Chemical constituents and in vivo preliminary evaluation of the toxicological activity of Ouratea spectabilis (OCHNACEAE) and Clitoria guianensis (Fabaceae) leaves

Clitoria guianensis and Ouratea spectabilis, found in the Brazilian Cerrado, are used in folk medicine, despite the few chemical and biological studies reported in the literature. The present study aims to investigate the toxicity and effect of extracts from both species on the microcrustacean Artemia salina, and to determine the chemical composition of the hexane extract of O. spectabilis leaves and the EtOAc fraction of C. guianensis leaves. Kaempferitrin, a flavonoid isolated from of the EtOAc fraction of C. guianensis leaves, was identified by chemical analysis. Analysis of the hexane extract of O. spectabilis leaves using gas chromatography-mass spectrometry (GC-MS) suggested the presence of twenty-five known substances. The Hex, EtOAc, and EtOH crude extracts of C. guianensis leaves exhibited high and moderate toxicity against Artemia salina, with median lethal dose values (LD50) of 43.7, 25.4, and 233.4 mg.L−1, respectively. The acetone extract of O. spectabilis leaves showed moderate toxicity against Artemia salina with an LD50 value of 115.13 mg.L−1.

Decrease in Dengue virus-2 infection and reduction of cytokine/chemokine production by Uncaria guianensis in human hepatocyte cell line Huh-7

ABSTRACT BACKGROUND Dengue fever may present hemorrhages and cavitary effusions as result of exacerbated immune responses. We investigated hydro-alcoholic extracts from leaves (UGL) and bark (UGB) of the medicinal species Uncaria guinanensis with respect to antiviral effects in Dengue virus (DENV) infection and in immunological parameters associated with in vivo physiopathological features. METHODS Chemical profiles from UGB or UGL were compared in thin layer chromatography and 1H nuclear magnetic resonance using flavonoid compounds and a pentacyclic oxindole alkaloid-enriched fraction as references. DENV-2-infected hepatocytes (Huh-7) were treated with extracts. Cell viability, DENV antigens and immunological factors were detected by enzyme-linked immunosorbent assay (ELISA) or flow cytometry. FINDINGS The UGL mainly differed from UGB by selectively containing the flavonoid kaempferitrin. UGB and UGL improved hepatocyte viability. Both extracts reduced intracellular viral antigen and inhibited the secretion of viral non-structural protein (NS1), which is indicative of viral replication. Reduction in secretion of macrophage migration inhibitory factor was achieved by UGB, of interleukin-6 by UGL, and of interleukin-8 by both UGB and UGL. MAIN CONCLUSIONS The U. guianensis extracts presented, antiviral and immunomodulatory effects for DENV and possibly a hepatocyte-protective activity. Further studies may be performed to consider these products as potential candidates for the development of an herbal product for the future treatment of dengue.

Inspection of Illegal Use of Woodwardia unigemmata in Kanggan Granules by HPLC-MS/MS / 中国药学杂志

OBJECTIVE: To establish an HPLC-MS/MS method for the inspection of illegal use of Woodwardia unigemmata, an adulterant of Dryopteris crassirhizoma, in Kanggan granules. METHODS: The content of kaempferitrin in Woodwardia unigemmata is significantly higher than that in Dryopteris crassirhizoma. Therefore, the content of kaempferitrin can reflect the material usage(Woodwardia unigemmata or Dryopteris crassirhizoma) in Kanggan granules. C18 column was applied with a mobile phase consisting of 0.02% formic acid acetonitrile(V/V)-0.02% formic acid H2O(V/V) in gradient elution mode. The mass spectral analysis was performed in a positive electrospray ionization mode, and scanned by a multiple reaction monitoring mode. The precursor/product ion pair used for quantification was m/z 579.2→287.1. RESULTS: A good linear relationship was obtained in the range of 272.46-5 449.25 pg for kaempferitrin, with the linear regression equation of Y=139.217 2X-2 901.93(r=0.999 6); the average recovery was 94.97%(RSD=0.85%, n=6). Among 12 batches of Kanggan granules, 7 batches from 2 enterprises showed abnormally high contents of kaempferitrin, which indicated illegal use of Woodwardia unigemmata. CONCLUSION: The method is simple, accurate, and reliable, which can be applied to detect the use of Woodwardia unigemmata in Kanggan granules.

UPLC fingerprint of Viciae Amoenae Herba / 中草药

Objective: To establish the ultra performance liquid chromatography (UPLC) fingerprint of Vicia amoena var. angusta and compare the other five different origin varieties of Viciae Amoenae Herba in order to evaluate the quality of the preparation. Methods: Eclipse Plus C18 RRHD column (50 mm×2.1 mm, 1.8 μm) were used with acetonitrile (B) and 0.1% formic acid aqueous solution (A) in gradient elution mode at a flow rate of 0.2 mL/min. The detection wavelength was set at 340 nm and the column was 30℃. The fingerprints for V. amoena var. angusta from 10 different areas were set up, the similarity assay for V. amoena var. angusta from ten different areas was carried out to evaluate their quality by Chinese Materia Medica (CMM) Fingerprint Similarity Evaluation System (2004A edition), and the peaks were identified by comparing the reference substance with LC-MS/MS; Whilie the other origins of five kinds of varieties of Viciae Amoenae Herba were compared and identified in the same condition. Results: UPLC fingerprints for V. amoena var. angusta were established, six chromatographic peaks among 15 common peaks were identified by UPLC-ESI-Q-TOF-MS and be intended to be chlorogenic acid, myricitrin, hyperoside, isoquercitrin, kaempferit-rin, and quercitrin. UPLC fingerprint mutual pattern of other five kinds of origins varieties of Viciae Amoenae Herba and V. amoena var. angusta had obvious differences. Conclusion: This method is simple, rapid, stable, and reproducb1e, and could be used for the full quality evaluation of Viciae Amoenae Herba.

Determination of kaempferitrin in Celastri Orbiculati Fructus by HPLC / 中草药

Objective: To establish an HPLC method for the determination of kaempferitrin in Celastri Orbiculati Fructus and to compare the quality of samples from different habitats in northeast China. Methods: The extracts were obtained by methanol ultrasonic method and kaempferitrin was determinated by HPLC. Chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C18 column (250 mm × 4.6 mm, 5 μm) at 30℃ and the mobile phase was a mixture of methanol- 0.01 mol/L solution of potassium dihydrogenphosphate -glacial acetic acid (40:60:15). The flow rate was 1.0 mL/min and the detection wavelength was 254 nm. Results: The linearity range was 0.092 7-0.926 7 μg (r = 0.999 8), average recovery was 100.8%, and RSD = 1.1% (n = 6). Conclusion: This method is validated in terms of selectivity, linearity, precision, and accuracy. It can be used as the effective method for quality control. The determination results show that the longer the fruit sits followed by picking, the less contents it has.

Controle de qualidade de drogas vegetais a base de Bauhinia forficata Link (Fabaceae) / Quality control of drugs with Bauhinia forficata Link (Fabaceae)

Este trabalho teve como objetivo utilizar métodos de controle de qualidade para algumas drogas vegetais a base de Bauhinia forficata comercializadas nos municípios de Itajaí e Balneário Camboriú, utilizando como marcador químico o flavonóide kaempferitrina. As metodologias empregadas foram cromatografia em camada delgada (CCD) e cromatografia líquida de alta eficiência (CLAE), além da análise microscópica, análise de rótulos e características organolépticas e a presença de material estranho. Todas as análises foram comparadas com os resultados demonstrados pela amostra autêntica, devidamente identificada, de B. forficata (A). Foram analisadas 06 amostras a base de Bauhinia identificadas através de siglas, sendo a B. forficata autêntica a amostra A e seis comerciais denominadas B-G. Todas as amostras comerciais foram reprovadas quanto à presença de material estranho, ficando acima de 2 por cento. Na análise microscópica observou-se que apenas a amostra E e G apresentam-se iguais à B. forficata. A análise cromatográfica revelou a presença de kaempferitrina somente nas amostras B, C, E e G.

The present study aimed to use methods to perform the quality control of some vegetal drugs based on Bauhinia forficata from Itajaí and Balneário Camboriú, using the flavonoid kaempferitrin as chemical marker. The methods used were thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), besides microscopic analysis, label analysis and organoleptic characteristics and strange material presence. All the analyses were compared with the authentic sample of B. forficata (A) results. This study used six commercial samples which were denoted as B-G. In the strange material analyses, all the samples were disapproved because they were above 2 percent. Microscopic analyses revealed that only B and G samples were identical to B. forficata (A). The chromatographic results suggested that only the samples B, C, E and G presented kaempferitrin.