Research On Preparative Technology And Quality Standard Of Rhubarb Dispensing Granule

1 ObjectiveChinese medicinal granule, as a supplement of Chinese traditional medication decoction pieces, was a new dosage form with more convenient, more scientific, healthier and safer characteristics. To its craft's improvement, it may effectively enhance the use of the traditional Chinese medicine resources, guarantees or even enhances efficacy of Chinese medicinal granule. At present, because the extraction method of Chinese medicinal granule is different from before, and the effective components between Chinese medicinal granule and Chinese traditional medication decoction pieces have differences. What's more, in each kind of Chinese herb includes many kinds of known or unknown ingredient, activated or passive ingredient, and the action mechanism was still not clear. They cause difficulty to establish quality standard.Researching from the technologies of extraction, purification, concentration and desiccation of rhubarb dispensing granule, we could find out the alternative rules of anthraquinones in the production process and thus establish the whole set technique of production from material to dispensing granule. Referring to the test methods of rhubarb listing in the first part of the Chinese pharmacopoeia 2005, it makes some improvements to rhubarb dispensing granule's quality analysis method, and establishes appropriate quality rating system of it. 2 Methods2.1 Material selection for rhubarb dispensing granuleAccording to the first part of the Chinese pharmacopoeia 2005, we take some tests to the rhubarb from Sichuan, Gansu and Tibet.2.2 Extraction process research of rhubarb dispensing granuleAs the anthraquinones content to be evaluation indicator, we first adopt L9(34) Orthogonal Design to select out the best water extraction. Second, we take single factor exploration to find out the suitable temperature, material particle size, and alcohol concentration.Third,we adopt L9(34) Orthogonal Design to select out the best alcohol extraction. Finally, comparing to the different time of extraction of alcohol and water, we could find out the influence factor of anthraquinones' dissolves.2.3 Separation and purification process research of extracted liquids of rhubarb dispensing granuleTaking the adsorption and desorption as evaluation indexes, we compare the performances of adsorption and desorption of six types of macroporous resin to find out the best type macroporous resin to separate the anthraquinones in Rheum.And then continue to study the conditions and parameters of purifying total Anthraquinones in rhubarb extracted liquid with the best resin.2.4 Desiccation research of purification liquids rhubarb dispensing granuleTakes the Chinese rhubarb concentrated solution to carry on the freeze-drying, the vacuum drying and the spray drying experiment separately. As the total anthraquinones content to be evaluation indicator, and combines with factors of water-solubility, the outward appearance, absorption of moisture in the air for three hours of the powder to find out the best Desiccation method.2.5 Study on establishing quality standard of rhubarb dispensing granuleReferring to the test methods of rhubarb listing in the part of the Chinese pharmacopoeia 2005, it makes some improvements to rhubarb dispensing granule's quality analysis method, and establish appropriate quality standard of it. Rhubarb dispensing granules which are made from different materials of different companies have been tested by TLC, HPLC and Fingerprint atlas. 3 Result3.1 Material selection for rhubarb dispensing granuleThe results indicate that rhubarb from Tibet does not meet the pharmacopoeia requirement because of containing rhapontin. The quantity of 5 kinds of the anthraquinones in rhubarb from Sichuan is higher than from Gansu, accounting for 2.85% of total material quantity.3.2 Extraction process research of rhubarb dispensing granuleThe optimal extraction conditions were obtained as 60 mesh material powder adding 12-fold times of 60 percent alcohol, no immersion,percolating 0.5 hours in 90℃extracting three times.3.3 Separation and purification process research of rhubarb extracted liquidsThe result shows that the separation effects are different with different type's macroporous resin. The DM130 resin is the suitable material. On the basis of above, we continue to study the conditions and parameters of purifying total Anthraquinones in rhubarb extracted liquid with DM130 resin. When the concentration is 0.2g/mL, the pH is about 6 to 7 and the flow rate is 20d/min, the adsorption condition are appropriate; when the alcohol concentration is 60%, the effect of desorption is satisfactory, with the dosage is 17 times column volume.3.4 Desiccation research of rhubarb purification liquidsThe result shows that spray drying technology is suitable for rhubarb purification liquids'drying. To continue studying the conditions of spray drying,we knows that when the air humidity is 50% to 60%,Inlet temperature is 180℃,Outlet temperature is 110℃,it has good drying efficiency.3.5 Study on quality standard of rhubarb dispensing granule3.5.1 Thin layer ChromatographicThe optimized results are as follow:The better developing solvent in the to isolate the anthraquinones of rhubarb dispensing granule is cyclohexane-ether-ethyl acetate-formic acid (20:5:2:1) supernatant. Carring 7 raid of rhubarb dispensing granule on the thin layer chromatograph distinction, under the 365nm uviol lamp to inspect, we found that for tries in the chromatograph, in with the comparison raw material for medicine chromatograph relevant position, assumes the same five orange yellow fluorescence spots; In and comparison chromatograph corresponding position, obviously same orange yellow fluorescence spot.3.5.2 High Performance Liquid ChromatographyThe condition is:Chromatographic stationary phase:ODSMobile phase:methyl alcohol-0.1% phosphoric acid (85:15)Flow rate:1.0mL/minColumn temperature:25℃Wavelength:254nmUnder these conditions, appearance time of Aloeemodin, Rhein, Emodin, Chrysophanol, Physcion are 4.48min,5.46min,7.09min,8.03min, and 9.00min.Examine probational liquid's preparation:Takes the rhubarb powder approximately 0.1g joins methyl alcohol 25mL precisely, the water bath backflow withdraws for 1 hour, has filtered, takes continues filtrates 5mL, the evaporation, joins 2.5mol/L sulfuric acid 10mL, the supersonic 5 minutes, joins the chloroform lOmL heating backflow for 1 hour, cools to the room temperature, pours into the tap funnel to plant, the vessel with the few chloroform wash, merges the tap funnel to plant, divides takes the chloroform level, the acid liquor level washes with the chloroform three times, each time lOmL, the merge chloroform fluid, the reduced pressure recycling to does, adds the methyl alcohol to dissolve and to shift to the 10mL measuring flask, adds the methyl alcohol to dilute to the scale division.Under these conditions, we could obtain the sum of the content of Aloeemodin, Rhein, Emodin, Chrysophanol, Physcion from No.1 to No.7 rhubarb dispensing granule are 17.44 mg/g,10.21mg/g,14.84mg/g,14.20mg/g,17.18mg/g,15.17mg/g.21.49mg/g.3.5.3 Research on Fingerprint of rhubarb dispensing granulesThe condition is:Chromatographic stationary phase:ODSMobile phase:A is Methyl cyanide, B is 0.1%phosphoric acid. The gradient elution process are from 0～30 min, mobile phase A from 25% to 60%, mobile phase B from 75% to 40%; from 30～50 min, mobile phase A from 60% to 70%,mobile phase B from 40% to 30%; from 50-60min, mobile phase A from 70% to 85%,mobile phase B from 30% to 15%; from 60-70min, mobile phase A from 85% to 100%.mobile phase B from 15%～0 %.Flow rate:1.0mL/min Column temperature:25℃Wavelength:254nmCarring on the similarity appraisal of 1-7 raids of rhubarb dispensing granule, altogether has the peak is 33, except the batch number 7 rhubarb dispensing granule's chromatograph peak similarity is 69.38%, other various peaks similarity achieves above 80%.And then carring on the similarity appraisal except the batch number 7 to other 6 chromatograph peak, various peaks similarity achieve above 90%.Rhubarb dispensing granules which are made from different materials of different companies have been tested by TLC, HPLC and Fingerprint atlas.The result shows that rhubarb dispensing granules which are made from the materials 1(090302) would be better.4 ConclusionThe present study use the content of Anthraquinones as a target to search the suitable technique of production which with the procedure more stable and more controllable. And research on the quality standard to establish a quality rating system with TLC,HPLC and Fingerprint atlas. This will give a reference to the rhubarb dispensing granules of its quality control and quality detection.