1、1Research on Kang Fu Yan Granule Technology and Quality StandardAbstract. This paper aims to confirm the preparation technology of Kang Fu Yan granule and make its quality standard. Orthogonal experimental method is applied to investigate the alcohol extraction volume of rhizoma corydalis (yanhusuo)
2、, the volatile oil extraction of rhizoma atractylodis, rhizoma cyperi, angelica and rhizome chuanxiong, the decoction dregs of rhizoma atractylodis and other medicinal materials, and the six-taste water extraction technology of dandelion and other medicinal materials, hence optimizing the technologi
3、cal parameters. The best technological preparation for Kang Fu Yan granule is confirmed in the technological researches. This paper concludes that the method and indexes in this experiment are applicable to the preparation technology of Kang Fu Yan granules. Key words: Kang Fu Yan granules, preparat
4、ion technology 1. Instruments and Reagents Instruments: LC2050 high performance liquid chromatograph, LC2400 ultraviolet detector, N2000 chromatogram workstation Reagents: paeoniflorin reference substance, rhizoma 2atractylodis reference medicinal materials, angelica reference medicinal materials, r
5、hizome chuanxiong reference medicinal materials,-cyperone reference substance, and yanhusuo reference medicinal materials; acetonitrile: chromatographic purity; redistilled water (made by ourselves); analytical reagent 2. Technological Researches 1 (1) Investigate the alcohol extraction volume of ya
6、nhusuo; (2) Investigate the volatile oil and extraction time of rhizoma atractylodis, rhizoma cyperi, and angelica and rhizome chuanxiong by steam distillation; (3) Investigate the decoction dregs after volatile oil extraction, the dandelion six-taste water extraction technology and water volume. 2.
7、1 Alcohol Volume Experimental Methods Weigh three pieces of 240g yanhusuo, pour 70% of alcohol based on the designed alcohol volume, extract them respectively at twice (1.5 hours for each time), combine the extraction liquid and filter it. The filtered liquid recovers the alcohol until no alcohol od
8、or; concentrate the alcohol until becoming thick paste, and dry it until becoming constant weight; weigh it respectively and calculate the receiving rate of dried paste. 3The experimental results show that the extraction percentages of 12 times alcohol and 10 time alcohol are close to those of 10 ti
9、mes alcohol and 8 times alcohol, the 12 times alcohol and 10 time alcohol have the best effect, 8 times alcohol and 6 times alcohol have the poor effect. Therefore, it is the most economical to select 10 times alcohol and 8 times alcohol from the perspectives of production cost reduction and energy
10、saving. 2.2 Investigation on the Volatile Oil Extraction Technology of Rhizoma Atractylodis, Rhizoma Cyperi, Angelica and Rhizoma Chuanxiong and other Medicinal Materials 2.2.1 Weigh 1/2 amount of medicinal materials required in the prescription for three times respectively: 114g rhizoma atractylodi
11、s, 114g rhizoma cyperi, 180g angelica, 144g rhizoma chuanxiong, and mark them A,B,C respectively. Extract volatile oil with steam distillation method respectively, and record the volatile oil amount when extracting for 2 hours, 4 hours, 5 hours, 6 hours, 7 hours and 10 hours. The results indicate th
12、at the volatile oil is almost extracted completely when extracted for 6 hours, so the extraction time is confirmed to be 6 hours for the sake of energy saving. 2.2.2 Researches on the Decoction Dregs of Rhizoma 4Atractylodis and Other Medicinal Materials and the Six-taste Water Extraction Technology
13、 of Dandelion and Other Medicinal Materials Weigh 1/2 amount of medicinal materials required in the prescription for three times respectively: 114g rhizoma atractylodis, 114g rhizoma cyperi, 180g angelica, 144g rhizoma chuanxiong. Extract volatile oil by steam distillation method respectively, colle
14、ct aqueous solution and decoction dregs and set aside. Also, weigh 180g dandelion, 180g patrinia, 180g semen coicis, 144g red paeonia, and 108g alisma orientale for three times respectively. Combine decoction dregs with the three pieces of dandelion and other medicinal materials respectively, and po
15、ur water to decoct them respectively at twice (1.5 hours for each time), and then filter them respectively and combine the filtered liquid, pour the liquid filtered after the volatile oil extraction respectively, decompress and concentrate the mixture until the relative density becomes 1.201.15 (50)
16、 extract, pour alcohol until the alcohol content reaches 50%, cool for 24 hours and filter, decompress the filtered liquid and recover the alcohol until the relative density is concentrated to be 1.301.35 (50) thick paste, and decompress it until being dried and constant 5weight. The experimental re
17、sults show that the percentages of 12 times alcohol and 10 times alcohol are close to those of 10 times alcohol and 8 times alcohol, 12 times alcohol and 10 times alcohol have the best extraction effect, 8 times alcohol and 6 times alcohol have the poor extraction effect. Therefore, it is the most e
18、conomical to select 10 times alcohol and 8 times alcohol from the perspectives of energy saving and production cost reduction. 2.3 Preparation Technological Process Take 11 medicinal materials in the prescription; 70% of alcohol is used for yanhusuo and extract it twice with reflux extraction method
19、 (1.5 hours for each time), pour 70% of alcohol which are 10 times and 8 times of all the poured medicinal materials, filter, and use the filtered liquid to recover alcohol until the relative density is concentrated to be 1.13 (50) extract, and set aside. Extract the volatile oil of the rhizoma atra
20、ctylodis, rhizoma cyperi, and angelica and rhizoma chuanxiong with steam distillation method for 6 hours. Pour water into the decoction dregs, dandelion and other six medicinal materials and decoct them twice, combine the filtered liquid, pour the liquid filtered after the volatile oil extraction, d
21、ecompress it until the relative density is 6concentrated to be 1.10-1.15 (50) extract, pour alcohol until its volume reaches 50%, cool 24 hours, filter, and decompress the filtered liquid to recover the alcohol, and pour the above extract, and concentrate it until the relative density is 1.301.35 (5
22、0) thick paste, and then pour a proper amount of dextrine and stevia rebaudianum, mix evenly, make it into granules and dry, pour the above volatile oil, and pack finally (1000g). 3. Researches on Quality Standard 3.1 Thin-layer Chromatography Identifications 3.1.1 Rhizoma Atractylodis1 Take 20g sam
23、ple, porphyrize it, pour 50ml diethyl ether, and process for 20 minutes with ultrasonic, volatilize the diethyl ether with the filtered liquid; pour 1ml absolute ethyl alcohol into the residue to let it dissolved; take 1g rhizoma atractylodis reference medicinal material and make it into solution wi
24、th the same method. According to the thin-layer chromatography experiment, absorb the above 10ul solution respectively, and drop them respectively onto the silica-gel G thin board that takes sodium carboxymethyl cellulose as adhesive; use petroleum ether (6090) and acetic ether (20:1) as the develop
25、ing solvent, unfold, take out and dry it, spurt 710% of vitriol and alcohol solution, and bake at 105 until the color of spots is clear. In the reference solution chromatography, the chromatography position corresponding to the reference medicinal materials shows the same color spots (See figure 1).
26、 3.1.2 Angelica and Rhizome Chuanxiong2 Take 0.5g Angelica and rhizome chuanxiong reference medicinal materials respectively, pour 20ml diethyl ether into them respectively, and process for 20 minutes with ultrasonic, volatilize the diethyl ether with the filtered liquid, pour 1ml absolute ethyl alc
27、ohol into the residue to let it dissolved as the reference medicinal material solution. Also, based the proportions in prescription and technological process, prepare the negative reference samples without Angelica and rhizome chuanxiong, and make it into negative reference solution with the same me
28、thod. Based on the thin-layer chromatography experiment, absorb the above reference medicinal material solution and the 10ul reference solutions respectively under the identification (1), drop them respectively onto the same silica-gel G thin board, use normal hexane and acetic ether (9:1) as the de
29、veloping solvent, and place it under the UV-light (365nm) to be inspected. In the reference solution 8chromatography, the chromatography position corresponding to the reference medicinal materials shows the same color spots. However, the negative reference solution has no spots on the corresponding
30、positions, which is negative with no interference (See figure 2). 3.1.3 -Cyperone 3 Take -Cyperone reference substance and pour absolute ethyl alcohol to be solution (1ml/1mg) as the reference solution. Based on the thin-layer chromatography experiment, absorb the reference solutions respectively un
31、der the identification (1) and 10ul above reference solutions respectively, drop them respectively onto the same silica-gel G thin board, use petroleum ether and acetic ether (17:3) as the developing solvent, unfold, take out and dry it, spurt dinitrophenyl hydrazine ethanol solution, and lay it asi
32、de until the color of spots is clear. In the reference solution chromatography, the chromatography position corresponding to the references shows the same color spots (see figure 3.) 3.1.4 Yanhusuo4 Take 20g sample, porphyrize it, pour 50ml methanol, process for 30 minutes with ultrasonic, filter an
33、d dry by distillation, pour water into residue to dissolve it, add stronger ammonia solution until being alkaline, extract it with diethyl ether 9for three times, combine diethyl ether liquid, and pour 1ml methanol into residue until to be dissolved as the reference solution. Also, take 0.1g yanhusu
34、o reference medicinal material, and make it into reference medicinal material solution with the same method. Based on the thin-layer chromatography experiment, absorb 5ul above reference medicinal material solutions respectively, and drop them respectively onto the same silica-gel G thin board that
35、uses 1% sodium hydroxide solution as adhesive, use normal hexane, chloroform and methanol (7.5:4:1) as the developing solvent, and place it in the jar with saturated developing solvent, unfold, take out and dry it, fumigate it with iodine steams until the color of spots is clean, and inspect it in t
36、he sun. In the reference solution chromatography, the chromatography position corresponding to the reference medicinal materials shows the same color spots. After the iodine is volatilized completely in the air, put it under the UV-light (365nm) to be inspected. In the reference solution chromatogra
37、phy, the chromatography position corresponding to the reference medicinal materials shows the same color spots with fluorescent light (see figure 4). 3. 2 Inspections 5 3. 2.1 Granularity 10Take 5 bags of samples from 20070506, 20070507 and 20070508 respectively, pour them out and weigh them respect
38、ively, and sift them in the prescription sieves. Keep the sieves horizontal when sifting the sample, and slightly sift from left to right for 3 minutes; and the total amount of the granules and powder that do not pass sieve 1 but can pass sieve 4 (TAGP) cannot exceed 8.0%. The results are as shown i
39、n table 1. The result indicates that the granularity of the three groups of samples all can be less than 8%. 3. 2.2 Moisture Content Based on the H moisture content first inspection method in the appendix IX of volume 1 of 2005 Pharmacopoeia of Peoples Republic of China, the result is as shown in ta
40、ble 2. The result indicates that the moisture Content of the three groups of samples all can be less than 5.0%. 3. 2.3 Dissolubility Take 10g samples from 20070506, 20070507 and 20070508 respectively, pour hot water into them and mix for 5 minutes, observe them immediately. As a result, the three groups of samples all can be totally dissolved and have no burnt crumbs and other sundries. 3. 2.4 Content Uniformity
