[Qualitative and quantitative analysis on triterpenoids in Ligustri Lucidi Fructus].

Triterpenoids are one of the most active constituents in Ligustri Lucidi Fructus, but only oleanolic acid has been mostly studied. In recent years, a growing number of studies have shown that other triterpenes from Ligustri Lucidi Fructus also have various biological activities, so it is necessary to build up a detailed profile of the triterpenoids in Ligustri Lucidi Fructus. The chromatographic separation was performed on a C_(18) column(4.6 mm×250 mm, 5 µm) with mobile phase of methanol-acetonitrile-0.2% formic acid for gradient elution. The detection wavelength was set at 210 nm, with a flow rate of 0.5 mL·min~(-1), and the column temperature of 25 ℃. The HPLC fingerprint of triterpenoids in Ligustri Lucidi Fructus was built by testing 21 batches of samples from different sources. The structures of the total 15 common chromatographic peaks were elucidated with UHPLC-ESI-Orbitrap-MS/MS technique and six of them were identified as tormentic acid, pomolic acid, maslinic acid, botulin, oleanolic acid and ursolic acid by comparison to the reference substances. Under the same chromatographic condition, four main triterpenes(podocarboxylic acid, hawthorn acid, oleanolic acid and ursolic acid) were quantified and the results of system adaptability and methodology investigation all met the requirements of content determination. Meanwhile, with oleanolic acid(A) as the internal reference substance, quantitative analysis of multi-components by single marker(QAMS) method was used to analyze the above four components. The relative correction factor of oleanolic acid(B), hawthorn acid(C) and ursolic acid(D) to oleanolic acid was f_(B/A)=1.12, f_(C/A)=1.02 and f_(D/A)=0.88, respectively, and the relative retention values of these three to oleanolic acid was RRV_(B/A)=0.46, RRV_(C/A)=0.70 and RRV_(D/A)=1.03, respectively. The contents determined by two methods were similar. In conclusion, the method built in this paper is proved to be simple, reliable and specific for the simultaneous qualitative and quantitative analysis of the triterpenoids in Ligustri Lucidi Fructus, which can lay foundation for further assays of the triterpenoids in Ligustri Lucidi Fructus and the relative products.

[Qualitative and quantitative analysis on non-triterpenoids in Ligustri Lucidi Fructus].

In order to improve the quality control level of Ligustri Lucidi Fructus(LLF) and to explore the changes of chemical components after processing,the HPLC method for fingerprint and simultaneous determination of the major polar components in LLF were established. The octadecylsilane bonded silica gel was used as the stationary phase,with acetonitrile as the mobile phase A and0. 2% formic acid as the mobile phase B in a gradient elution procedure at a flow rate of 1. 0 m L·min-1. The detection wavelength was set at 280 nm and the column temperature was 25 ℃. There were 22 common peaks,20 of which were selected from the fingerprint of LLF and its wine-steamed product,respectively,and 14 chromatographic peaks were identified with reference substances. With the same chromatographic conditions,seven components were quantitatively analyzed and the results of system adaptability and methodology investigation all met the requirements of content determination. Compared with the crude LLF,the content of 5-hydroxymethyl furfural and salidroside significantly increased in wine-steamed LLF,while the contents of iridoid glycosides generally decreased. The method provided a basis for quality control of LLF and its processed products as well as the related preparations.

Effects of benzo(a)pyrene on the contractile function of the thoracic aorta of Sprague-Dawley rats.

OBJECTIVE: To evaluate the possible vascular effects of an environment carcinogen benzo(a)pyrene (BaP). METHODS: The cytotoxicit of BaP and rat liver S9 (0.25 mg/mL)-activated BaP were examined by MTT assay. Thoracic aortic rings were dissected from Sprague-Dawley rats. Contraction of aortic rings was induced by 60 mmol/L KCl or 10(-6) mol/L phenylephrine (PE) in an ex-vivo perfusion system after BaP (100 µmol/L) incubation for 6 h. [Ca(2+)](i) was measured using Fluo-4/AM. For in-vivo treatment, rats were injected with BaP for 4 weeks (10 mg/kg, weekly, i.p.). RESULTS: BaP (1-500 µm) did not significantly affect cell viability; S9-activated BaP stimulated cell proliferation. BaP did not affect the contractile function of endothelium-intact or -denuded aortic rings. BaP did not affect ATP-induced ([Ca(2+)](i)) increases in human umbilical vein endothelial cells. In BaP-treated rats, heart rate and the number of circulating inflammatory cells were not affected. Body weight decreased while blood pressure increased significantly. The maximum aortic contractile responses to PE and KCl and the maximum aortic relaxation response to acetylcholine were significantly decreased by 25.0%, 34.2%, and 10.4%, respectively. CONCLUSION: These results suggest, in accordance with its DNA-damaging properties, that metabolic activation is a prerequisite for BaP-induced cardiovascular toxicity.

[Preliminary study on variation pattern of Cistanche deserticola].

OBJECTIVE: Through analyze of shape, structure and content of effective components of Cistanche deserticola, the variation of "you cong rong" is discussed. METHOD: The color surface, texture, the size of pith and the arrange of vascular bundle was observed with freshed, dried drugs and the transverse section, the effective component of echinacoside and acteoside was analyzed with hplc and the amylose was analyzed with ultraviolet spectrophotometry. RESULT: The variation of "you cong rong" is obvious different with the common drug in shape and structure, the content of echinacoside is 2.5 times, acteoside is 3.8 times, amylose yield is 1.6 times and the content is 2 times of the common drugs. CONCLUSION: "you cong rong" is an independent variation pattern of C. deserticola, it has no relationship with the ecological and geographical conditions.

[Primary study on shapes of fruits and germination characters of seeds of Radix et Rhizoma Rhei].

OBJECTIVE: To explore the difference of the shapes of fruits and germination characters of seeds of Rheum palmatum, R. tanguticum and R. officinale. METHOD: The seeds of three Rheum species including wild and cultivation one were collected from different regions. Character of the fruits appearance were observed. The purification, weights per thousand seeds, content of moisture, seed vigor and different germination rates of Radix et Rhizoma Rhei were measured. The germination rates of the seeds of R. palmatum were determined under different temperature and hormone treatment . RESULT: It was markedly variant in shapes of the fruits, rates of germination and all quality characters of the three category of Radix et Rhizoma Rhei. As reported, the size of the fruit and seed and weights per thousand seeds of R. officinale was the biggest. The seed vigor and germination rate of R. palmatum caltivated in Ming county of Gangsu were 95.7% and 94% respectively. The rate of seed vigor was closely correlated with the rate of germination for other species. Results indicated that temperature had some effects on the germination, and there were obvious difference in the active effect of gibberellin, kinetin and 6-benzyl aminopurine on the germination of the seed. Our experimental results showed that the kinetin concentration of 50 x 10(-6) g x L(-1) had the most active effect, and its rates of germination increase 18 percent than the control one. CONCLUSION: Significant differences exist in the shapes of fruits, rates of germination of the seeds of Rheum, which are affected by the factors of temperatures and hormones. The result of this study provides scientific evidences for identify, growth and cultivation of Rheum.

[The determination of echinacoside and acteoside in herbs of Cistanche tubulosa].

OBJECTIVE: To analyse the echinacoside and acteoside content of Cistanche tubulosa, collected from different hosts and different size of the cultivated, which is compared to the wilding by RP-HPLC method. METHOD: An Agilent Eclipse XDB-C18(4.6 mm x 250 mm, 5 microm) column was used and a mixture of methanol-acetonitrile-1% acetic acid (15:10:75) was used as the mobile phase at a flow rate of 0.6 mL x min(-1). The column temperature was 30 degrees C and the UV detection wavelength was 334 nm. RESULT: The calibration curves of echinacoside and acteoside were in good linearity over the range of 0.904-9.04 microg (r = 0.999 9), and 1.27-12.7 microg (r = 0.999 9) respectively and the average recoveries of echinacoside (and acteoside) were 98.9% (n = 5, RSD 1.9%), and 97.0% (n = 5, RSD 0.97%). CONCLUSION: The method is simple, quick, acurate. In all of the samples, the contents of echinacoside is markedly more than that of acteoside, the content of the two active component in the wilding is higher than that in the planting. The content of sample in the different sizes gradually increase from the big to the small, and the contents of samples collected from the different hosts vary markedly. These results are useful for the quality evaluation of medicinal materials of C. tubulosa.

[Study on genetic diversity of herba Cistanches by RAPD].

OBJECTIVE: To determine the genetic diversity of Cistanche species. METHOD: Two populations of Cistanche deserticola and four populations of C. tubulosa were analyzed by random amplified polymorphic DNA (RAPD) markers. RESULT: A total of 76 and 87 loci were amplified using 10 random primers each other. The average percentage of polymorphic loci of C. deserticola was 47.37%. The PPL were 39.47% and 35.53% for two populations. Average Nei's gene diversity was 0.1358, Shannon' s genetic diversity was 0.2072, and Gst was 0.2546. The average PPL of C. tubulosa was 27.59%. It was 19.54% to 25.29% in different populations and Andi'er population had the highest. Average Nei's gene diversity was 0.0823, and Shannon' s genetic diversity was 0.125 8, Cst was 0.175 5. CONCLUSION: The diversity of Cistanche deserticola is higher than that of C. tubulosa, but both has differentiation among populations, C. deserticola has already separated itself into two different ecotypes.