[Application of FTIR and Active Ingredients Quantitative Analysis on Quality Control of Dai Medicine Alstonia scholaris (L.) R. Br.].

FTIR fingerprint of the leaves and immature stems of Alstonia scholaris (L.) R. Br. was established as a content determination method for the detection of picrinine, ursolic acid and oleanolic acid. Different medicinal parts were identified based on principal component analysis, while exploring the influence of immature stems for the leaves and the application of FTIR and HPLC in the Dai quality control in order to speed up the pace of Dai medicine modernization. Infrared spectroscopy of different batches samples were collected and the data was preprocessed as to automatic baseline correction, smooth, ordinate normalization, second order derivative, and then to PCA, all the datum in triplicate. For content determination of picrinine, mobile phase was acetonitrile (40) water (contain 0.1% ammonia water) (60) and the wavelength was set at 287 nm. For ursolic acid and oleanolic acid, the mobile phase was mixture (12∶88) of 0.1% formic acid in water (A) and methanol (B). Wavelength was 210 nm. As the results, the original spectrum difference was not obvious for leaves and stems. Pretreatment spectroscopy had a significant variation on absorption peak number and intensity in 3 000～2 800 and 1 800～500 cm(-1). The results of PCA showed that, the leaves and stems were separated; in addition the difference of different batches leaves was bigger than the stems. The mean contents of picrinine, ursolic acid and oleanolic acid in leaves were 0.79，8.47，7.51 and 0.21，1.78，1.67 mg·g(-1) in stems, respectively. The content of ursolic acid and oleanolic acid is higher than picrinine, but ursolic acid and oleanolic acid content had no obvious difference. Mean content of three ingredients in leaves is much higher than in stems. Picrinine content in leaves was 3.8 times of immature stems, ursolic acid and oleanolic acid content were 5.1 and 4.2 times of immature stems, respectively. The variety of picrinine content in different batches samples was biggest, ursolic acid and oleanolic acid content was relatively stable. The overall quality of leaves has an obvious difference compared with the immature stems, so the leaves of A. scholaris mix with immature stems could not be as Dai medicine in Dai clinic. Infrared spectroscopy combined with chromatography can quickly identify different medicinal parts and evaluate overall quality of Dai medicine, which can apply to quality control of Dai medicine.

[Distinguish and Quality Estimation of the Leaves of Alstonia scholaris (L.) R. Br. from Different Harvest Time Based on the UV-Vis·FP and HPLC·FP].

UV-Vis and HPLC fingerprint of different harvest time of the leaves of Alstonia scholaris (L.) R. Br. were establish the for identification and quality evaluation to promote the development of Dai Medicine modernization. The optimal extraction condition was used to obtain UV - vis data of different harvest time which were deducted background and eight spot smooth, were collected to make the principal component analysis in SIMCA-P(+)11.5, identifying the samples quickly with the first three principal component three-dimensional diagram. The HPLC fingerprint were obtained with Agilent ZORBAX Eclipse XDB C18 (250×4.6 mm, 5 µm) chromatographic column with the mobile phase of acetonitrile (B) - water (contain 0.1% formic acid) (A) for gradient elution (0～5 min, 5% B; 5～35 min, 5% B→26% B; 35～40 min, 26% B→56% B). The wavelength was set at 287 nm and the column temperature was maintained at 30 ℃. The flow rate was 1.0 mL·min-1 and the injection volume was 7 µL. The HPLC fingerprint of different harvest time of the leaves of Alstonia scholaris (L.) R. Br. was analysised by cluster analysis to quality evaluation. Research findings showing: (1) The UV-Vis spectrogram of different harvest time of the leaves of Alstonia scholaris (L.) R. Br. were divided into three parts according to the absorption peak position and amplitude of variation. The first was 235 to 400 nm, the second was 400 to 500 nm, and the third was 500 to 800 nm. In the first part, absorption peak were focused on 270, 287 and 325 nm, which can reflect the fingerprint character for the high absorbance and amplitude of variation. Absorption peak were distributed in 410 and 464 nm in the second part, absorbance and amplitude of variation were lower than the first part. There was a bigger absorption peak at 665 nm in the third part, but the absorbance had no difference. The UV-Vis data of different harvest time were gathered to make the principal component analysis, the result was that the samples of same month were concentrated distribution, but different month samples were dispersed distribution. (2) HPLC fingerprint were divided into three categories through hierarchical cluster analysis, 3, 4, 5 and 7 month were the first category, 6, 8, 9 month samples were second category, the others were third category. Chemical composition and content of the same category samples were similar, but the different category samples had a obvious difference, more important is that the third category samples content was the highest. Combining UV-Vis FP and HPLC FP can identify and evaluate quickly the samples of different harvest time of the leaves of Alstonia scholaris (L.) R. Br. The optimal harvest time of Alstonia scholaris (L.) R. Br. was from October to next February, which was the coldest season in the Dai calendar.

[Comparison of rattan microscopic characteristics of four Dai medicinal materials from Uncaria genus].

OBJECTIVE: To establish microscopic identification standards for four species from Uncaria genus. METHODS: The microscopic characteristics of transvers section and powder of these four species rattan were observed. RESULTS: As for these four species from Uncaria genus, the microscopic identification of rattan transvers section could be made by the existence of cork cambium and pericycle in the cortex, and the location of phloem fiber and stone cells, while the powder identification could be made according to the existence of scalariform vessel and calcium oxalate crystal, and the quantity of stone cells. CONCLUSION: The microscopic characteristics of rattan can provide information for identification of these four species from Uncaria genus.

Sulfur-containing amides from Entada phaseoloides.

To study the chemical constituents of the Entada phaseoloides (L.) Merr., seeds of Entada phaseoloides were extracted with 70% ethanol at room temperature. Isolation and purification were performed by silica gel, reversed-phase silica gel column chromatography and semi-preparative HPLC. Structures of the pure compounds were established on the basis of spectral analysis. Four sulfur-containing amide compounds were isolated from the n-BuOH-soluble fraction and identified as entadamide A-beta-D-glucopyranosyl-(1-->3)-beta-D-glucopyranoside (1), entadamide A (2), entadamide A-beta-D-glucopyranoside (3) and clinacoside C (4). Compound 1 is a new compound. Compound 4 is isolated from the genus Entada for the first time.

Two new triterpenoids from Lycopodium obscurum L.

Two new onoceranoid triterpenoids, (3 alpha,8 beta,14 alpha,21 beta)-26,27-dinoronocerane-3,8,14,21-tetrol (1) and 26-nor-8 beta-hydroxy-alpha-onocerin (2), were isolated from Lycopodium obscurum L. Their structures were elucidated on the basis of spectroscopic analyses.

[Study on acute toxicity and animal gastrointestinal activity of crude and processed products of Entada phaseoloides].

OBJECTIVE: To study the acute toxicity of the crude and processed products of Entada phaseoloides and their effects on gastrointestinal movement in mice. METHODS: Using the method of intragastric administration, to observe the acute toxicity of the crude and processed products of Entada phaseoloides in mice and determine their LD50. With the methods of charcoal propulsion of small intestine and methyl orange colorimetry of gastric emptying, to study the impact of the crude and processed products of Entada on gastrointestinal movement in mice. RESULTS: The oral LD50 of crude Entada phaseoloides, No. 1 and No. 2 processed products of Entada phaseoloides in mice were 27.17, 35.13, 42.18 g/kg body weight. Crude and processed products of Entada phaseoloides can significantly promote the enteric propulsion of normal mice, and can significantly counteract the depressing status induced by atropine, but have no influence on the overactive status induced by neostigmine. The high, middle and low-dose of groups showed significant inhibition of the gastric emptying in normal mice. CONCLUSION: Processed Entada phaseoloides showed effects on the enteric propulsion of normal and depressing mice, can restrain the gastric emptying under normal mice, but its safety is better than crude Entada phaseoloides.

[Study on microscopic observation and TLC identification of Dai medicine "Guomaguo" (Spondias pinnata)].

OBJECTIVE: To study identification methods of Dai medicine" Guomaguo", the fruit of Spondias pinnata. METHODS: Characteristic, microscopic observation and TLC idertification were used to authenticate this crude drug. RESULTS: The characters of the cross section, powder and TLC of the drug were reported, and the relevant drawings of the tissue, powder and TLC of this ethnomedicine were drawn. CONCLUSION: These results can supply evidences for the identification of the ethnomedicine in its exploitation and utilization.

[Study on pharmacognostic identifcation of Cassia siamea].

OBJECTIVE: To establish methods for identification of Dai medicine, the heartwoods and leaves of Cassia siamea. METHODS: Macroscopic, microscopic observation and TLC technique were used to anthenticate this crude drug, and the identification characteristics were studied. RESULTS: Macroscopic and microscopic identification methods and TLC characters of the ethnomedicine were re-ported, and the simple and detailed drawings of the transections of the heartwoods and leaves, as well as microscopic drawings of their powders were drawn in this paper. CONCLUSION: The results can serve as evidence for identification of the ethnomedicine in the utilization.