[Study and comparison on HPLC fingerprints of flavonoids of frequently used Chinese materia medica in citrus].

OBJECTIVE: To establish the HPLC fingerprint of flavonoids of the six clinical frequently used Chinese materia medica for regulating Qi flow,such as Citri grandis, C. grands, Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, Aurantii Fructus, and Aurantii Fructus Immaturus from Citrus, and analysis differences in the fingerprints to provide scientific basis for profile-effect research and clinical reasonable use. METHOD: HPLC was performed on a C18 column with methanol-water (with acetic acid), to establish HPLC fingerprints of the six kinds of medicinal herbs on the same chromatograph condition. RESULT: The six frequently used Chinese materia medica were divided into naringin type and hesperidin type according to the method of phytochemotaxonomy. Based on the retention time of chromatograph peaks, C. grandis and C. grands had fifteen common peaks; Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, Aurantii Fructus and Aurantii Fructus Immaturus had ten common peaks. All herbs had five common peaks. Compared with mutual model, the holistic similarity of chromatograms of C. grandis and C. grands was in the range of 0.9285 - 0.9962. The degree of similarity was high. For Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride and Aurantii Fructus Immaturus, it was in the range of 0.9221 - 0.9973 and high. But the similarity of Aurantii Fructus was only in 0.4547 - 0.7733 with the mutual model. CONCLUSION: The established fingerprints of flavonoids of the six common traditional Chinese medicines can be used to compare the differences intuitively. Meanwhile, the peak height and peak areas of characteristic peaks are different remarkably, but whether it is connected with the different function of regulating Qi flow of the six medical materials in clinical use, is still needed to be studied.

[AAPH scavenging activities of 22 flavonoids and phenolic acids and 9 extracts of Chinese materia medica].

OBJECTIVE: To investigate the AAPH scavenging activities of 22 flavonoids and phenolic acids and 9 extracts of Chinese materia medica. METHOD: The antioxidant activities of the samples were evaluated by an oxygen radical absorbance capacity method (ORAC), at the same time, the total contents of flavonoids and phenolic the 9 herb extracts were analyzed by Folin-Ciocalteu method, and the active components were qualitatively and quantitatively analyzed by an HPLC method. RESULT: It was found that the tea extract showed the strongest AAPH activity with the ORAC value of 4786.40 micromol x g(-1) whereas safflower demonstrated the weakest activity with the ORAC value of 784.04 micromol x g(-1). As for compounds, quercetin had the strongest AAPH activity with the ORAC value of 12.90 while ( - )-EGC had the weakest activity with the ORAC value of 2.47. A quantitative relationship was obtained to describe the AAPH scavenging activity of the herb extracts: Y = 1844.8 lnX-3577.5, r = 0.8675, where Y stands for the ORAC vaule, and X stands for the concentration of total phenolic acids. CONCLUSION: Flavonoids and phenolic acids are the AAPH scavenging active ingredients in the Chinese herb extracts. It's a good way to study the antioxidant activity of Chinese herb extract and its chemical composition by combing ORAC method and HPLC method.

Antiproliferative potential and phenolic compounds of infusions and essential oil of chamomile cultivated with homeopathy.

ETHNOPHARMACOLOGICAL RELEVANCE: Chamomilla recutita (Asteraceae) is used worldwide as a soothing, anti-inflammatory and aromatherapy. In Brazil, it is one of the most cultivated medicinal species. However, the cultivation form may alter the production of compounds in the secondary metabolism and compromise the therapeutic purpose of this species. AIM OF THE STUDY: Evaluation of antiproliferative and genotoxic effects of infusions and essential oil of chamomile, cultivated with homeopathy, on the cell cycle of Allium cepa, as well as the determination of the phenolic compounds present in the infusions of the chamomile inflorescences. MATERIAL AND METHODS: For the Allium cepa test, two concentrations of 10 and 40 g L-1 of inflorescences of chamomile were used for the preparation of the infusions and essential oil diluted to 0.10%, referring to the six treatments obtained in field cultivation, in which were carried out the applications of homeopathy from the emergence to the harvest of the plants. Distilled water and ethanol were used as negative control and glyphosate 2% as a positive control. The determination of phenolic compounds present in the infusions was carried by liquid chromatography in a UHPLC apparatus. RESULTS: Chamomile infusions at concentrations of 10 and 40 g L-1 of inflorescence reduced mitotic index and emphasized antiproliferative activity on the cell cycle of Allium cepa. However, the treatments related to essential oil diluted to 0.10% showed a response variation dependent on the dynamization used, as well as for apigenin concentration. CONCLUSIONS: Therefore, cultivation with homeopathy does not induce a genotoxic effect in the use of infusions and essential oil of chamomile and it emphasize antiproliferative activity on the cell cycle of Allium cepa, favoring the sustainable cultivation and the safe use of this medicinal species when cultivated with homeopathy.

[Simultaneous determination of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill by RP-HPLC].

OBJECTIVE: A new method for the simultaneous quantitative determination of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill using reversed phase high performance liquid chromatographic method had been developed. METHOD: The optimum chromatographic conditions were as follows: Agilent Zorbax SB - C18 column (4.6 mm 250 mm, 5 m), acetonitrile-H2O (containing 6 mmol L(-1) KH2PO4, pH 4.6) gradient elution; as a detection wavelength of 343 nm. RESULT: The calibration curves of geniposide, baicalin and berberine were linear at the ranges of 4.50-110.00, 5.00-153.00, 6.40-191.00 mg L(-1), respectively. The limits of detection of the method were 0.77 ng for geniposide, 1.53 ng for baicalin and 1.43 ng for berberine hydrochloride. The recoveries of the method were 104.44% (RSD 1.79% ) for geniposide, 96.98% (RSD 1.76%) for baicalin, 101.08% (RSD 3.1%) for berberine hydrochloride. CONCLUSION: This method had been successfully applied to determine the content of geniposide, baicalin and berberine hydrochloride in Angong Niuhuang pill.

[Antioxidant activity of natural and cultured Cordyceps sp].

OBJECTIVE: Investigating the antioxidant activities of water and ethanol extracts of natural Cordyceps sinensis and Cordyceps militaris and their fermentation preparations. METHOD: The samples were tested through 6 assays: inhibition ability of linoleic acid oxidation; scavenging activity of DPPH, hydrogen peroxide, hydroxyl radical and superoxide anion; and metal chelating activity. RESULT: Samples showed different antioxidant ability, and there was not an extract that exhibited high activity in all assays; however, water extract of natural C. militaris could be regarded as the most powerful antioxidant among 8 samples. It had high activity in inhibition of linoleic acid oxidation, chelating metal ions, and scavenging DPPH and hydroxyl radical. The research also indicated that the contents of phenolic compounds in water and ethanol extracts of natural and cultured Cordyceps sp. had huge difference. CONCLUSION: Natural Cordyceps sp. and its fermentation preparations could be used as potential natural antioxidants. The fermented process affected the antioxidant ability of cultured Cordyceps sp., and the antioxidant activity of both natural and cultured Cordyceps sp. did not significantly related with the quantity of phenolics.

[Study on HPLC fingerprint and quality control of Propolis].

OBJECTIVE: To study on HPLC fingerprint of Propolis and to control it's quality. METHOD: The chromatographic fingerprints of seven samples from different producing areas were determined by RP-HPLC. RESULT: The chromatograms of Propolis from different producing areas were very similar. CONCLUSION: The quality of Propolis can be controlled by determination the HPLC fingerprint.

[Compatible leaching effect of Concha Ostreae and some commonly-used Chinese medicinal materials].

OBJECTIVE: Based on a study of Concha Ostreae, to provide some reference for the rational approach to leaching mineral drugs. METHOD: Taking the change rates of compatible leaching and value of A as targets, the compatible leaching effect of Concha Ostreae with Radix et Rhizoma Rhei Praeparata, Radix Scutellariae, Pollen Typhae and Fructus Aurantii lmmaturus was studied. RESULT: If Concha Ostreae and the above cited medicinal herbs are decocted together, the yield of extract may be reduced to a certain extent, but the target composition of Radix et Rhizoma Rhei Praeparata and Fructus Aurantii lmmaturus remains intact, only the content of Radix Scutellariae baicalin may be influenced. CONCLUSION: The relationship of technology with preparation must be taken into consideration when making choice of the proper way to leach Concha Ostreae.

[Studies on extraction factors of buxueyangyan mixture].

OBJECTIVE: To study the optimum extraction factors of Buxueyangyan Mixture. METHOD: The orthogonal design was used to select the optimum extraction factors of Cervi Pantotrichum etc. and water extraction rate was used to evaluate the factors. The orthogonal desingn was used to study three factors including crude particle size, extraction time and amount of water. Lcariine content was used as analytical parameter. RESULT: The optimum extraction factors for Cervi Pantotrichum etc. were: 3 a volume ofwith 10 times of water used for three extraction 2.5 h each time. The optimum extraction factors for others were: 1 cm or smaller the crude particle size, boiling 2 times: 2.5, 1.5 h respectively the amount of water used was 10. 8 times of the drug mixture respectively. CONCLUSION: According the optimum extraction factors, the active substance can be extracted efficiently.

[Preparation of huichun zhibao oral liquid (HZOL)].

The formulation, preparing method, quality standards, pharmacodynamic experinents and toxicological experiments of HZOL are described. The content of icariin, which is and effective component of epimedium herb as well as one of the main ingredients in HZOL, was successfully determined through HPLC, with an average recovery of 98.98% and RSD = 1.53%.

Analytical characterisation of homoeopathic mother tinctures.

Quality of homoeopathic mother tinctures is assured by the definition of the starting material, the manufacturing process and the analytical characteristics described in the monograph. Traditionally analytical characterisation of the mother tincture comprises appearance, odour, identity, density and dry residue. According to annex I of directive 2001/83/EC an assay is only performed in case of a health hazard due to toxic compounds. The concept of marker substances as usually used in phytotherapy cannot be transferred to mother tinctures without research effort. For example the marker substances echinacoside, apigenin-7-glucoside and rosmarinic acid found in dried underground parts of Echinacea pallida Nutt., dried flower heads of Matricaria recutita L. and dried herb of Pulmonaria officinalis L. cannot be found in homoeopathic mother tinctures prepared from fresh material thereof.

Inhibition of acetylcholinesterase activity by hidroalcoholic extractand their fractions of Bouvardia ternifolia (Cav. ) Shcltdl (Rubiaceae) / Inhibición de la actividad de acetilcolinesterasa por el extracto hidroalcohólico y sus fracciones de Bouvardia ternifolia (Cav. ) Shcltdl (Rubiaceae)

The hydroalcoholic extract (BtHA) and its fractions of Bouvardia ternifolia were evaluated as inhibitors of the activity of the acetylcholinesterase enzyme utilizing the in vitro method (Ellman). BtHA inhibited the acetylcholinesterase enzyme competitively (IC50 = 0.6 ug/ml); the ethyl acetate fraction (BtF-AcOEt) caused mixed-type inhibition (IC50 = 0.96 ug/ml). A fraction insoluble on methanol (Bt-Faq-1) showed a mixed-type inhibition (CI50 = 0.96 ug/ml). Finally, the methanol-soluble fraction (Bt-Faq-2), presented complex, mixed-type inhibition that corresponds to the C5 system ( alpha= 0.740 and beta = 0.842). Rutin, quercetin, kaempferol and ursolic acid were detected by HPLC and the concentration of these compounds was different in each fraction.

El extracto hidroalcohólico (BtHA) y fracciones provenientes de Bouvardia ternifolia fueron evaluadas como inhibidores de la enzima acetilcolinesterasa utilizando el método enzimático propuesto por Ellman. BtHA inhibe a la enzima de manera competitiva (IC50 = 0.6 ug/ml); la fracción de acetato de etilo (BtF-AcOEt) provoca una inhibición mixta (IC50 = 0.96 ug/ml). La fracción insoluble en metanol (Bt-Faq-1) mostró una inhibición tipo mixta (CI50 = 0.96 ug/ml). Finalmente la fracción soluble en metanol, Bt-Faq-2, inhibe a la enzima presentando una inhibición mixta que corresponde a un sistema C5 ( alfa= 0.740 and beta= 0.842). Mediante HPLC se detectó rutina, quercetina, canferol y ácido ursólico la concentración de estos compuestos fue diferente en cada fracción.

Quality criteria of homoeopathic mother tinctures: considerations regarding suitable tests for homoeopathic monographs.

Quantitative determination of markers may improve quality control of herbal homoeopathic mother tinctures. Since the activity of homoeopathic medicinal products does not depend on the specific content of such markers, appropriate substances can be selected according to analytical aspects only. We tested carbohydrates, amino acids, total polyphenols and flavonols in different mother tinctures. The results obtained with the latter two groups of substances imply that corresponding tests might be useful for inclusion into pharmacopoeial monographs.