ABSTRACT
A quantitative analysis method for ten principal components (phenylethanol, iridoids and triterpenes) of raw Ligustri Lucidi Fructus and its wine-steamed product was developed using liquid chromatography tandem triple quadrupole mass spectrometry (LC-QQQ-MS) to study their pharmacokinetic behavior in vivo. The results of methodological investigation were in accord with the criteria of biological analysis. After a single administration to rats of the water extracts of Ligustri Lucidi Fructus and its wine-steamed product, the plasma concentration of each component at different time points was measured and the pharmacokinetic parameters were determined. The AUC0-24 h and Cmax of the phenylethanol components (salidroside, tyrosol, hydroxytyrosol) were the greatest, suggesting that these components are the main pharmacological substances of Ligustri Lucidi Fructus. In addition, the tmax values of the eight major components were even lower with administration of the wine-steamed product, suggesting that these components are rapidly absorbed. However, the tmax values of specnuezhenide and oleanolic acid were greater with administration of the wine-steamed product, indicating that these two components were more slowly absorbed. A secondary peak phenomenon of tyrosol and hydroxytyrosol were observed in two sample groups, whereas the secondary peak phenomenon of salidroside occurred only with the wine-steamed product. This result suggests that the effect of wine-steamed product could persist for a long period. Meanwhile, the relative bioavailability of specnuezhenide and oleanolic acid was greater than 100% with administration of the wine-steamed product, consistent with the Traditional Chinese Medicine theory of the wine-steamed product being more effective than the raw material. The results reveal the different pharmacokinetic parameters and relative bioavailability of each component of Ligustri Lucidi Fructus and its wine-steamed product, and also demonstrate the variation and correlation of various components in vivo and in vitro, providing an experimental basis for the selection of quality control indexes, mechanisms of processing and the metabolic rule in vivo of Ligustri Lucidi Fructus. These experiments were approved by the Ethics Committee of Institute of Basic Theory for Chinese Medicine, China Academy of Chinese Medicine Science.
ABSTRACT
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.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fruit , Ligustrum , Tandem Mass Spectrometry , TriterpenesABSTRACT
Qingjin Huatanpang， first contained in Yixue Tongzhi， was composed of eleven medicinal materials of Scutellariae Radix， Gardeniae Fructus， Fritillariae Thunbergii Bulbus， Mori Cortex， Trichosanthis Semen Tostum， Citri Exocarpium Rubrum， Platycodonis Radix， Ophiopogonis Radix （core removed）， Anemarrhenae Rhizoma， Poria and Glycyrrhizae Radix et Rhizoma. It is a classic prescription created by YE Wen-ling in Ming dynasty for treating pulmonary disease with phlegm-heat obstructing lung syndrome. With the significant functions of clearing heat and moistening lung， reducing phlegm and relieving cough， it has been included in the "Classic Catalogue of Ancient Classics （First Batch）". Modern pharmacological studies have shown that Qingjin Huatantang has multiple activities such as relieving cough and eliminating phlegm， anti-inflammatory， bronchodilation， and immunoregulatory， and now it is commonly used for treating infectious lung diseases， such as acute exacerbation of chronic obstructive pulmonary disease， community acquire pneumonia， bronchiectasis， acute and chronic bronchitis in a form of its modified prescription or its combined use with western medicine， consistent with the clinical application in ancient times. According to the literatures on the study of Qingjin Huatantang published in recent years， this paper summed up the historical evolution， compatibility analysis， chemistry constituents， quality control， advances in pharmacology research， and clinical uses， which can provide theoretical and experimental data reference for further research and development， and proposed to establish a biological activity assay for quality control based on the pharmacological effect such as immunoregulatory activity， which can improve its quality control method and provide a reference for other famous classical formulas.
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The commodity specification and grade is a symbol of quality of Chinese medicinal materials,a standard to measure and evaluate the quality of Chinese medicinal materials,and an important part of the quality control system of Chinese medicinal materials.This article reviewed relevant research literature of commodity specification and grade of Chinese medicinal materials,including the formation and policy support of specification and grade.It analyzed the specific research examples of the commodity specification and grade of Chinese medicinal materials according to the classification of medicinal parts,and summarized the research conditions of the commodity classification at present stage.Aiming at the current research difficulties and the lack of unified standards in specification and grade of Chinese medicinal materials,some advice including research objects,methods,management,publicity,promotion and application are put forward,so as to provide references for the research on the commodity specification and grade of Chinese medicinal materials.
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Classical traditional Chinese medicine(TCM) excellent prescriptions is an outstanding representative in prescriptions of traditional Chinese medicine,clinical medicine effective tool,and the essence of traditional culture of Chinese medicine.The research and development of classical TCM excellent prescriptions has attracted great attention from group in the whole industry chain of TCM,a series of guiding suggestions and targeted literature laid a good foundation for research and development of classical TCM excellent prescriptions.In this paper,based on the CNKI database,literature about suggestions or enlightenments of classical TCM excellent prescriptions was sorted and refined in order to build the research outline of classical TCM excellent prescriptions,and it was concluded that literature research was the premise,medicine/decoction pieces with good quality was the foundation,standard substance was the core,high quality was the guarantee and preferential policies were the impetus,and related discussions were carried out.All these were expected to promote the research of classical TCM excellent prescriptions and development of related industries.
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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.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Fruit , Chemistry , Furaldehyde , Glucosides , Iridoid Glycosides , Ligustrum , Chemistry , Phenols , PhytochemicalsABSTRACT
This study was designed to establish a multiplex allele-specific polymerase chain reaction method for simultaneous identification of Dendrobium huoshanense, D. officinale and D. devonianum, which may resolve identification problems of caulis dendrobii. Internal transcribed spacer sequences and trnL-trnF sequences of the Dendrobium species were aligned by BioEdit software, then specific SNPs of the three species were analyzed for designing allele-specific primers and the multiplex allele-specific PCR reaction system was established. The different origin of Dendrobium huoshanense, D. officinale and D. devonianum was amplified and identified by the sizes of respective band. The results showed that 584 bp, 397 bp and 211 bp bands could be amplified by D. devonianum, Dendrobium officinale and Dendrobium huoshanense respectively, when the annealing temperature was 61 ℃ and the number of cycles was 35. The limit of detection (LOD) of D. devonianum and D. huoshanense were both 1.2 ng, while D. officinale was low than 0.24 ng. The detection limit of adulterates in D. devonianum, D. devonianum and D. huoshanense mixture sample was 1%, 1% and 5% respectively. This result suggests that the method of multiplex allele-specific PCR is useful to identify D. huoshanense, D. officinale and D. devonianum is accurate and specific.
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<p><b>OBJECTIVE</b>To evaluate the possible vascular effects of an environment carcinogen benzo(a)pyrene (BaP).</p><p><b>METHODS</b>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.).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>These results suggest, in accordance with its DNA-damaging properties, that metabolic activation is a prerequisite for BaP-induced cardiovascular toxicity.</p>
Subject(s)
Animals , Humans , Male , Rats , Aorta , Benzo(a)pyrene , Pharmacology , Calcium , Metabolism , Endothelial Cells , Metabolism , VasoconstrictionABSTRACT
<p><b>OBJECTIVE</b>Through analyze of shape, structure and content of effective components of Cistanche deserticola, the variation of "you cong rong" is discussed.</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>"you cong rong" is an independent variation pattern of C. deserticola, it has no relationship with the ecological and geographical conditions.</p>
Subject(s)
Amylose , Chemistry , Chromatography, High Pressure Liquid , Cistanche , Chemistry , Drugs, Chinese Herbal , Chemistry , Glucosides , Chemistry , Glycosides , Chemistry , Phenols , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To explore the difference of the shapes of fruits and germination characters of seeds of Rheum palmatum, R. tanguticum and R. officinale.</p><p><b>METHOD</b>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 .</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>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.</p>
Subject(s)
Benzyl Compounds , Fruit , Germination , Gibberellins , Pharmacology , Kinetin , Pharmacology , Plant Growth Regulators , Pharmacology , Plants, Medicinal , Physiology , Purines , Rheum , Physiology , Seeds , Physiology , Species Specificity , TemperatureABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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%).</p><p><b>CONCLUSION</b>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.</p>
Subject(s)
Breeding , Chromatography, High Pressure Liquid , Methods , Cistanche , Chemistry , Physiology , Glucosides , Glycosides , Host-Parasite Interactions , Phenols , Plants, Medicinal , Chemistry , Physiology , Tamaricaceae , Classification , ParasitologyABSTRACT
<p><b>OBJECTIVE</b>To determine the genetic diversity of Cistanche species.</p><p><b>METHOD</b>Two populations of Cistanche deserticola and four populations of C. tubulosa were analyzed by random amplified polymorphic DNA (RAPD) markers.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>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.</p>