Analysis Of The Active Constituents And The Studies Of Relieving Asthenopia In Dendrobe Medlar Tablet

The people suffering from eye diseases have on the acceleration in recent years. There are several reasons such as spending long time on the TV and computer, oppositely shortening the rest time of eye. The chronic diseases such as diabetes, hypertension can cause the increasing incident of cataracts,glaucoma, etc. Dendrobe Medlar Tablet is a healthy food which the mind active ingredient are lutein, beta carotene and procyanidins. The topic has been studied the detection method and the determination the content of active ingredient, and it researched the safety experiments and the test of relieving asthenopia. This study provides the basis for both Dendrobe Medlar Tablet’s quality control and the study of the other relieving asthenopia tablet. The research mainly includes two aspects.PartⅠAnalysis of the active constituents in Dendrobe Medlar Tablet1To establish the method that determinated the content of lutein and beta carrots simultaneously in Dendrobe Medlar Tablet by a new HPLC Method of Substitute for Reference Substance.Methods: Ultrasonic hydrolysis3min in60℃, and ultrasonic extraction by the solvent of acetonitrile with0.1%BHT. constant volume. chromatographic column：Thermo Beta Basic-18Column(250mm×4.6mm，5μm). Mobile phase: composed by A Methanol、B water、C THF Tetrahydrofuran，gradient elution; Flow rate：1.0ml/min; column temperature:30℃; UV detection wavelength:474nm. Result: the average content of lutein is11.52mg/g, and average standard addition recovery rate of lutein is99.55%, relative standard deviation was2.47%. the average content of beta carrots is3.49mg/g, average standard addition recovery rate of beta carrots is98.92%, relative standard deviation was4.69%2The detection of lutein and the analysis of ionization mechanism by HPLC-MS Method: acetonitrile extraction solvent is acetonitrile, ultrasonic extraction: chromatographic column：Thermo Beta Basic-18Column(150mm×2.1mm，5μm); Flow rate:0.2ml; Mobile phase: methanol-water with0.8%formic acid(95:5); Column temperature:30℃; ion source: ESI(+);scan patterns MS3; the full-scan mass spectrum with in-source dissociation; Multistage scanning used fragment ions scanning.Result: the experiment using the mode of source within the decomposition-full scan mode lost toluene characteristic fragment ions475.2[M-C7H8]+,scanned for secondary level, get the molecular ion of mass-to-charge ratio457.2,439.3,362.4. providing a new method for lutein qualitative and data3The study of content determination in Dendrobe Medlar Tablet by Vanillin-hydrochloric acid methodMethod: extracting solution was60%methyl alcohol, the time of ultrasonic treatment is45min by the temperature of40℃; adding6ml Vanillin methanol with the concentration of4%, and3ml hydrochloric acid reacted30min lucifugely, then we could detect absorbance in wavelength of500nm with the methanol as blank control. Result: average standard addition recovery rate of procyanidine by Vanillin-hydrochloric acid method is99.8%, relative standard deviation was2.86%.4To develop a new method using HPLC for determination content of procyanidine in Dendrobe Medlar Tablet.Method: Extracting solution was methyl alcohol and the time of ultrasonic treatment is45min. Chromatographic column：Ultimate ColumnTM C18（4.6mm×250mm，5μm）; UV detection wavelength:280nm. Result: average standard addition recovery rate of procyanidine by HPLC is98.9%, relative standard deviation was3.4%. PartⅡ The study of safety experiments and relieving asthenopia in Dendrobe Medlar Tablet1The safety experiments of Dendrobe Medlar Tablet Method: the study included: thirty-day feeding, acute toxicity test, Ames test, Bone marrow of mice addicted to dye red blood cells micronucleus test and Sperm aberration test in mice showed the product non-toxic.2The test of relieving asthenopia in Dendrobe Medlar Tablet Method: A total of120volunteer were divided into test group took and placebo group, test group took Dendrobe Medlar Tablet and placebo group took placebo. Two pieces at a time,three times a day,60d.Result: comparing with test group and placebo group, symptoms improved and the symptoms of total points had significant different(P<0.05). visuognosis persistence had significant different and increased more than10%comparing between test group and placebo group. We can judged that the product can relieve asthenopia.