| BackgroundLonicera macranthoides Hand.-Mazz is one kind of Lonicera confusa DC. according to the Chinese Pharmacopeia 2010. It is a medicine and food material with a long history. And it has a lot of functions, such as:antibacterial, anti-inflammatory, antivirus, protect liver, antioxidant. The farming area of Xiushan honeysuckle has reached 300,000 mu at present, which has brought tangible benefits to farmers in the locality. To improve its market added value and ability of resisting risk, extend the industrial chain and research deep processing products of Lonicera macranthoides Hand.-Mazz, this article develops a traditional Chinese supplement food-Yin Tian granules, with Lonicera macranthoides Hand.-Mazz as main materials.ObjectiveTo study Yin Tian granules’ extraction technology and preparation technology, investigate its quality and stability, observe its protective effect on experimental liver injury in mice, and lay a good foundation for developing Yin Tian granules into a traditional Chinese supplement food.Methods1. The extract technology:according to the physicochemical properties of effective components in the drug prescription, the co-decoction and water extraction process was selected to extract the effective components. The extraction process was optimized with the extraction yield and the Chlorogenic acid content in extraction as indexes by orthogonal experiments. 2. The forming technology:according to the extracts’properties, the loading agent, the moistening agent and the binder were investigated by single factor experiments. And their dosage was also screened initially. The results of the single factor experiments were optimized by using orthogonal experiment. Then Yin Tian granules were then synthetically investigated and evaluated with taste, particle size, and dissolubility. Finally the best forming technology was screened out.3. Quality standard and Stability test. (1) Identification:TLC method was used to qualitatively identify Lonicera confusa DC., cape jasmine and medical material A. (2) Inspect:according to the preparation principals for IC granules in the appendix I of the primary part of the Chinese pharmacopeia 2010, relative review items were made, including particle size, moisture, dissolubility, and differences among packs. (3) Content determination:the content of chlorogenic acid and geniposide in Yin Tian granules were determined by HPLC. (4) Stability test:through influence factors test and accelerate test, the stability of Yin Tian granules were explored.4. Protection effect on experimental liver injury:(1) Protection effect for acute liver injury caused by CCl4 on mice:Kunming mice were used to do this experiment. They were divided into 6 groups:the normal group, the model group, bifendate group,6.6 g·kg-1.13.2 g·kg-1 and 26.4 g·kg-1 Yin Tian granules groups. After intragastric administration continuously with relative medicine for 14 d, the mice were then administered with 20 mL·kg-1 of olive oil which contains 0.10% CCl4 to cause acute liver injury in mice. And then the activities of ALT and SAT in serum, as well as the content of SOD and MDA in liver were determined. Finally pathological sections with the liver of the mice were made. (2) Protection effect for acute injury caused by alcohol on mice:Kunming mice were used to do this experiment. They were divided into 6 groups:the normal group, the model group, bifendate group,6.6 g-kg-1,13.2 g-kg-1 and 26.4 g·kg-1 Yin Tian granules groups.2 h after intragastric administration with relative medicine, the mice were administered with 50% alcohol except for the normal group. The mice were continuously administered for 14 d. After the last administration with 50% alcohol, the activities of ALT and SAT in serum, as well as the content of SOD and MDA in liver were determined.Results1. The extract technology:the extraction process was determined:taking the components of the prescription Xiushan Lonicera macranthoides Hand.-Mazz., cape jasmine and medical material A, crushing them into coarse powder, adding 15 times water in, and boiling 3 times at the temperature of 70℃, and each time for 1 hour. The extraction yield and the content of chlorogenic acid were 11.18%,59.01% respectively.2. The forming technology:after decompressing and drying the concentrated extract at vacuum, the dry extract was crushed into powder. Then appropriate amount of starch, dextrin. PVP and sweet chrysanthemum sugar was mixed in the extract powder, after which the powder was granulested by wetting method. Dry the wet granules at 60℃for 45 min. And shape particles. Finally seal for packaging.3. Quality standard:(1) Identification:TLC method was used to qualitatively identify the granules and the medical material in the prescription. (2) Inspect:particle size< 15%, and moisture content< 6%. Dissolubility and differences among packs were consistent with the preparation principals for IC granules in the appendix I of the primary part of the Chinese pharmacopeia 2010. (3) Content detection project was accounted by chlorogenic acid, and it should not be lower than 10.00 mg·g-1. (4) Stability test:through the influence factor test investigation, we draw a conclusion that the Yin Tian granules should avoid long-time exposure with highlight when produced, packed and stored.4. Protection effect on experimental liver injury:(1) Protection effect for acute liver injury caused by CCl4 on mice:compared with the normal group, the activity of AST and ALT in serum of the model group was obviously increased, and the content of MDA in liver was also increased, while the activity of SOD in liver was decreased, all of which had great difference (P<0.01). It indicated the model was successful. Compared with the model group, the activities of AST and ALT in serum of the 6.6 g-kg-1,13.2 g-kg-1 and 26.4 g-kg-1 Yin Tian granules groups were obviously decreased (P<0.01). and the content of MDA in liver of 13.2 g-kg-1 and 26.4 g·kg-1 Yin Tian granules groups was obviously decreased (P<0.01), while the activity of SOD in liver of 13.2 g·kg"’and 26.4 g·kg" Yin Tian granules groups was obviously increased (P<0.01). The result indicated that Yin Tian granules have protection effect on experimental liver injury caused by CCl4 on mice. (2) Protection effect for acute injury caused by alcohol in mice: compared with the normal group, the activities of AST and ALT in serum of the model group were obviously increased, and the content of MDA in liver was also increased, while the activity of SOD in liver was decreased, all of which had great difference (P< 0.01). It indicated the model was successful. Compared with the model group, the activity of AST in serum of the 6.6 g-kg-1.13.2 g-kg-1 and 26.4 g-kg-1 Yin Tian granules groups was greatly decreased (P<0.01); the activity of ALT in serum of the 6.6 g·kg-1 13.2 g-kg-1 and 26.4 g-kg-1 Yin Tian granules groups was obviously decreased (P<0.05 or P<0.01); the activity of SOD in liver of 13.2 g-kg-1 and 26.4 g-kg-1 Yin Tian granules groups was obviously increased (P<0.01); the content of MDA in liver of 6.6 g-kg-1, 13.2 g-kg-1 and 26.4 g-kg-1 Yin Tian granules groups was obviously decreased (P<0.05 or P<0.01). The result indicated that Yin Tian granules have protection effect on experimental liver injury caused by alcohol on mice.ConclusionYin Tian granules were obtained by follow steps:taking appropriate amount of Xiushan Lonicera macranthoides Hand.-Mazz., cape jasmine and medical material A according to the prescription; crushing them into coarse powder, and adding 15 times amount of water in; boiling 3 times at the temperature of 70℃, and each time for 1 hour; concentrating the extract; drying and crushing the extract; mixing 5 times amount of loading agent and 5% PVP into the extract; granulating the powder by wetting method with 60% alcohol; drying the wet granules and sifting. The Yin Tian granules prepared as this technology have controllable quality and good stability. And it also has protection effect on experimental liver injury caused by CCl4 and alcohol on mice. |
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