| Elsholtzia rugulosae Folium is the dry leaf with twig of a lamiaceae plant called Elsholtzia rugulosa Hemsl. Elsholtzia rugulosae Folium is cool in nature and has an acrid flavor. As a commonly used ethnic folk drug, it is effective in dispelling wind, relieving exterior syndromes, promoting diuresis and digestion, and relieving stasis, and has been used for a long time by various ethnic groups. In this paper, the pharmacognosy of Elsholtzia rugulosae Folium is studied systematically, to support the "study of the improvement of quality standards of local conventional Chinese medicine in Sichuan Province". Based on the pharmacognosy study and the technical requirements for setting quality standards for Chinese medicine, the quality standards of Elsholtzia rugulosae Folium have been proposed, part of which have been included in the Sichuan Standards for Chinese Medicine (2010Edition). In addition, the GC-MS analysis,GC fingerprint chromatogram and pharmacology of Elsholtzia rugulosae Folium have been studied, providing scientific basis for further quality control and clinical practice. The main study contents and results are as follows:Morphological and histological studies of Elsholtzia rugulosae Folium have been carried out in detail, through which the main identified characteristics in terms of the original plant, properties and tissue structure have been found out. The morphology of the original plant is identified in such aspects as the insertion modes and shapes of leaves and flowers, hairs on leaves and shapes of fruits; the properties are identified mainly by hairs over twigs, shape and color of leaves and hairs over the upper and lower surfaces of leaves; regarding to the tissue structure, the cross section of the stem is identified in terms of glandular hairs, non-glandular hairs, collenchyma, vascular bundle type, etc.; leaves are identified in terms of glandular hairs, non-glandular hairs, palisade tissue, spongy tissue, collenchyma, vascular bundle type, cell shape in the upper epidermis, etc.; powder is identified mainly from cell debris in the upper epidermis of leaves, non-glandular hairs, glandular hairs, glandular scales, vessels, fibers, etc. The thin-layer chromatography (TLC) method to identify the composition of volatile oil in Elsholtzia rugulosae Folium has been established. With cyclohexane-ethyl acetate (10:1) as the developing solvent, the separation effect is good, the Rf value after developing is moderate, and the spots are clear. The TLC tests of samples of Elsholtzia rugulosae Folium from different production areas shows that in the color spectrum of the samples, spots of the same color are displayed at places corresponding to those in the color spectrum of the control group. The intensity of spot color indicates the difference in content.The moisture, total ash and acid-insoluble ash in11batches of Elsholtzia rugulosae Folium from different production areas have been inspected with the method stated in the appendix of the first volume of Chinese Pharmacopoeia (2010edition). The standard limits have been proposed according to experimental data, commodity conditions and actual production:the moisture, total ash and acid-insoluble ash shall not exceed12.0%,9.0%and1.5%respectively.The method to determine the extract from Elsholtzia rugulosae Folium has been established with aqueous as the solvent and the hot dipping process applied. According to the experimental data in11batches of Elsholtzia rugulosae Folium from different production areas and the actual production, it has been proposed that the limit of the content of extract from Elsholtzia rugulosae Folium shall not be lower than18.0%.The content of volatile oil in Elsholtzia rugulosae Folium from different production areas has been determined with the method stated in the appendix of the first volume of Chinese Pharmacopoeia (2010edition). The content of volatile oil in11batches of samples is0.53%-1.88%and it is temporarily determined that the content of volatile oil shall not be less than0.60%.The method to determine the content of total flavone in Elsholtzia rugulosae Folium has been established with the UV-Vis spectrophotometry. The results from11batches of Elsholtzia rugulosae Folium show that the contents of total flavone in Elsholtzia rugulosae Folium from different production areas are significantly different, ranging from6.29%to17.39%. This may be associated with the soil nature, local climate and other external conditions in the different growing environments of the crude drugs.GC-MS analysis has been performed for volatile oil in three batches of Elsholtzia rugulosae Folium, the principle components are ketone,alcohol, aldehyde and carbene, followed by acid,ester and alkane. The No.1sample has identified76kinds of components,with the principal components being elsholtzia ketone, piperitone, benzaldehyde and linalool; the No.2sample has identified31kinds of components, with the principal components being elsholtzia ketone, beta-caryophyllene and dehydro-elsholtzia ketone; and the No.3sample has identified44kinds of components, with the principal components being piperitone,linalool,benzaldehyde and elsholtzia ketone.GC fingerprint chromatogram of volatile oil in Elsholtzia rugulosae Folium from different production areas have been studied for the first time, and the common mode of fingerprint chromatogram of Elsholtzia rugulosae Folium has been established preliminarily, where there are12characteristic peaks.Anti-inflammatory and in-vitro antibacterial experiments have been performed for Elsholtzia rugulosae Folium for the first time. The anti-inflammatory experiment shows that the aqueous extracts of Elsholtzia rugulosae Folium, being15g/kg,7.5g/kg and3.75g/kg, have significant inhibitory effect on ear swelling of mice induced by dimethylbenzene,indicating that Elsholtzia rugulosae Folium has good anti-inflammatory effect; the results of in-vitro antibacterial experiment show that the aqueous extracts and alcohol extracts of Elsholtzia rugulosae Folium have strong inhibitory effect on pseudomonas aeruginosa, escherichia coli, duck salmonella, staphylococcus aureus and staphylococcus epidermidis with the bacterial quantity of104-105CFU/ml,and the aqueous extracts is more effective than alcohol extracts. The acute toxicity test shows that the maximum tolerance dose of the aqueous extracts of Elsholtzia rugulosae Folium is320g crude drug per kilogram, corresponding to427times the clinical dose of an adult per day, indicating that it is safe for clinical use, with little adverse effect.Scientific basis has been provided for the determination of quality standards of Elsholtzia rugulosae Folium in the Sichuan Standards for Chinese Medicine (2010Edition) through the systematic pharmacognosy study of Elsholtzia rugulosae Folium, with the purpose of ensuring stable and controllable quality, and safe and effective. It sets the foundation for further indepth study, and has practical meaning for the development and utilization of ethnic folk drug resources of our country. |
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