[Determination of six compounds in Berberidis Cortex and comparative study of its different species].

In order to clarify the characteristic components of Berberidis Cortex,the preparative liquid chromatography and spectral analysis methods were used to separate and identify the unknown components in the water extract of Berberidis Cortex. Two compounds were isolated and identified as bufotenidine and ferulic acid 4-O-ß-D-glucopyranoside. They were both isolated for the first time from Berberidis Cortex and Berberis. In addition,an HPLC method was successfully established for simultaneously determination of six compounds in Berberidis Cortex,and chemometric methods were used to study the chemical differences among three main species of Berberidis Cortex. The results suggested that jatrorrhizine and bufotenidine are the main difference compounds among the three species.Compared with B. kansuensis and B. diaphana,B. vernae contains significantly more jatrorrhizine(P<0. 01),and the content of bufotenidine in B. vernae was significantly higher than that in B. kansuensis(P<0. 05). Considering these results,further research is necessary to reveal the pharmacological activities of bufotenidine and the pharmacodynamic differences between the three species. The results could provide a reference for quality control,the basic research on effective substances,and development of Berberidis Cortex.

[Determination of six alkaloids in six types of Coptidis Rhizoma pieces by RP-HPLC and spectrum-effect relationships with anti-diabetes pharmacodynamics data].

To establish a method for determining the contents of six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride) in six types of Coptidis Rhizoma pieces (crude pieces, ginger juice stir-fried pieces, vinegar stir-fried pieces, wine steamed pieces, wine stir-fried pieces, evodiae juice stir-fried pieces) by RP-HPLC, and explore the relationship with the curative effect of traditional Chinese medicine (TCM) and pharmacodynamics results. The chromatographic column was Welch XtimateTM C18 (4.6 mm×250 mm, 5 µm), with 0.1% triethylamine solution (adjust pH at 10 with ammonium bicarbonate and ammonia) as mobile phase A and acetonitrile as mobile phase B for gradient elution (0-15 min, 10%-25%B; 15-25 min, 25%-30%B; 25-40 min, 30%-45%B) at a rate of 1.0 mL•min⁻¹. The column temperature was set at 30 ℃, and the wavelength was set at 270 nm. The six alkaloids showed a good linear relationship within the range of 0.85-16.96 mg•L⁻¹ (r=0.999 7), 1.25-24.96 mg•L⁻¹ (r=0.999 9), 2.05-40.96 mg•L⁻¹ (r=0.999 9), 3.65-72.96 mg•L⁻¹ (r=0.999 9), 2.88-57.60 mg•L⁻¹ (r=0.999 8), and 13.25-264.96 mg•L⁻¹ (r=0.999 6) respectively. The average recoveries (n=9) of the six alkaloids were 102.4% (RSD 1.2%), 101.8% (RSD 1.3%), 100.3% (RSD 1.8%), 100.7%(RSD 1.8%), 101.2% (RSD 1.5%) and 97.90% (RSD 2.0%) respectively, and their average contents were 3.55, 4.49, 9.12, 19.17, 15.69, 62.56 mg•g⁻¹, respectively. This determination method was accurate and repeatable, which could be used for the content determination in six types of Coptidis Rhizoma pieces. Data analysis on contents determination and preliminary pharmacodynamics results was conducted by using principal component analysis (PCA) and hierarchical clustering analysis (HCA). The analysis results showed that three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces) had significant differences with crude pieces, and the wine steamed Coptidis Rhizoma pieces showed most difference with crude pieces especially, mainly related to triglyceride (TG) and fasting blood glucose levels (FBG) in serum. In addition, columbamine hydrochloride was most affected among the six alkaloids. Those three types of Coptidis Rhizoma pieces (wine steamed pieces, wine stir-fried pieces, and evodiae juice stir-fried pieces), had more advantages for "anti-diabetes" in TCM clinical application, especially in the treatment of diabetic hyperlipidemia.

[Study on quality standard of Berberidis Cortex].

In order to establish the quality standard of Berberidis Cortex and improve its quality control level, water, total ash, acid-insoluble ash and alcohol-soluble extract were determined according to procedures recorded in the Chinese Pharmacopoeia (2010 edition). The qualitative and quantitative analyses were performed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) methods. The results showed that TLC identification had a good resolution with clear spots. The water content was 8.39%-12.23%; total ash was 4.50%-9.96%; acid-insoluble ash was 0.10%-0.69%, and the alcohol-soluble extraction was 20.62%-37.13%. The average contents of magnoflorine, jatrorrhizine, palmatine, and berberine in Berberidis Cortex were 5.98%, 0.63%, 0.30%, 2.50%, respectively. It was concluded that the developed method was accurate and good in specificity, which can be used for quality control of Berberidis Cortex in the future.

[Studies on dosage-efficacy/toxicity relationship of anti-hypoxia effects and quality standard of Tibetan medicine Brassicea Radix extract].

To evaluate the efficacy and safety associated with anti-hypoxia effect and establish the quality standard for Brassicea Radix extract, the investigations of acute toxicity and subacute toxicity were carried out to preliminarily appraise the toxicity, and the models of normal pressure hypoxia, acute cerebral ischemia and sodium nitrite poisoning in mice were used to evaluate the effect of enhancing anoxia endurance. Then according to the methods described in the Appendix of Chinese Pharmacopoeia (2010 edition), the sulfuric acid-phenol method was applied to determine the content of polysaccharide, and the water, ash and insoluble matter in water inspections were carried out and the control medicinal herb was identified with the samples by qualitative TLC. The results indicated that ① the toxic effects (LD50) of mice was 56.73 g•kg⁻¹ by oral administration of Brassicea Radix extract, while Dm and Dn were respective 86.80 g•kg•d⁻¹ and 35.55 g•kg•d⁻¹;②the determined effective dosage of Brassicea Radix extract which could enhance anoxia endurance was 0.388 g•kg⁻¹â€¢d⁻¹; ③ the methods of TLC and the content of polysaccharide were established. The method of quality control has been recorded in Sichuan Province Standard for Tibetan Medicine, which is reliable, accurate and simple, with good reproducibility. Meanwhile, given the prominent effect on anti-hypoxia and good safety, it provided important basis for clinic safe and effective usage and the development of health products.

[Analysis and thinking on status of quality standard of ethnic medicine].

The establishing and improving of quality standards for ethnic medicine have close relation with the safety and efficacy of clinical medicine. It is significant to promote the modernization and industrialization of ethnic medicine. This paper summarized the present status of quality standards for ethnic medicine, and pointed out the deficiency of ethnic medicine; the origin of medicinal materials was not clear and varieties were confused; the safety research of mineral drug was weak and so on. Based on these, firstly, the research of quality standards for ethnic medicine should be under the guidance of ethnic traditional medical theory and clinical experience and strengthen the quality control and evaluation of safety and effectiveness for ethnic medicine. Further, much attention should be payed to protection of natural medicinal resources and sustainable utilization. Lastly, modern chemical analysis technology shouled be used during the research process on the basis of ethnic characteristics. It will promote the establishment and improvement of quality standards for ethnic medicine and will be benificial to modernization and industrialization development of ethnic medicine.

Quality evaluation and species differentiation of Rhizoma coptidis by using proton nuclear magnetic resonance spectroscopy.

Rhizoma coptidis, a broadly used traditional Chinese medicine, derives from the dried rhizomes of Coptis chinensis Franch, Coptis deltoidea C.Y. Cheng et Hsiao and Coptis teeta Wall. Quantitative determination of protoberberine alkaloids in R. coptidis is critical for controlling its quality. In this study, a rapid, simple and accurate quantitative (1)H NMR (qNMR) method was developed for simultaneous determination of berberine, jatrorrhizine, epiberberine, coptisine, palmatine and columbamine in R. coptidis from the three species. Method validation was performed in terms of selectivity, precision, repeatability, stability, accuracy, robustness and linearity. The average recoveries obtained were in the range of 96.9-102.4% for all the six alkaloids. In addition, the qNMR data were analyzed with analysis of variance (ANOVA), hierarchical clustering analysis (HCA) and principal component analysis (PCA), and the results showed that the contents of the active alkaloids have significant difference among the three species. Compared with the conventional HPLC approach, the proposed qNMR method was demonstrated to be a powerful tool for quantifying the six alkaloids due to its unique advantages of high robustness, rapid analysis time and no need of standard compounds for calibration curves preparation. These findings indicate that this method has potential as a reliable method for quality evaluation of herb medicines, especially for protoberberine alkaloid-containing ones.

Analgesic effect and mechanism of the three TCM-herbal drug-combination Tou Feng Yu pill on treatment of migraine.

It is well known that pain is one of the most important characteristics of migraine. Therefore, it is important and interesting to investigate the analgesic effect and mechanism of drugs which are used to treat migraine. Tou Feng Yu pill (TFY) is a traditional Chinese herbal medicine, consisting of the three Chinese herbal drugs Radix Angelicae dahuricae (Baizhi), Rhizome Ligustici (Chuanxiong) and Folium Camelliae sinensis (green tea) for the treatment of migraine. In this study, we found that TFY could significantly reduce the writhing times induced by acetic acid and licking foot response induced by formalin, and extend the writhing latent period. But the analgesic effect was not observed at hot-plate test. Meanwhile, experimental migrainous model induced by nitroglycerin was used to investigate the therapeutic effect of TFY. Compared with the control group, the levels of plasma calcitonin gene related to peptide (CGRP), serum nitric oxide (NO) and contents of brain dopamine (DA) in TFY administration groups were significantly decreased, and the levels of plasma endothelin (ET) and contents of brain 5-hydroxytryptamine (5-HT) and norepinephrine (NE) were remarkably increased, also the ratio of ET/NO was clearly corrected. Furthermore, the improving effect of behavior abnormality was observed in TFY administration rats. Meanwhile, isolated vascular ring test indicated that TFY had an significant effect on vasomotion, and antagonize vasospasm; moreover TFY also could increase cerebral blood flow (CBF) and reduce cerebrovascular resistance index (RI) in normal rabbits, which verified the effect of TFY on vasomotion and abnormal hemorheology. All the results indicate that TFY has an effective therapeutical action on migraine, which may be related to three aspects as following: firstly, adjusting the level of neurotransmitters, neuropeptides and vasoactive substances, relieving neurogenic inflammation; secondly, improving vasomotion, increase cerebral blood flow, then improving hemorheology; finally, increasing pain threshold, relieving or preventing headache. These findings provide additional pharmacological information and may contribute for the further study and use of TFY as a phytomedicine.

[Studies on quality standard of Yifu ointment].

OBJECTIVE: To develop a method for the determination of Chonglou saponin I and Chonglou saponin II in Yifu ointment. METHOD: The chromatographic separation was performed on Hypersil ODS2 C18 column (4.6 mm x 150 mm, 5 microm) and IBBM-612111 ODS C18 guard column. Acetonitrile-0.02% phosphoric (43:57) as mobile phase. The flow rate was 1 mL min(-1) and column temperature was set at 40 degrees C. The UV detection wavelength was set 203 nm. RESULT: Chonglou saponin I showed a good linear relationship at a range of 0.1024-3.2 microg, r =0.9998, the average recovery was 97.4%, and RSD was 1.8% (n = 6); Chonglou saponin II showed a good linear relationship at a range of 0.064-2.0 microg, r = 0.9999, the average recovery was 101.4%, and RSD was 1.0% (n =6). CONCLUSION: The method is accurate with the good reproducibility and can be used for the quality control of Yifu Ointment.

[Quality standard for Xindi soft capsule].

OBJECTIVE: To establish the quality control standard of Xindi soft capsule. METHOD: Quercetin, kaempferol and isorhamnetin were isolated by TLC with chloroform-ethyl formate-formic acid (5:4:1). The chromatographic separation was performed on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm). Acetonitrile-water-phosphoric (30:70:0.1) as mobile phase. The flow rate was 1 mL x min(-1) and column temperature was set at 40 degrees C. The UV detection wavelength was set at 254 nm. RESULT: Quercetin, Kaempferol and Isorhamnetin could be identified by TLC. Quercetin showed a good linear relationship at a range of 0.412-1.648 microg, r = 0.999 9, the average recovery was 96.8%, and RSD was 0.9% (n = 6). Kaempferol showed a good linear relationship at a range of 0.021-0.083 microg, r = 0.999 8, the average recovery was 96.9%, and RSD was 2.0% (n = 6). Isorhamnetin showed a good linear relationship at a range of 0.183-0.732 microg, r = 0.999 9, the average recovery was 97.1%, and RSD was 1.6% (n = 6). CONCLUSION: The method is accurate with the good reproducibility and can be used for the quality control of Xindi soft capsule.