1、 Pharmacopoeia standard of Bupleurum chinense
This product is the dried root of Bupleurum chinense dc. or Bupleurum scorzonerifolium willd. According to different characters, they are commonly known as "northern Bupleurum" and "Southern Bupleurum". Excavate in spring and autumn, remove stems, leaves and sediment, and dry.
[character] Bupleurum chinense is cylindrical or conical in shape, 6-15CM long and 0.3-0.8cm in diameter. The root head is swollen, with 3-15 stem bases or short fibrous leaf bases remaining at the top and branches at the bottom. . It is hard and tough, not easy to break, with fibrous section, light brown skin and yellow white wood. The smell is slightly fragrant and the taste is slightly bitter. The root of Bupleurum chinense is thin, conical, with many fine hairy withered leaf fibers at the top, and the lower part is mostly unbranched or slightly branched. The surface is reddish brown or black brown, and there are many fine ring patterns near the root head. The texture is slightly soft, easy to break, the section is slightly flat, and does not appear fibrous. It has lost oil and gas.
[identification] take 0.5g of Bupleurum chinense powder, add 20ml of methanol, sonicate for 10 minutes, filter, and concentrate the filtrate to 5ml as the test solution. Another 0.5g of Bupleurum chinense reference medicinal material was taken, and the reference medicinal material solution was prepared by the same method. . According to the test of thin-layer chromatography (general rule 0502), suck 5 μ l of each of the above three solutions, dot them on the same silica gel G thin-layer plate respectively, develop them with ethyl acetate ethanol water (8:2:1), take them out, dry them, spray them with 2% p-Dimethylaminobenzaldehyde in 40% sulfuric acid, heat them at 60 ℃ until the spots are clear, and inspect them under sunlight and ultraviolet light (365nm). In the chromatogram of the test sample, spots or fluorescent spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material and the chromatogram of the control sample.
[inspection] the moisture content shall not exceed 10.0% (the second method of general rule 0832).
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Acid insoluble ash content shall not exceed 3.0% (general rule 2302).
[extract] according to the hot leaching method (general rule 2201) under the determination method of alcohol soluble extract, the use of ethanol as solvent shall not be less than 11.0%.
[content determination] HPLC method for Bupleurum chinense DC. (general rule 0512)
Assay.
Chromatographic conditions and system suitability test octadecylsilane bonded silica gel was used as filler; Use acetonitrile as mobile phase A and water as mobile phase B to perform gradient elution according to the following table; The detection wavelength was 210nm. The number of theoretical plates should not be less than 10000 according to saikosaponin a peak.
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Time (min) mobile phase a (%) mobile phase B (%)
0～50 25→90 75→10
50～55 90 10
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Preparation of reference solution take an appropriate amount of saikosaponin a and saikosaponin d, weigh accurately, add methanol to make a solution containing 0.4mg of saikosaponin a and 0.5mg of saikosaponin d per 1ml, and shake well.
Preparation of test solution take about 0.5g of powder (passing through No. 4 screen), accurately weigh it, put it into a corked conical flask, add 25ml of methanol solution containing 5% concentrated ammonia test solution, close the stopper, sonicate at 30 ℃ water temperature (power 200W, frequency 40KHz) for 30 minutes, filter it, wash the container and drug residue twice with 20ml of methanol, combine the washing solution with the filtrate, and recover the solvent to dryness. Dissolve the residue with methanol, transfer to a 5ml volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate.
Determination method precisely suck 20 μ l of the reference solution and 10 ～ 20 μ l of the test solution into the liquid chromatograph for determination.
The total amount of saikosaponin a (c42h68o13) and saikosaponin d (c42h68o13) in this product shall not be less than 0.30% according to the dry product.
Decoction pieces
[processing] remove impurities and residual stems from Bupleurum chinense, wash, moisten, cut into thick pieces, and dry.
This product is irregular thick film. The outer skin is dark brown or light brown, with longitudinal wrinkles and root marks. The cut surface is yellowish white and fibrous. Hard quality. The smell is slightly fragrant and the taste is slightly bitter.
[identification], [inspection], [extract], [content determination] are the same as those of Bupleurum chinense.
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This product is shaped like Bupleurum chinense, with a light brown yellow surface, a slight vinegar aroma, and a slight bitter taste.
[extract] according to the hot leaching method under the determination method of alcohol soluble extract (general rule 2201), the use of ethanol as solvent shall not be less than 12.0%.
[identification], [inspection], [content determination] are the same as those of Bupleurum chinense.
Remove impurities from Radix Bupleuri, wash, moisten thoroughly, cut into thick pieces, and dry.
. The outer skin is reddish brown or dark brown. Sometimes there are fine ring lines or fine hairy withered leaf fibers at the root head. The cut surface is yellowish white and flat. It has lost oil and gas.
Vinegar Southern Bupleurum take Southern Bupleurum slices and fry them dry according to the vinegar roasting method (general rule 0213).
This product is shaped like Southern Bupleurum tablets and has a slight vinegar aroma.
[nature, taste and meridian tropism] pungent, bitter and slightly cold. It belongs to the liver, gallbladder and lung meridians.
[functions and indications] evacuate and relieve fever, soothe liver and relieve depression, and elevate Yang Qi. For colds and fever, cold and heat exchanges, chest pain, irregular menstruation, uterine prolapse, anal prolapse.
[usage and dosage] 3 ～ 10g.
[note] the dried rhizome of Bupleurum longiradiatum turcz. has dense links on its surface and is toxic, so it cannot be used as Bupleurum.
[storage] store in a ventilated and dry place to prevent moths.

2、 Chemical constituents of Bupleurum chinense

Bupleurum chinense root contains volatile oil, bupleurum alcohol, oleic acid, linolenic acidpalmitic acid, stearic acid, eicosanoic acid, glucose and saponins, etc. Saikosaponin a、Saikosaponinc、Saikosaponind, saikosapogenin FSaikosaponinE、SaikosaponinG， Longisigenin. It was also reported that saikoside was separated from roots and seeds, which is the general name of various glycosides. In addition, the roots contained α - spinach sterol, Δ 7-stigmasterol, Δ 22 stigmasterolStigmasterol, scutellarin, angelicin. The stems and leaves contain rutin. .
The root of Bupleurum scorzonerifolium contains saponins, fatty oil, volatile oil and Bupleurum alcohol. The stems and leaves contain rutin.
Bupleurum chinense contains rutin, ribitol, 299 ketone, hexanol, α - borlesterol, flavonols, saponins, alkaloids, ascorbic acid, carotene, etc. It can be obtained from flowers, leaves and stems at the flowering and fruiting stagesquercetin 、Isoquercitrin, rutin, isorhamnetin, and isorhamnetin-3-rutinoside.
The root of Bupleurum Folium contains bupleurum, α - spinach sterol, sucrose and polyacetylenes.

3、

1. antipyretic effect
It was proved in early years that a large dose of Bupleurum Decoction (5g crude drug / kg) or alcohol extract (2.5G crude drug / kg) had antipyretic effect on rabbits with artificial fever. The oral decoction or infusion (2 g / kg) also has a mild cooling effect on rabbits with fever caused by typhoid mixed vaccine. Later, it was reported that the antipyretic effect of Bupleurum Decoction was not obvious, while Bupleurum glycoside 200-800 mg / kg orally had a certain reducing normal temperature and antipyretic effect on mice.

2. sedative and analgesic effects
Bupleurum glycosides can sedate mice (pole climbing test) and prolong the sleep of pentobarbital; It has good analgesic effect and strong antitussive effect, but it has no anticonvulsant effect and does not reduce the tension of striated muscle. Some people believe that saikoside can be listed as a central inhibitor.

3. anti inflammatory effect
Bupleurum glycoside orally (600 mg / kg) can significantly reduce dextran and serotonin induced edema in the feet and ankles of rats. In the subcutaneous granulocyst test of rats (croton oil and cotton ball method), it was determined that Chaihu glycoside has the effect of anti exudation and inhibiting the growth of granuloma. Bupleurum is effective either alone or in combination, and its inhibitory effect on granuloma growth is stronger than its anti exudative effect; Quyu Huoxue Formula (Danggui Shaoyao Powder, Taoren Chengqi Decoction, Dahuang Mudanpi decoction, etc.) is opposite to Bupleurum in terms of action intensity, so it is suggested that they should be combined. Saikoside can inhibit the increase of vascular permeability caused by histamine and serotonin, and slightly inhibit pleural effusion; However, carrageenan and acetic acid-induced edema had no protective effect on histamine shock in guinea pigs and anaphylactic shock in mice.

4. anti pathogen effect
It has been reported that Bupleurum injection has a strong inhibitory effect on influenza virus; The unknown oil component distilled from this kind of injection also has a strong inhibitory effect on the virus. It is said to be effective against a certain strain of Mycobacterium tuberculosis. .

5. effects on liver
For the mice with liver dysfunction caused by feeding moldy rice, the elevation of glutamic C and glutamic oxaloacetic transaminase was much lighter than that of the control group without Bupleurum; The effect of saikoside seems to be inferior to that of Bupleurum chinense powder. Oral administration of Bupleurum Decoction (0.5 ～ 1.0 g crude drug / kg) can significantly improve the liver dysfunction caused by typhoid vaccine in rabbits (positive reaction of urobilin); It also has some effects on liver dysfunction caused by alcohol, but it is not as effective as licorice; It is ineffective against organic phosphorus and carbon tetrachloride. It also had no protective effect on experimental liver fibrosis induced by injection of fresh egg yolk solution in rats. The same genus of Bupleurum Xinjiang and Bupleurum rotundifolia are said to have cholagogic effects.

6. cardiovascular effects
Bupleurum alcohol extract can slightly reduce the blood pressure of anesthetized rabbits and inhibit the isolated frog heart, which can not be blocked by atropine. Bupleurum injection has no effect on the heart and blood pressure of eutopic cats with a larger dose. Saikoside can cause a transient hypotensive response and slow down the heart rate in dogs; It also has antihypertensive effect on rabbits, inhibits isolated frog heart, isolated guinea pig atrium, and constricts isolated rabbit ear blood vessels.

7. Other roles
Bupleurum chinense decoction or alcohol extract, given orally to rabbits, can increase blood glucose. The decoction has hemolysis (equivalent to 1/100 of pure saponin prepared by Merk). The saponin content and hemolysis intensity were also different in different places and collection time. Saikoside can prevent stress ulcer in rats, promote the propulsion of small intestine in mice, and enhance the contractile effect of acetylcholine on isolated guinea pig ileum (it cannot enhance the effect of histamine). It also has some excitatory effects on isolated rabbit intestine. . Beichaihu injection has no effect on uterus. The toxicity of Bupleurum is very small. The minimum lethal dose of its alcohol extract for subcutaneous injection in mice is 1.1 ml /10 g (10% aqueous solution). The median lethal dose of Bupleurum glycoside for oral administration in mice is 4.7 g / kg, and the intraperitoneal injection is less than 100 mg / kg. Bupleurum injection has minimal toxicity, and 5 ml / kg intravenous injection has no effect on blood pressure, respiration and heart of cats; 0.2 ml /20 g subcutaneously, non-toxic to mice.
It is said that there is no obvious difference in the effect between domestic Bupleurum and Japanese Bupleurum.
In addition, the flower, leaf and stem extracts of Bupleurum chinense DC. Have cholagogic effects on animals, and also have therapeutic effects on cholecystitis, cholangitis and hepatitis. It can increase the content of cholic acid and bilirubin in bile, and increase the cholesterol bile salt coefficient of bile