Study On Extraction And Determination Methods Of The Active Ingredients From Chinese Herbal Drugs Compound Transdermal Patch

The chinese herbal drugs compound transdermal patch, which is a pure Chineseherbal drugs compound prescription for treating mastitis of dairy cows (Patent No.: ZL201010132973.2), contains Chinese Goldthread Rhizome, Indian Kalimeris Herb and Root,Hibiscus mutabilis L, Baikal Skullcap Root, Philippine Violet Herb and other ingredients.Its effects are anti-bacterial, anti-inflammatory, detoxification and promoting breast bloodcirculation. The main components are berberine chloride, baicalin and rutin in theprescription. Data reported that the Indian Kalimeris Herb.and Root and Philippine VioletHerb also contain flavonoids. The method for simultaneous determination of three kinds ofthe main active ingredients (berberine chloride, baicalin and rutin) was established in orderto determine the contents of the effective ingredients in the compound prescription, whichprovided the basis for developing the quality standards of the Chinese herbal drugscompound preparation.The laboratory extraction method of the Chinese drugs preparation was the Soxhletextraction method which was time-consuming and multiple steps. The orthogonal test wasused to find out the optimum ultrasonic extraction process for the transdermal patch inorder to adapt to the need to further expand the production. The gross peak areas and thecontents of berberine chloride, rutin and baicalin were analyzed by the Analytic HierarchyProcess, and the optimal extraction method was compared with the traditional extractionmethod.The results were as follows:1. The method for simultaneous determination of three kinds of active ingredientsThe high performance liquid chromatography method was determined by differentchromatographic conditions study. The determination was performed on PhenomenexC18(4.6mm×150mm,5μm)，the column temperature was30°C, the mobile phase wasacetonitrile-0.1%H3PO4, the gradient elution was adopted(0â†'25min，B20%â†'40%ï¼›25minâ†'30min，B40%â†'20%). The flow rate was1.0mL·min-1. The detection wavelengthwas265nm. The standard samples and the test samples can be effectively separated underthis method. And the active ingredient contents of berberine chloride, rutin, baicalin can bedetermined very well by this method.2. Optimized ultrasonic extraction method The factors affecting the ultrasonic extraction process such as solvent concentration(70%ethanol,80%ethanol,90%ethanol), ultrasonic power (160W,240W,320W) andextraction time（40min,60min,80min）were optimized by orthogonal test. The optimalultrasonic extraction method was ethanol concentration70%, ultrasonic power240W,extracted three times each60min, and the amount of solvent was10times more than theamount of herbs. The evaluation indexes of the ultrasonic extraction methods are superiorto the evaluation indexes of traditional methods, and the times required in the ultrasonicextraction methods were significantly less than that in the traditional methods too.3. The identification of the compound transdermal patchThe Philippine Violet Herb and Paris polyphylla Smith in the compound transdermalpatch were distinguished by thin-layer chromatography. The same color main fluorescentspots were showed in the chromatograms of the test sample and Herba Violae Philippicae ascontrast drug, while the negative control without fluorescent spots.4. Determining the the active ingredient contents of the compound transdermal patchThe the active ingredient contents of the prepared compound transdermal patch weredetermined by the established high performance liquid chromatography. And the the activeingredient contents of the prepared compound transdermal patch were berberine chloride0.239mg/g, skullcap glycosides2.673mg/g, and rutin1.117mg/g.