Site-specific accumulation and dynamic change of flavonoids in Apocyni Veneti Folium.

Site-specific accumulation of flavonoids in Apocyni Veneti Folium was determined by laser scanning confocal microscope (LSCM) and the localization of catechins also was observed via vanillin-HCl staining under the conventional optical microscope. The contents of five flavonoids in Apocyni Veneti Folium from different harvest times and growth parts were measured using HPLC method. LSCM observation showed that flavonoids are accumulated in cuticle of epidermal cells and vessel walls, especially in protoplasts and nucleolus of the collenchyma cells and the epidermal cells. Catechins are localized in the palisade parenchyma cells and vessel walls, particularly in the laticifers found in the phloem. On the basis of the difference of the maximal emission wavelength between quercetin and kaempferol derivatives which have fluorescence behavior by appropriate treatment, kaempferol and its derivatives are localized exclusively in the cuticle. Results showed that the content of astragalin in Apocyni Veneti Folium from different parts revealed the decreasing trend, while hyperin and isoquercitrin were higher in June and July analyzed by HPLC. In summary, the site-specific accumulation of flavonoids in Apocyni Veneti Folium can be determined by LSCM and vanillin-HCl staining. The contents of flavonoids in Apocyni Veneti Folium are correlated with harvest times and growth parts.

Catalpol suppressed proliferation, growth and invasion of CT26 colon cancer by inhibiting inflammation and tumor angiogenesis.

Tumor angiogenesis and inflammation, which play important roles in mediating tumor proliferation and growth, should be inhibited to effectively regulate tumor progression. Catalpol, a main active ingredient extracted from Rehmannia glutinosa, has various pharmacological actions including anti-apoptotic, anti-inflammatory, hypoglycemic and anti-cancer properties. However, the pharmacological effect of catalpol on colon cancer remains largely unknown. The aim of this study was to investigate the effects of catalpol on the proliferation, growth, invasion, tumor angiogenesis and inflammation of CT26 colon cancer in vitro and in vivo. Catalpol inhibited the proliferation and growth of CT26 cells in concentration- and dose-dependent manners in vitro and in vivo, respectively. Catalpol suppressed the invasion of CT26 cells in vitro. Tumor cell-induced vascularization of endothelial cells and rat aortic ring angiogenesis were impaired by catalpol. Catalpol reduced the secretions of several angiogenic markers in the culture supernatant of CT26 cells. Immunohistochemical assay showed that catalpol inhibited the expressions of angiogenic markers vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2), hypoxia inducible factor-1α (HIF-1α) and basic fibroblast growth factor (bFGF) in colon cancer tissues. Moreover, catalpol inhibited the expressions of inflammatory factors interleukin-1ß (IL-1ß), IL-6, IL-8, cyclooxygenase (COX-2) and inducible nitric oxide synthase (iNOS). Taken together, catalpol suppressed the growth and invasion of CT26 colon cancer cells mainly by inhibiting inflammation and tumor angiogenesis, as a promising candidate compound for treating colon cancer.

[Study on the HPCE fingerprint of Rehmannia glutinosa].

OBJECTIVE: To establish the analytical method for the fingerprint of Rehmannia glutinosa by HPCE and compare the fingerprints of Rehmannia glutinosa and its processed products. METHODS: Based on the mode of high performance capillary electrophoresis, 60 mmol/L sodium borate was used as buffer solution (5% MeOH, pH 9.5). The separation voltage was 20 kV and the detection wavelength was set at 210 nm. Catalpol was used as a reference standard, the chromatographic fingerprint were determined. The data were analyzed by fuzzy cluster and fingerprint similarity evaluation software was used to compare the similarity of samples. RESULTS: HPCE fingerprints with 7 common peaks of Rehmannia glutinosa were established preliminarily. It was discovered that a small number of samples differed from others. Regarding to the fingerprints of Rehmannia glutinosa and its processed products, there were obvious differences in the relative areas of common peaks. CONCLUSION: The method is reliable, accurate and can be used for quality control of Rehmannia glutinosa.

[Study on HPCE fingerprint of Eucommiae Cortex].

OBJECTIVE: To establish the analytical method for the fingerprint of Eucommiae Cortex by HPCE and compare the fingerprints of Eucommiae Cortex and its processed products. METHODS: Based on the mode of high performance capillary electrophoresis, 60 mmol/L sodium borate-20 mmol/L sodium dihydrogen phosphate-10% methanol (pH 10.0) was used as buffer solution. The separation voltage was 20 kV and the detection wavelength was set at 210 nm. Pinoresinol diglucoside was used as a reference standard, the chromatographic fingerprint were determined. The data were analyzed by fuzzy cluster and fingerprint similarity evaluation softwarewas used to compare the similarity of samples. RESULTS: HPCE fingerprints with 10 common peaks of Eucommiae Cortex were established preliminarily. It was discovered that a small number of samples differed from others. Regarding to the fingerprints of Eucommiae Cortex and its processed products, there were obvious differences in the relative areas of common peaks. CONCLUSION: The method is reliable, accurate and can be used for quality control of Eucommiae Cortex.

[Comparative analysis of volatile components of medicinal materials from Perilla frutescens].

OBJECTIVE: To analyze the differences of volatile components of medicinal materials from Perilla frutescens (Perillae Folium; Perillae Caulis; Perillae Fructus). METHODS: The volatile components were identified by HSGC/MS. The relative percentage content of the components were determined by peak area normalization method and the content change of common components were determined. RESULTS: 47 compounds were separated and identified from Perillae Folium, 39 compounds were identified from Perillae Caulis, 46 compounds were identified from Perillae Fructus. There were 17 common components of the three groups. For example, beta-Caryophyllene, Perillaketone, Caryophyllene, Limonene, Linalool, 1-Octen-3-ol, alpha-Bergapten and Apiol. CONCLUSION: It provides basis scientific explanation to the differences of traditional efficacy of medicinal materials from Perilla frutescens.

[Preliminary analysis of quality of Antrodia camphorata powder].

OBJECTIVE: To proceed a preliminary analysis of the quality of Antrodia camphorata powder. METHODS: The contents of water-soluble extract were detected according to the standards stated in Chinese Pharmacopoeia. UV-VIS was used to analyze total polysaccharide and triterpenoid. HPLC method was applied for the analysis of adenosine in Antrodia. camphorata. Besides, volatile compounds were analyzed by HSGC-MS. RESULTS: The contents of water-soluble extract (37.26% - 40.98%), total polysaccharide (5.45% - 8.08%), total triterpenoid (2.44% - 2.87%) and adenosine (0.0470% - 0.0604%) were obtained respectively. 49 volatile compounds were identified in Antrodia camphorata powder. CONCLUSION: The established method can be used for the quality control of Antrodia camphorata powder.

[Study on HPCE-DAD fingerprint of Salviae Miltiorrhizae Radix et Rhizoma].

OBJECTIVE: To establish the analytical method for the fingerprint of Salviae Miltiorrhizae Radix et Rhizoma by HPCE-DAD and estimate its quality. METHODS: Salviae Miltiorrhizae Radix et Rhizoma were analyzed and the chromatographic fingerprint were determined by HPCE-DAD. The data were analysed by fuzzy cluster and fingerprint similarity evaluation software was used to compare the similarity of samples. RESULTS: HPCE-DAD fingerprint of 10 main common peaks was established preliminarily. It was discovered that a small number of samples were different from the others. CONCLUSION: The method is reliable, accurate and can be used for the quality control of Salviae Miltiorrhizae Radix et Rhizoma

[Simultaneous determination of six flavonoids in Hyperici Japonici Herba by HPCE-DAD].

OBJECTIVE: To establish a high performance capillary electrophoresis method with diode array detection (HPCE-DAD) for simultaneous determination of rutin, isoquercitrin, hyperoside, quercitrin, kaempferol and quercetin in Hyperici Japonici Herba. METHOD: Based on the mode of capillary zone electrophoresis, 40 mmol x L(-1) borax was used as buffer solution (pH 8.62), uncoated fused silica capillary (56 cm x 64.5 cm x 75 microm) was used, separation voltage was 25 kV, detection wavelength was at 206 nm, column temperature was maintained at 25 degrees C, and sample was injected at 50 mbar, 8 s. RESULT: Six flavonoids showed good linearity (r > 0.9953) in the range of the tested concentration, the average recoveries of the method were between 98.8%-102.9%. CONCLUSION: The method is simple, accurate and reproducible, and can be used for quality control of Hyperici Japonici Herba.

[Location and relative quantity of flavonoids in the leaf of Apocynum venetum].

In this study, laser scanning confocal microscopy (LSCM) was used to determine the location and relative quantity of flavonoids in the leaves of Apocynum venetum L. from the top, middle and basal parts of the branch. The leaves of the plants of one, two and three years old, separately, were collected in July. ANOVA and LSD test were employed in the statistical analysis. The results indicated that flavonoids located mainly in xylem conduit of vein, collenchyma, epidermic cells and cuticle. The data of flavonoids contents under statistical analysis showed that difference existed in the leaves of different parts and different ages. This study provided the reliable scientific material about the analysis of the ecological and the exploitation of the leaves of Apocynum venetom L.

[Study on water-soluble iron, heavy metals and harmful elements of Magnetitum].

OBJECTIVE: To measure the contents of the water-soluble iron, five heavy metals and harmful elements in Magnetiturn and provide a basis for the quality control and safety evaluation of Magnetitum. METHOD: Iron (Fe), lead (Pb), cadmium (Cd) and copper (Cu) were determined by atomic absorption spectrometry (AAS); arsenic (As) and mercury (Hg) were determined by atomic fluorescence spectrometry (AFS). RESULT: The mean content of element iron is 764.30 mg x kg(-1). The contents of five water-soluble heavy metals and harmful elements in Magnetitum were within the safety range. The recovery of the standard addition was in the range of 93.7% - 110.6%, and the RSD was less than 5.0%. CONCLUSION: Analyzing the water-soluble iron, heavy metals and harmful elements in Magnetitum is effective to the quality control and the safety evaluation of magnetitum.

[Study on the fingerprints of magnetitum by FTIR].

Thirteen different magnetitum samples were analyzed by FTIR, and the FTIR fingerprint was set up with them. Processed samples and the crudes were compared with the fingerprint. It was found that all the similarities of the samples are more than 0.97. The similarities and the correlation coefficients of the processed samples are decreased. The FTIR fingerprint could be used for evaluating the quality of magnetitum for commodities.

[X-ray diffraction Fourier fingerprint of mineral Chinese medicine Chloriti Lapis].

The technology of powder X-ray diffraction (XRD) was used for analysis Chloriti Lapis and the XRD Fourier fingerprints were established. The dates were analyzed by fuzzy cluster and fingerprint similarity evaluation software to compare the similarity of samples. XRD fingerprint with 10 common peaks of 14 batches of Chloriti Lapis were established. The average, median coefficients of crystal lattice spacing d (A), peak position 2 theta, relative intensity value I/I0 (%) were all more than 0.95. And similarity( angle cosine value) were all more than 0. 97. There were small number samples differed from others. And obvious differences between the pre-and post-processing samples. This paper shows the powder XRD Fourier fingerprint can be used for appraisal and study of the Chloriti Lapis.

[Simultaneous determination of the five alkaloids in Rhizoma Coptidis by nonaqueous capillary electrophoresis].

A method for the simultaneous determination of berberine, palmatine, jatrorrhizine, magnoflorine and coptisine from Rhizoma Coptidis samples based on the nonaqueous capillary electrophoresis (NACE) mode has been developed. The effects of several important factors, such as nonaqueous solvents, running buffer system and its concentration and pH, separation voltage, temperature and detection wavelength, were investigated to acquire the optimum conditions. The optimum conditions for the separation were as follows: the selected running buffer was a methanol solution (pH 5.8) containing 40 mmol/L sodium acetate and 40 mmol/L ammonium acetate; the separation voltage was 25 kV; detection wavelength was set at 254 nm; the sample was injected at 5 kPa x 6 s and the column temperature was maintained at 20 degrees C. The analytes can be obtained good baseline resolutions in a 64.5 cm x 75 microm capillary (56 cm of effective length) within 20 min. The average recoveries of the established method were between 98.37% and 101.03%. The method is simple, accurate and reproducible, and can be used for the quality control analysis of Rhizoma Coptidis.

[Studies on NACE-DAD fingerprint of Cortex Phellodendri].

OBJECTIVE: To establish the analytical method for the fingerprint of Cortex Phellodendri by NACE-DAD and estimate the quality of Cortex Phellodendri from different habitats and species. METHODS: Based on the mode of nonaqueous capillary electrophoresis, 40 mmol/L sodium acetate and 40 mmol/L ammonium acetate methanol solution was selected for the buffer (pH 5.8). The separation voltage was 25 kV and detection wavelength was set at 210 nm. Berberine was used as a reference standard, the chromatographic fingerprint was determined. The data were analysed by Fuzzy Cluster and Fingerprint Similarity Evaluation Software to compare the similarity of samples. RESULTS: NACE-DAD fingerprints with 9 common peaks were established preliminarily. It was discovered that a small number of samples differed from others. Regarding to the fingerprints of Cortex Phellodendri and its processed products, there were obvious differences in the relative areas of common peaks. CONCLUSION: The method is reliable, accurate and can be used for quality control of Cortex Phellodendri.