1、 Rubia cordata pharmacopoeia standard
This product is the dried roots and rhizomes of Rubia cordifolia L. Excavate in spring and autumn to remove sediment and dry.
[character] the rhizome of this product is nodular, with clumps of roots of different thickness. The root is cylindrical, slightly curved, 10-25cm long, 0.2-1cm in diameter; the surface is reddish brown or dark brown, with fine longitudinal wrinkles and a few fine root marks; . It is brittle, easy to break, with flat section, narrow skin, purplish red, broad wood, light yellow red, and most conduit holes. The breath is slight, the taste is slightly bitter, and the tongue is pricked by chewing for a long time.
[identification] (1) cross section of the root of this product: 6-12 rows of cork cells, containing brown matter. Some parenchymal cells in the inner layer of the thrombus contained reddish brown granules. Phloem cells were smaller. The cambium is not very obvious. . Parenchyma cells contain calcium oxalate needle crystal bundles.
(2) ; The ether layer is colorless, and it shows sky blue fluorescence when observed under ultraviolet light (365nm).
(3) Take 0.5g of this product powder, add 10ml of methanol, sonicate for 30 minutes, filter, and concentrate the filtrate to 1ml as the test solution. Another 0.5g of Rubia control medicinal material was taken, and the control medicinal material solution was prepared by the same method. Then take the control sample of alizarin and add methanol to make a solution containing 2.5mg per 1ml as the control solution. . In the chromatogram of the test sample, fluorescent spots of the same color appear at the positions corresponding to the chromatogram of the control medicinal material and the chromatogram of the control sample.
[inspection] the moisture content shall not exceed 12.0% (the second method of general rule 0832).
The total ash content shall not exceed 15.0% (general rule 2302).
Acid insoluble ash content shall not exceed 5.0% (general rule 2302).
[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 9.0%.
[content determination] determine according to high performance liquid chromatography (general rule 0512).
Chromatographic conditions and system suitability test octadecylsilane bonded silica gel was used as filler; The mobile phase was methanol acetonitrile-0.2% phosphoric acid solution (25:50:25); The detection wavelength is 250nm. The number of theoretical plates should not be less than 4000 according to the peak of alizarin and hydroxyalizarin.
Preparation of reference solution take a proper amount of Alizarin reference substance and hydroxyalizarin reference substance, weigh accurately, and add methanol to make a solution containing 0.1mg of alizarin and 40 μ g of hydroxyalizarin per 1ml.
Preparation of test solution take about 0.5g of powder (passing through No. 2 screen), weigh accurately, place in a corked conical flask, add 100ml of methanol precisely, close the stopper, weigh, place overnight, sonicate (power 250W, frequency 40KHz) for 30 minutes, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 50ml of continuous filtrate, evaporate to dryness, add 20ml of methanol-25% hydrochloric acid (4:1) mixed solution to dissolve the residue, heat and hydrolyze in a water bath for 30 minutes, immediately cool, add 3ml of triethylamine, mix well, transfer to a 25ml volumetric flask, add methyl alcohol to the scale, shake well, filter, and take the continuous filtrate.
The determination method is to precisely suck 10 μ l of the reference solution and 20 μ l of the test solution, inject them into the liquid chromatograph, and determine.
Calculated as a dry product, the content of Alizarin (c17h15o4) shall not be less than 0.40%, and that of hydroxyalizarin (c14h8o5) shall not be less than 0.10%.
Decoction pieces
[processing] alizarin is used to remove impurities, wash, moisten, cut into thick pieces or sections, and dry. This product is irregular thick piece or segment. The root is cylindrical, with reddish brown or dark brown outer skin, with fine longitudinal lines; The peeling part of the skin is yellowish red. The cut skin is narrow, purplish red, the wood is broad, light yellow red, and there are many conduit holes. The breath is slight, the taste is slightly bitter, and the tongue is pricked by chewing for a long time.
[content determination] the content of Alizarin (c17h15o4) shall not be less than 0.20% and that of hydroxyalizarin (c14h8o5) shall not be less than 0.080%.
[identification], [inspection], [extract] are the same as that of medicinal materials. Alizarin charcoal take the tablets or segments of alizarin and stir fry them according to the method of stir frying charcoal (general rule 0213) until the surface is burnt black.
This product is shaped like a piece or segment of sycamore, with dark brown surface and brown interior. It tastes bitter and astringent.
[identification] take 0.4g of this product powder, add 5ml of diethyl ether, shake for several minutes, filter, add 1ml of sodium hydroxide test solution to the filtrate, shake, stand still to make it layered, the water layer is red, the ether layer is colorless, observe under the UV lamp (365nm), it shows sky blue fluorescence.
[inspection] the moisture content is the same as that of medicinal materials, and shall not exceed 8.0%.
[extract] the same as that of medicinal materials, not less than 10.0%.
[nature, taste and meridian tropism] bitter, cold. .
[functions and indications] cool blood, remove blood stasis, stop bleeding, and dredge meridians. It is used for hematemesis, bleeding, metrorrhagia, trauma bleeding, blood stasis, menstrual closure, arthralgia, tumescent pain.
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[storage] put it in a dry place.

2、 Chemical constituents of Rubia cordifolia

1. Qian Cao root contains anthraquinone derivatives: alizarin,Hydroxyalizarin, isoalizarin, 1-hydroxy-2-methylanthraquinone, 1,4 dihydroxy-6-methylanthraquinone, norrosaldehyde,Emodin methyl ether, 2-dihydroxyanthraquinone-2-o - β - D-xylose (1 → 6) - β - D-glucoside, 1,3,6-trihydroxy-2-methylanthraquinone-3-0 - (6 '- O-acetyl) - α - rhamnose - (1 → 2) - β - glucoside, 1,3,6-trihydroxy-methylanthraquinone-3-o - α - rhamnol (1 → 2) - β - glucoside, 1,3-dihydroxy-2-hydroxymethylanthraquinone-3-o-xylose (1 → 6) - glucoside, 1-acetoxy-6-hydroxy-2-methylanthraquinone-3 -o - α - rhamnose (1 → 4) - α - glucoside, 1,3,6-trihydroxy-2-methylanthraquinone-3-o - (6' - O-acetyl) - β - D-glucoside, 1,3,6 trihydroxy-2-methylanthraquinone-3-o - (3 '- acetyl) - α - rhamnose (1 → 2), glucoside and isoalizarin-3-o - β - D-glucoside.

2. naphthoquinone derivatives: alizarin, 2-carbamoyl-3-methoxy-1,4-naphthoquinone, 2-carbamoyl-3-hydroxy-1,4-naphthoquinone, dehydro - α - lapachone, furan alizarin, dihydro alizarin, alizarin lactone, 2'- methoxy alizarin, 2'- hydroxy alizarin, 1-methoxy-2'- hydroxydihydro alizarin, 1', 2'- dihydroxydihydro alizarin, 2-fusimethyl-3-isopentenyl-2, 3-epoxy-1,4-naphthoquinone, alizarin polyphthaloquinone.

3. naphthalene hydroquinone derivatives: 2-methyl-3-isopentenyl-1,4-naphthalene hydroquinone-bis - β - D-glucoside, 3-methyl-2 - (3'- hydroxy) - isopentyl-1,4-naphthalene hydroquinone-1-0 - β - D-glucoside, etc.

4. triterpenoids: Rubia nigra terpene aRubia nigra terpeneB， Alizarin apol D, oleanolic acid acetate, oleanolic aldehyde acetate; Others: 6-methoxyduchenne glycosidic acid,Scopoletin Pavilion, fatty acids,β - sitosterolandCaroteneEt al.

5. it is also reported that Rubia still contains 1.3,6-trihydroxy-2-methylanthraquinone-3-0-xylose (1 → 2) - (6 '- 0-acetyl) - glucoside, Rubia terpene triol, and d-isolarch alcohol.

3、 Pharmacological action of Rubia cordifolia

1. hemostatic effect: rabbits can obviously promote blood coagulation within 2-4 hours after oral administration of appropriate amount of Rubia warm extract, or 30-60 minutes after intraperitoneal injection of the same dose of Rubia solution. The calcium recovery time, prothrombin time and thromboplastin time on the white pottery were shortened. Rubia charcoal can also significantly shorten the tail bleeding time of mice. . After oral administration of warm extract, rabbits can partially correct the prolongation of recalcification time caused by heparin and partial thromboplastin time of white clay in vivo. The blood calcium recovery time and partial thromboplastin time of white clay in normal rabbits were 110 seconds and 45 seconds, respectively; After 60 minutes of heparin injection, the time of heparin injection was more than 600 seconds; After perfusing Rubia warm extract (100%50ml) for 90mm, and then injecting heparin for 60 minutes, it was 180 seconds and 64 seconds, respectively. Therefore, Rubia can be used to treat bleeding diseases caused by excessive heparin or heparin like substances. Rubia and protamine were experimentally studied and proved. . The effect of protamine, toluidine blue and Rubia officinalis on the anticoagulant effect of Salvia miltiorrhiza results: the clotting time of thrombin in normal control was 9.5 seconds, 12.5% salvianized plasma was 50.8 seconds, 1% protamine shortened 34.8 seconds, while Rubia officinalis prolonged 1.2 seconds. It is suggested that the procoagulant effect of Rubia cordifolia has its particularity.

2. antiplatelet aggregation: in vitro, alizarin has a strong inhibitory effect on rabbit platelet aggregation induced by arachidonic acid (AA) and collagen, with an IC50 of about 86.6mmol / L. it also has a certain inhibitory effect on platelet activating factor (PAF) - induced aggregation. 2-methyl-1,3,6-trihydroxyanthraquinone has a strong inhibitory effect on collagen induced platelet aggregation and thrombin. Although the effects induced by AA and PAF were weak, they were still significant. 1, 3, 6. trihydroxy-2-methylanthraquinone-3-0 - (3 '- O-acetyl) - 2-rhamnose (1 → 2) - glucoside, isoalizarin-3-0 - β - D-glucoside and isoalizarin are selective inhibitors of collagen induced platelet aggregation.

3. leukocyte elevating effect: the crude extract of Rubia has leukocyte elevating effect. One of its effective ingredients for increasing leukocytes is phenolic carboxylic glycosides with aromatic rings. The derivative of this effective ingredient, alizarin diester, has been synthesized artificially. Alizarin diester had an increasing effect on white blood cells of normal mice and large mice. Mice: 2.5mg/mouse orally, white blood cells increased significantly after 8 hours of administration, and the administration group was 151.9% of the control group (P < 0.05). . Alizarin diester can also promote the proliferation and differentiation of bone marrow hematopoietic stem cells in mice and prevent cyclophosphamide induced leukopenia in dogs.

4. antitussive and expectorant effects: ammonia spray cough inducing method and phenol red excretion method, respectively, proved that oral administration of qiancaohen Decoction 75g/kg to mice had obvious antitussive and expectorant effects, but the filtrate precipitated with ethanol was ineffective.

5. antibacterial effect: the in vitro test showed that the water extract of Rubia Radix has certain inhibitory effect on Staphylococcus aureus, Streptococcus pneumoniae, influenza bacilli and some skin fungi. Alizarin also has inhibitory effect on Staphylococcus aureus.

6. anticancer effect: RA (rubiaakane) -v10mg/kg, ra- Ⅶ 4mg/kg, a cyclohexpeptide compound contained in the root of Qian Cao, was injected intraperitoneally for 5 consecutive days, which had significant curative effect on mouse lymphocytic leukemia P388. Ra-v on mouse lymphoid leukemia L1210, mm2 breast cancer; Ra- Ⅶ has obvious activity against mouse L1210, B16 melanoma, colon adenocarcinoma colon38, Lewis lung cancer, Ehrlich cancer, etc. In vivo tumor test in mice showed that rc-18, a monomer with anticancer activity extracted from the root of Herba eupatorii, could significantly prolong the life span of mice with P388, L1210 and B16, but had no obvious inhibitory effect on Lewis lung cancer and sarcoma S180. . In addition, 2-methoxycarbyl-3-isopentenyl-2,3-epoxy-1,4. naphthoquinone and menadione derivatives have anticancer activity against hamster lung (V79) cells, lymphoid leukemia (P388) and human nasopharyngeal carcinoma (KB) cells in vitro. It is effective for mouse ascites S180 in the overall test.

7. effect on urinary tract stones: 20% rubidium l preparation fed to mice can prevent the formation of experimental kidney and bladder stones, especially the formation of calcium carbonate stones. The results showed that the administration of Herba Siegesbeckiae extract to rats could significantly improve the stability of urine, reduce the risk of urinary stone formation, and have a certain effect of reducing urinary calcium.

8. therapeutic effect on experimental myocardial infarction: the artificial myocardial infarction model was created by continuous ligation of the large anterior descending branch of the left coronary artery. Alizarin Ⅰ and alizarin Ⅱ isolated by different methods were intravenously injected at a dose of 200mg/kg, once every 4 hours, for a total of 3 times, all of which could reduce the elevation of S-T segment and reduce the scope of myocardial infarction. Among them, Qian Ⅱ has the most obvious effect. Intraperitoneal injection of the water-soluble part of Rubia extract into mice can significantly increase the content of ATP in cardiac muscle and brain tissue, and has a depolymerizing effect on ADP induced platelet aggregation in rats. It has a protective effect on acute myocardial ischemia with large anesthesia, reduces the scope and extent of myocardial injury, and increases coronary flow.

9. effect on smooth muscle: qiancaogen decoction can resist isolated rabbit intestinal spasm caused by acetylcholine and has spasmolytic effect. .

10. other effects: Rubia root warm extract has the effect of expanding frog foot blood vessels. .