1、 Pharmacopoeia standard of Radix Isatidis
This product is the dried root of Isatis indigotica fort., a cruciferous plant. Excavate in autumn, remove sediment and dry in the sun.
[properties] this product is cylindrical, slightly twisted, 10 ~ 20cm long and 0.5 ~ LCM in diameter. The surface is light grayish yellow or light brownish yellow, with longitudinal wrinkles, transverse lenticel like protrusions and root marks. The root head is slightly expanded, and dark green or dark brown whorled petiole residues and dense warty protrusions can be seen. The body is solid and slightly soft, with yellowish white skin and yellow wood on the cross section. It tastes slightly sweet and bitter.
. The inner layer of the plug is narrow. Phloem is broad with obvious rays. The cambium forms a ring. Xylem vessels are yellow, quasi circular, with a diameter of about 80 μ m; There are wood fiber bundles. Parenchyma cells contain starch granules.
(2) Take 0.5g of this product powder, add 20ml of diluted ethanol, sonicate for 20 minutes, filter, evaporate the filtrate, add 1ml of diluted ethanol to dissolve the residue, and use it as the test solution. Another 0.5g of Radix Isatidis reference medicinal material was taken, and the reference medicinal material solution was prepared by the same method. Then take arginine reference substance and add diluted ethanol to make a solution containing 0.5mg per 1ml as the reference substance solution. . In the chromatogram of the test sample, spots with the same color appear at the corresponding positions of the chromatogram of the control medicinal material and the chromatogram of the control sample.
(3) Take 1g of this product powder, add 20ml of 80% methanol, sonicate for 30 minutes, filter, evaporate the filtrate, and add 1ml of methanol to dissolve the residue as the test solution. Another 1g of Radix Isatidis reference medicinal material was taken, and the reference medicinal material solution was prepared by the same method. . Test according to thin-layer chromatography (general rule 0502), suck 5-10 μ l of each of the above three solutions, dot them on the same silica gel gf254 thin-layer plate, develop them with petroleum ether (60-90 ℃) - ethyl acetate (1:1), take them out, dry them, and view them under UV light (254nm). In the chromatogram of the test sample, spots with the same color appear at the corresponding positions of the chromatogram of the control medicinal material and the chromatogram of the control sample.
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The total ash content shall not exceed 9.0% (general rule 2302).
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[extract] according to the hot leaching method under the alcohol soluble extract determination method (general rule 2201), 45% ethanol shall be used as solvent, which shall not be less than 25.0%.
[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-0.02% phosphoric acid solution (7:93); The detection wavelength is 245nm. The number of theoretical plates should not be less than 5000 according to (R, s) - guyichun peak.
Preparation of reference solution take an appropriate amount of (R, s) - gaoyichun reference, weigh accurately, add methanol to make a solution containing 40 μ g per 1ml.
Preparation of the test solution take about 1g of the powder (passing the No. 4 screen), accurately weigh it, place it in a round bottomed bottle, precisely add 50ml of water, weigh it, decoct it for 2 hours, cool it, weigh it again, make up the lost weight with water, shake it well, filter it, and take the filtrate.
The determination method is to precisely suck 10 ～ 20 μ l of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine.
The content of (R, s) - guayichun (c5h7nos) shall not be less than 0.020% according to the dry product.
Decoction pieces
[processing] remove impurities, wash, moisten, cut into thick pieces, and dry.
This product is a thick round piece. The outer skin is light grayish yellow to light brownish yellow, with longitudinal wrinkles. . It tastes slightly sweet and bitter.
[inspection] the moisture content is the same as that of medicinal materials, and shall not exceed 13.0%.
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[content determination] the content of (R, s) - gaoyichun (c5h7nos) in the same medicinal material shall not be less than 0.030%.
[identification] (except the cross-sectional surface) [inspection] (acid insoluble ash) [extract] is the same as that of medicinal materials.
[nature, taste and meridian tropism] bitter, cold. Homing and stomach meridian.
[functions and indications] it can clear away heat, detoxify, cool blood and facilitate swallowing. .
[usage and dosage] 9 ～ 15g.
[storage] put it in a dry place to prevent mold and moth.

2、 Chemical constituents of Radix Isatidis

Containing indigotin (Indigo),Indirubin（indirubin），β - sitosterol(β -sitosterol), γ - sitosterol, and a variety of amino acids:L-arginine（L-arginine）， Glutamic acid,l-tyrosine （L-tyrosine），L-Proline （L-proline），L-valine （L-valine），γ - aminobutyric acid（γ-aminobu-tyric acid）。 Also containsMyroside（sinigrin）， ,Report to Yichun（epigoitrin）， Adenosine,palmitic acid（palmitic acid）， Sucrose and proteoglycans containing 12% amino acids.

3、 Pharmacological effects of Radix Isatidis

1. antibacterial and antiviral effects: Isatis indigotica root has effects on a variety of bacteria. ; Acetone extract also has a similar effect and is effective against hemolytic streptococcus (both using agar small hole plate method). The antibacterial effect on type A meningococcus is similar to that of garlic and honeysuckle. Isatidis root and Folium Isatidis have antibacterial and antiviral active ingredients. Some people believe that they are indigo glycosides. The experiment proved that indioside had no antibacterial effect in vivo and in vitro; The antiviral effect in vitro was not prominent; It is excreted rapidly in the body, so it is considered that it has no obvious physiological and antibacterial activity. Its active ingredients remain to be studied.

2. anti Leptospira effect: Banlangen or Folium Isatidis above 1:100 can kill Leptospira in the test camp.

3. detoxification: it is reported that taking Radix Isatidis and Rhizoma Coptidis powder together with Veratrum (2.0g / kg each) for dogs can detoxify Veratrum and reduce mortality; If resveratrol is used after poisoning, it is invalid; The effect of using Radix Isatidis powder or Rhizoma Coptidis powder alone is also not good.

4. improve immune function: intraperitoneal injection of isatidis polysaccharide 50mg / kg in mice can significantly promote the immune function of mice, which is shown as: it can significantly increase the spleen weight of normal mice. The total number of white blood cells and lymphocytes had a significant effect on the decrease of spleen index, total number of white blood cells and lymphocyte number in immunosuppressed mice induced by hydrocortisone; Significantly enhanced the delayed anaphylaxis in normal and cyclophosphamide induced immunosuppressed mice induced by dinitrochlorobenzene; It enhanced the positive percentage of ANAE in peripheral blood lymphocytes of normal mice and significantly antagonized the immunosuppressive effect caused by hydrocortisone; However, Radix Isatidis polysaccharide did not significantly enhance the splenocyte lymphocytic transformation induced by concanavalin A in vitro. In addition, Radix Isatidis polysaccharide can significantly enhance the function of antibody forming cells and enhance the clearance rate of carbon particles in mice after intravenous injection.

5. antitumor effect: indirubin 200mg/kg was injected into rat W256 solid tumor and mouse Lewis lung cancer for 6-7 days. The results showed that the inhibition rates of rat W256 solid tumor were 47-50% and 50-58%, respectively. When subcutaneous injection was reduced to less than 100mg / kg per day, the inhibition effect was not obvious. Indirubin 500mg/kg by gavage has an inhibitory rate of 23-33% on rat W256 solid tumor and about 30% on mouse sarcoma 180. Indirubin can prolong the survival time of l7212 mice with lymphoid leukemia by about 20%. In addition, the mouse spermatogonial assay showed that indirubin could not specifically inhibit B-type spermatogonia and spermatids.

6. effect on leukemia: pharmacological studies preliminarily showed that indirubin has the effect of destroying leukemia cells. According to the ultrastructural morphology, most of the degenerated and necrotic cells showed swelling and lytic necrosis under the action of indirubin. In the experiment, indirubin was found to enhance the phagocytic capacity of animal mononuclear macrophage system. The mononuclear macrophagy system plays a certain role in the body's immune response, so the anticancer effect of indirubin may be related to improving the body's immune ability.