis.</p><p><b>RESULT</b>Nine compounds were isolated and identified as follows: scoparone (1), tricin (2), protocatechuic acid (3), luteolin (4), anemonin (5), scopoletin (6), 5-hydroxy-6, 7-dimethoxyflavone (7), ternatolide (8), 5-hydroxy-7, 8-dimethoxy-flavone (9).</p><p><b>CONCLUSION</b>Compounds 1-9 were isolated from Ranunculus japonicus for the first time.</p>