根据《中国药典·2020年版·一部》记载：本天然药材以牛科动物牛Bos taurus domesticus Gmelin的新鲜胆汁作母液，加入去氧胆酸、胆酸、复合胆红素钙等制成。
According to the Chinese Pharmacopoeia: 2020 edition: Volume I, this natural medicinal material is processed from fresh bile of the bovine animal Bos taurus domesticus Gmelin, with deoxycholic acid, cholic acid, complex bilirubin calcium, etc.
NMM ID | nmm-002d |
系统名 | In Vitro Cultivated Calculus Bovis |
系统中文名 | 体外培育牛黄（tǐ wài péi yù niú huáng） |
通用名 | Ti-wai-pei-yu-niu-huang |
通用中文名 | 体外培育牛黄（tǐ wài péi yù niú huáng） |
类型 | artificial |
物种基源 | |
药用部位 | |
特殊描述 | |
炮制方法 | |
系统名命名解释 | 根据《中国药典·2020年版·一部》记载：本天然药材以牛科动物牛Bos taurus domesticus Gmelin的新鲜胆汁作母液，加入去氧胆酸、胆酸、复合胆红素钙等制成。 According to the Chinese Pharmacopoeia: 2020 edition: Volume I, this natural medicinal material is processed from fresh bile of the bovine animal Bos taurus domesticus Gmelin, with deoxycholic acid, cholic acid, complex bilirubin calcium, etc. |
通用名命名解释 | NMMGN衍生自《中国药典·2020年版·一部》相关中药材中文名。 |
上级药材 | |
下级药材 | |
创建人 | |
审核专家 |
体外培育牛黄
中文文本参考：《中国药典（2020年版）》
English text reference: Chinese Pharmacopoeia (2020 Edition)
概述 Overview
本品以牛科动物牛Bos taurus domesticus Gmelin的新鲜胆汁作母液，加入去氧胆酸、胆酸、复合胆红素钙等制成。
Artificially Cultivated Calculus Bovis is the concentrated fresh bile of Bos taurus domesticus Gmelin (Fam. Bovidae), mixed with deoxycholic acid, cholic acid, and calcium bilirubinate.
性状 Description
本品呈球形或类球形，直径0.5～3 cm。表面光滑，呈黄红色至棕黄色。体轻，质松脆，断面有同心层纹。气香，味苦而后甘，有清凉感，嚼之易碎，不粘牙。
Calculus Bovis Sativus is spherical or nearly spherical, measuring 0.5-3 cm in diameter. The surface is smooth and yellowish-red to brownish-yellow. It is light in weight, brittle in texture, and shows concentric layers on the fractured surface. It has a characteristic aroma, a bitter taste followed by sweetness, and imparts a cooling sensation. It is easily broken when chewed and does not adhere to the teeth.
鉴别 Identification
（1）取本品粉末少量，用清水调和，涂于指甲上，能将指甲染成黄色。
(1) Take a small amount of the powder, mix it with water, and apply it to the fingernails. It should turn the fingernails yellow.
（2）取本品粉末少许，用水合氯醛试液装片，不加热，置显微镜下观察：不规则团块由多数黄棕色或棕红色小颗粒集成，稍放置，色素迅速溶解，并显鲜明金黄色，久置后变绿色。
(2) Take a small amount of the powder, prepare a slide with chloral hydrate solution, and observe it under a microscope without heating: irregular aggregates composed of numerous yellow-brown or brownish-red small particles can be seen. After a short period of time, the pigment dissolves rapidly and appears as a bright golden yellow color, which turns green after a long period of time.
（3）取本品粉末少量，加三氯甲烷1 ml，摇匀，再加硫酸与浓过氧化氢溶液（30%）各2滴，振摇，溶液即显绿色。
(3) Take a small amount of the powder, add 1 ml of chloroform, shake well, then add 2 drops of sulfuric acid and concentrated hydrogen peroxide solution (30%), shake well, and the solution should turn green.
（4）取本品粉末0.1 g，加盐酸1 ml和三氯甲烷10 ml，充分振摇，混匀，三氯甲烷液呈黄褐色，分取三氯甲烷液，加氢氧化钡试液5 ml，振摇，即生成黄褐色沉淀。分离除去水层和沉淀，取三氯甲烷液约1 ml，加醋酐1 ml与硫酸2滴，摇匀，放置，溶液呈绿色。
(4) Take 0.1 g of the powder, add 1 ml of hydrochloric acid and 10 ml of chloroform, shake vigorously to mix well, the chloroform solution should be yellowish-brown. Take a portion of the chloroform solution, add 5 ml of barium hydroxide test solution, shake well, and a yellowish-brown precipitate should form. Separate the water layer and the precipitate, take about 1 ml of the chloroform solution, add 1 ml of acetic anhydride and 2 drops of sulfuric acid, shake well, let it stand, and the solution should turn green.
（5）取本品粉末10 mg，加三氯甲烷20 ml，超声处理30分钟，滤过，滤液蒸干，残渣加乙醇1 ml使溶解，作为供试品溶液。另取胆酸对照品、去氧胆酸对照品，加乙醇制成每1 ml各含2 mg的混合溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各2 μl，分别点于同一硅胶G 薄层板上，以异辛烷-乙酸乙酯-冰醋酸（15：7：5）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，在105°C加热至斑点显色清晰，置紫外光灯（365 nm）下检视。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的荧光斑点。
(5) Take 10 mg of the powder, add 20 ml of chloroform, and sonicate for 30 minutes. Filter the solution, evaporate the filtrate to dryness, and dissolve the residue in 1 ml of ethanol as the test solution. Take cholic acid reference substance and deoxycholic acid reference substance, add ethanol to prepare a mixed solution containing 2 mg of each substance per 1 ml as the reference solution. Perform thin-layer chromatography<0502> using silica gel G as the coating substance and a mixture of isooctane, ethyl acetate, and glacial acetic acid (15:7:5) as the mobile phase. Apply separately to the plate 2 μl of each of the above two solutions. After developing and removal of the plate, dry in air. Spray with a 10% solution of sulfuric acid in ethanol, heat at 105°C to the spots clear. Examine under ultraviolet light at 365 nm. The fluorescent spot in the chromatogram obtained with the test solution corresponds in position and colour to the spot in the chromatogram obtained with the reference solution.
检查 Examination
水分 Water
不得过9.0%（通则0832第二法）。
Not more than 9.0 per cent <0832,method 2>.
游离胆红素 Free bilirubin
取本品细粉10 mg，精密称定，置5 ml量瓶中，加三氯甲烷4 ml，微温，放冷，加三氯甲烷至刻度，摇匀，滤过，取续滤液，照紫外-可见分光光度法（通则0401），在453 nm 波长处测定吸光度。吸光度不得过0.70。
Take 10 mg of the powder, accurately weigh, place it in a 5 ml volumetric flask, add 4 ml of chloroform, warm slightly, cool, add chloroform to the mark, shake well, filter, take the filtrate, and determine the absorbance at a wavelength of 453 nm by ultraviolet-visible spectrophotometry. The absorbance should not exceed 0.70.
含量测定 Content determination
胆酸 Cholic acid
取本品细粉0.2 g，精密称定，置具塞锥形瓶中，精密加入甲醇50 ml，密塞，称定重量，超声处理30分钟，放冷，再称定重量，用甲醇补足减失的重量，摇匀，滤过。精密量取续滤液25 ml，蒸干，残渣加20%氢氧化钠溶液10 ml，加热回流2小时，冷却，加稀盐酸19 ml调节pH值至酸性，用乙酸乙酯提取4次（25 ml、25 ml、20 ml、20 ml），乙酸乙酯液均用同一铺有少量无水硫酸钠的脱脂棉滤过，滤液合并，回收溶剂至干，残渣用甲醇溶解，转移至10 ml量瓶中，加甲醇至刻度，摇匀，作为供试品溶液。另取胆酸对照品适量，精密称定，加甲醇制成每1 ml含0. 48 mg的溶液，作为对照品溶液。 照薄层色谱法（通则0502）试验，精密吸取供试品溶液2 μl、对照品溶液1 μl与3 μl，分别交叉点于同一硅胶G薄层板上，以异辛烷-乙酸丁酯-冰醋酸-甲酸（8：4：2：1）为展开剂，展距14～17 cm，取出，晾干，喷以30%硫酸乙醇溶液，在105°C加热至斑点显色清晰，取出，在薄层板上覆盖同样大小的玻璃板，周围用胶布固定，照薄层色谱法（通则0502薄层色谱扫描法）进行扫描，波长：λs = 380 nm，λR = 650 nm，测量供试品吸光度积分值与对照品吸光度积分值，计算，即得。
Take 0.2 g of the powder, accurately weigh, place it in a stoppered conical flask, accurately add 50 ml of methanol, seal tightly, weigh, treat with ultrasound for 30 minutes, cool, weigh again, make up for the weight loss with methanol, shake well, filter. Precisely take 25 ml of the filtrate, evaporate to dryness, add 10 ml of 20% sodium hydroxide solution to the residue, heat reflux for 2 hours, cool, add 19ml of dilute hydrochloric acid to adjust the pH to acidic, extract with ethyl acetate 4 times (25 ml, 25 ml, 20 ml, 20 ml), filter the ethyl acetate solution with the same amount of anhydrous sodium sulfate-degreased cotton, combine the filtrate, recover the solvent to dryness, dissolve the residue with methanol, transfer to a 10 ml volumetric flask, add methanol to the mark, shake well, and use as the test solution. Take an appropriate amount of cholic acid reference substance, accurately weigh, add methanol to make a solution containing 0.48 mg per 1 ml, and use as the reference solution. Perform a thin-layer chromatography test according to the method specified in <0502>, precisely take 2 μl, 1 μl, and 3 μl of the test solution and the reference solution, respectively, and spot them on the same silica gel G thin-layer plate. Use n-hexane-ethyl acetate-glacial acetic acid-formic acid (8:4:2:1) as the developing agent, with a development distance of 14-17 cm. Take out the plate, dry it, spray it with a 30% sulfuric acid-ethanol solution, heat it at 105°C until the spots are clearly visible, take it out, cover the plate with a glass plate of the same size, fix it with adhesive tape, and scan it by thin-layer chromatography (TLC) scanning method specified in <0502>. Wavelength: λs = 380 nm, λR = 650 nm. Measure the integrated absorbance of the test solution and the reference solution, and calculate accordingly.
本品按干燥品计算，含胆酸（C24 H40O5）不得少于6.0%。
This product contains not less than 6.0% of bile acid (C24 H40O5) calculated as dried product.
胆红素 Bilirubin
对照品溶液的制备 Preparation of control solution
取胆红素对照品10 mg，精密称定，置100 ml棕色量瓶中，用三氯甲烷溶解并稀释至刻度，摇匀，精密量取5 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀，即得（每1 ml中含胆红素10 μg）。
Take 10 mg of bilirubin control, weigh it precisely, put it into 100 ml brown measuring flask, dissolve and dilute it to the scale with trichloromethane, shake well, measure 5 ml precisely, put it into 50 ml brown measuring flask, add ethanol to the scale, shake well, that is to say, it is obtained (each 1 ml contains 10 μg of bilirubin).
标准曲线的制备 Preparation of standard curves
精密量取对照品溶液1 ml、2 ml、3 ml、4 ml、5 ml，分别置具塞试管中，加乙醇至9 ml，各精密加入重氮化溶液（甲液：取对氨基苯磺酸0. 1 g，加盐酸，1.5 ml与水适量使成100 ml。乙液：取亚硝酸钠0.5 g，用水溶解并稀释至100 ml，置冰箱内保存。临用时取甲液10 ml与乙液0.3 ml，混匀）1 ml,摇匀，在15～20°C的暗处放置1小时，以相应的试剂为空白，照紫外-可见分光光度法（通则0401），在533 nm波长处测定吸光度，以吸光度为纵坐标、浓度为横坐标，绘制标准曲线。
Precisely measure 1 ml, 2 ml, 3 ml, 4 ml, 5 ml of control solution, respectively, placed in a stoppered test tube, add ethanol to 9 ml, add diazotisation solution (solution A: take 0.1 g of p-aminobenzenesulphonic acid, add hydrochloric acid, 1.5 ml and water to make 100 ml; solution B: take 0.5 g of sodium nitrite, dissolve with water and dilute to 100 ml, keep in the refrigerator. At the time of use, take 10 ml of A solution and 0.3 ml of B solution, mix well) 1 ml, shake well, place in the dark at 15-20°C for 1 hour, use the corresponding reagent as the blank, according to the UV-visible spectrophotometric method (General rule 0401), determine the absorbance at the wavelength of 533 nm, take the absorbance as the vertical coordinate, the concentration as the horizontal coordinate, draw the standard curve.
测定法 Method of determination
取本品细粉约10 mg，精密称定，置锥形瓶中，加三氯甲烷-乙醇（7：3）的混合溶液60 ml、盐酸1滴，摇匀，置水浴上加热回流30分钟，放冷，转移至100 ml棕色量瓶中，容器用少量上述混合溶液洗涤，洗液并入同一量瓶中，加上述混合溶液至刻度，摇匀。精密量取上清液10 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀。精密量取3 ml，置具塞试管中，照标准曲线的制备项下的方法，自“加乙醇至9 ml”起，依法测定吸光度，从标准曲线上读岀供试品溶液中含胆红素的重量（mg），计算，即得。
Take about 10 mg of this product fine powder, weighed precisely, placed in a conical flask, add 60 ml of trichloromethane-ethanol (7:3) mixed solution, hydrochloric acid 1 drop, shaking well, placed in a water bath heated reflux for 30 minutes, cooled, and transferred to a 100 ml brown measuring flask, the container was washed with a small amount of the above mixture of solutions, washings were incorporated into the same measuring flask, add the above mixture of solutions to the graduated scale, shaking well. Precisely measure 10 ml of supernatant, put in 50 ml brown measuring flask, add ethanol to the scale, shake well. Precisely measure 3 ml, put in a stoppered test tube, according to the method under the preparation of the standard curve, since "add ethanol to 9 ml", according to the law to determine the absorbance, from the standard curve read out the weight of bilirubin in the test solution (mg), calculated, that is, obtained.
本品按干燥品计算，含胆红素（C33 H36 N4O6）不得少于35.0%。
This product contains not less than 35.0% of bilirubin (C33 H36 N4O6) calculated as dried product.
性 Property
凉。
Cold.
毒性 Toxicity
无。
None.
味 Flavor
甘。
Sweet.
归经 Meridian tropism
心、肝经。
Heart and liver meridians.
功能 Actions
清心，豁痰，开窍，凉肝，息风，解毒。
To clear the heart, resolve phlegm, open the orifices, cool the liver, relieve wind, and detoxify.
主治 Indications
用于热病神昏，中风痰迷，惊痫抽搐，癫痫发狂，咽喉肿痛，口舌生疮，痈肿疔疮。
Used for coma due to febrile diseases, stroke with phlegm obstruction, convulsions of epilepsy, mania of epilepsy, sore throat, oral ulcers, carbuncles and furuncles.
用量 Dosage
0.15～0.35 g。
0.15-0.35 g.
用法 Administration
多入丸散用。外用适量，研末敷患处。
taken in pill or powder form. For external use, apply an appropriate amount of powdered medicine to the affected area.
注意 Precautions
孕妇慎用；偶有轻度消化道不适。
Caution for pregnant women; occasional mild gastrointestinal discomfort.
贮藏 Storage
密闭，遮光，防潮，防压，室温保存。
Store in a sealed, light-proof, moisture-proof, and pressure-resistant container at room temperature.
附：
去氧胆酸质量标准 Deoxycholic acid quality standard
去氧胆酸 Deoxycholic acid
本品由牛胆汁经提取、加工制成。
This product is extracted and processed from bovine bile.
性状 Description
本品为白色的结晶性粉末。气微，味微苦。
This product is a white crystalline powder. It has a slight odor and a slightly bitter taste.
本品易溶于冰醋酸和乙醇，不溶于水。
This product is soluble in glacial acetic acid and ethanol, insoluble in water.
鉴别 Identification
取本品10 mg，加乙醇5 ml使溶解，作为供试品溶液。另取去氧胆酸对照品，加乙醇制成每1 ml含2 mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各2 μl，分别点于同一硅胶G薄层板上，以异辛烷-乙酸乙酯-冰醋酸（15：7：5）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，在105°C加热至斑点显色清晰，置紫外光灯（365 nm）下检视。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的荧光斑点。
Take 10 mg of the product, dissolve it in 5 ml of ethanol, and use it as the test solution. Take an appropriate amount of deoxycholic acid reference substance, dissolve it in ethanol to make a solution containing 2 mg per 1 ml, and use it as the reference solution. Perform a thin-layer chromatography test according to the method specified in <0502>, take 2 μl of each of the above-mentioned solutions, spot them on the same silica gel G thin-layer plate, use n-hexane-ethyl acetate-glacial acetic acid (15:7:5) as the developing agent, develop, take out, dry, spray with a 10% sulfuric acid-ethanol solution, heat at 105°C until the spots are clearly visible, and examine under ultraviolet light (365nm). In the chromatogram of the test solution, there should be fluorescent spots of the same color at the corresponding positions as in the chromatogram of the reference solution.
检查 Examination
干燥失重 Loss on drying
取本品，在105°C干燥4小时，减失重量不得过0.5%（通则0831）。
Take the product, dry it at 105°C for 4 hours, and the loss on drying should not exceed 0.5% <0831>.
炽灼残渣 Residue on ignition
不得过0.2%（通则0841）。
The residue on ignition should not exceed 0.2% <0841>.
含量测定 Content determination
取本品约0. 5 g，精密称定，加中性乙醇（对酚酞指示液显中性）60 ml，置水浴中加热使溶解，冷却，加酚酞指示液数滴及新沸过的冷水20 ml，用氢氧化钠滴定液（0.1 mol/L）滴定，近终点时加新沸过的冷水100 ml，继续滴定至终点。每1ml氢氧化钠滴定液（0.1 mol/L）相当于39. 26 mg 的去氧胆酸（C24 H40O4）。
Take about 0.5g of the product, accurately weigh, place it in a 250 ml conical flask, add 20 ml of water and 40 ml of neutral ethanol (the phenolphthalein indicator solution shows neutrality), heat on a water bath to dissolve, cool, add a few drops of phenolphthalein indicator solution and 20 ml of freshly boiled and cooled water, titrate with sodium hydroxide titration solution (0.1 mol/L) until near the end point, add 100 ml of freshly boiled and cooled water, and continue titration to the end point. Each 1ml of sodium hydroxide titration solution (0.1 mol/L) is equivalent to 39.26 mg of deoxycholic acid (C24H40O4).
本品按干燥品计算，含去氧胆酸（C24 H40O4）不得少于95.0%。
Calculated on the dried basis, the content of deoxycholic acid (C24H40O4) should not be less than 95.0%.
用途 Usage
体外培育牛黄的原料。
Raw material for in vitro cultivation of oxalis.
贮藏 Storage
密封。
Sealing.
胆酸质量标准 Cholic acid quality standard
胆 酸 Cholic acid
本品由牛、羊胆汁或胆膏经提取、加工制成。
This product is extracted and processed from bovine or sheep bile or bile paste.
性状 Description
本品为白色的结晶性粉末。气微，味苦。
This product is a white crystalline powder. It has a slight odor and a bitter taste.
鉴别 Identification
取本品10 mg，加乙醇5 ml使溶解，作为供试品溶液。另取胆酸对照品，加乙醇制成每1 ml含2 mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各2 μl，分别点于同一硅胶G薄层板上，以异辛烷-乙酸乙酯-冰醋酸（15：7：5）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，在105°C加热至斑点显色清晰，置紫外光灯（365nm）下检视。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的荧光斑点。
Take 10 mg of the product, dissolve it in 5 ml of ethanol, and use it as the test solution. Take an appropriate amount of cholic acid reference substance, dissolve it in ethanol to make a solution containing 2 mg per 1 ml, and use it as the reference solution. Perform a thin-layer chromatography test according to the method specified in <0502>, take 2 μl of each of the above-mentioned solutions, spot them on the same silica gel G thin-layer plate, use n-hexane-ethyl acetate-glacial acetic acid (15:7:5) as the developing agent, develop, take out, and dry. Spray with 10% ethanol sulphate solution, heat at 105°C until the spots show clear colour, and examine under UV light (365 nm). In the chromatogram of the test product, in the corresponding position with the chromatogram of the control product, the fluorescent spots of the same colour are shown.
检查 Examination
干燥失重 Loss on drying
取本品，在105°C干燥4小时，减失重量不得过0.5%（通则0831）。
Take the product, dry it at 105°C for 4 hours, and the loss on drying should not exceed 0.5% <0831>.
炽灼残渣 Residue on ignition
不得过0.2%（通则0841）。
The residue on ignition should not exceed 0.2% <0841>.
含量测定 Content determination
取本品约0.4 g，精密称定，置250 ml锥形瓶中，加水20 ml和乙醇40 ml，用表面皿覆盖，置水浴中缓缓加热使完全溶解，冷却，加酚酞指示液5滴，用氢氧化钠滴定液（0. 1 mol/L）滴定。每1 ml氢氧化钠滴定液（0.1 mol/L）相当于40.86 mg的胆酸（C24H40O24）。
Take about 0.4 g of the product, accurately weigh, place it in a 250 ml conical flask, add 20 ml of water and 40 ml of ethanol, cover with a watch glass, heat slowly on a water bath to completely dissolve, cool, add 5 drops of phenolphthalein indicator solution, and titrate with sodium hydroxide titration solution (0.1 mol/L). Each 1 ml of sodium hydroxide titration solution (0.1 mol/L) is equivalent to 40.86 mg of cholic acid (C24H40O24).
本品按干燥品计算，含胆酸（C24H40O24）不得少于95.0%。
Calculated on the dried basis, the content of cholic acid (C24H40O24) should not be less than 95.0%.
用途 Usage
体外培育牛黄的原料。
Raw material for in vitro cultivation of oxalis.
贮藏 Storage
密封。
Sealing.
复合胆红素钙质量标准 Compound Bilirubin Calcium quality standard
复合胆红素钙 Compound Bilirubin Calcium
本品由牛胆汁、胆红素和饱和氢氧化钙溶液经加工制成。
This product is processed from bovine bile, bilirubin, and saturated calcium hydroxide solution.
性状 Description
本品为棕红色或棕黄色的粉末。味微腥。
This product is a brownish-red or brownish-yellow powder. It has a slightly fishy taste.
鉴别 Identification
取本品粉末5 mg，加三氯甲烷15 ml，加盐酸溶液（5→10）0. 1 ml，超声处理30分钟，加三氯甲烷35 ml，摇匀，滤过，取续滤液作为供试品溶液。另取胆红素对照品，加三氯甲烷制成每1 ml含0.1 mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各10 μl，分别点于同一硅胶G薄层板上，以甲苯-乙酸乙酯-冰醋酸（10：1：0.5）为展开剂，展开，取出，晾干。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的斑点。
Take 5 mg of this product powder, add 15 ml of trichloromethane, add hydrochloric acid solution (5→10) 0.1 ml, ultrasonic treatment for 30 minutes, add 35 ml of trichloromethane, shaking well, filtration, take the filtrate as the test solution. Take bilirubin control product, add trichloromethane to make a solution containing 0.1 mg per 1 ml, as the control solution. According to the thin-layer chromatography (General rule 0502) test, absorb the above two solutions of 10 μl, were spotted on the same silica gel G thin-layer plate, toluene - ethyl acetate - glacial acetic acid (10: 1: 0.5) as an unfolding agent, unfolding, remove, and dry. In the chromatogram of the test article, spots of the same colour are shown in the corresponding positions with the chromatogram of the control article.
检查 Examination
水分 Water
不得过5.0%（通则0832第二法）。
Not more than 5.0 per cent (General Law 0832, second law).
含量测定 Content determination
对照品溶液的制备 Preparation of control solution
取胆红素对照品 10 mg，精密称定，置100 ml棕色量瓶中，用三氯甲烷溶解并稀释至刻度，摇匀。精密量取5 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀，即得（每1 ml中含胆红素10 μg）。
Take 10 mg of bilirubin control product, weigh it precisely, put it in 100 ml brown measuring flask, dissolve and dilute it with trichloromethane to the scale, shake well. Measure 5 ml, put in 50ml brown measuring flask, add ethanol to the scale, shake well, it is obtained (each 1 ml contains 10 μg of bilirubin).
标准曲线的制备 Preparation of standard curves
精密量取对照品溶液1 ml、2 ml、3 ml、4 ml、5 ml，分别置具塞试管中，加乙醇至9 ml，各精密加入重氮化溶液（甲液：取对氨基苯磺酸0.1 g，加盐酸1.5 ml与水适量使成100 ml。乙液：取亚硝酸钠0.5 g，用水溶解并稀释至100 ml，置冰箱内保存。临用时，取甲液10 ml与乙液0.3 ml，混匀）1 ml，摇匀，在15～20°C的暗处放置1小时，以相应的试，剂为空白，照紫外-可见分光光度法（通则0401），在533 nm波长处测定吸光度，以吸光度为纵坐标、浓度为横坐标，绘制标准曲线。
Precisely measure 1 ml, 2 ml, 3 ml, 4 ml, 5 ml of the control solution, respectively, placed in a stoppered test tube, add ethanol to 9 ml, add diazotisation solution (solution A: take 0.1 g of p-aminobenzenesulphonic acid, add 1.5 ml of hydrochloric acid and the appropriate amount of water to make 100 ml; solution B: take 0.5 g of sodium nitrite, dissolve with water and dilute to 100 ml, stored in the refrigerator. At the time of use, take 10 ml of solution A and 0.3 ml of solution B, mix well) 1 ml, shake well, and leave it in the dark at 15-20°C for 1 hour, take the corresponding test agent as blank, according to the UV-visible spectrophotometric method (General rule 0401), determine the absorbance at the wavelength of 533 nm, and plot a standard curve with the absorbance as the vertical coordinates and the concentration as the horizontal coordinates.
测定法 Method of determination
取本品细粉，约10 mg，精密称定，置锥形瓶中，加三氯甲烷-乙醇（7 ：3）的混合溶液60 ml、盐酸1滴，摇匀，置水浴上加热回流30分钟，放冷，转移至100 ml棕色量瓶中，容器用少量上述混合溶液洗涤，洗液并入同一量瓶中，加上述混合溶液至刻度，摇匀。精密量取上清液10 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀。精密量取3 ml，置具塞试管中，照标准曲线的制备项下的方法，自“加乙醇至9 ml”起，依法测定吸光度，从标准曲线上读出供试品溶液中含胆红素的重量（mg）,计算，即得。
Take the fine powder of this product, about 10 mg, weighed accurately, placed in a conical flask, add 60 ml of trichloromethane-ethanol (7:3) mixed solution, hydrochloric acid 1 drop, shake well, placed in a water bath heated reflux for 30 minutes, cooled, and transferred to a 100 ml brown measuring flask, the container was washed with a small amount of the above mixture of solutions, washings were incorporated into the same measuring flask, plus the above mixture of solutions to the graduated scale, shaking well. Precisely measure 10 ml of supernatant, put in 50 ml brown measuring flask, add ethanol to the scale, shake well. Precisely measure 3 ml, put in a stoppered test tube, according to the method under the preparation of the standard curve, since "add ethanol to 9 ml", according to the law to determine the absorbance, from the standard curve to read out the weight of bilirubin in the test solution (mg), calculation, that is, obtained.
本品按干燥品计算，含胆红素（C33H36 N4O6）不得少于43.0%。
This product contains not less than 43.0% of bilirubin (C33H36 N4O6) calculated as dry product.
用途 Usage
体外培育牛黄的原料。
Raw material for in vitro cultivation of oxalis.
贮藏 Storage
密封。
Sealing.
体外培育牛黄
中文文本参考：《中国药典（2015年版）》
English text reference: Chinese Pharmacopoeia (2020 Edition)
概述 Overview
本品以牛科动物牛Bos taurus domesticus Gmelin的新鲜胆汁作母液，加入去氧胆酸、胆酸、复合胆红素钙等制成。
Artificially Cultivated Calculus Bovis is the concentrated fresh bile of Bos taurus domesticus Gmelin (Fam. Bovidae), mixed with deoxycholic acid, cholic acid, and calcium bilirubinate.
性状 Description
本品呈球形或类球形，直径0.5～3 cm。表面光滑，呈黄红色至棕黄色。体轻，质松脆，断面有同心层纹。气香，味苦而后甘，有清凉感，嚼之易碎，不粘牙。
Calculus Bovis Sativus is spherical or nearly spherical, measuring 0.5-3 cm in diameter. The surface is smooth and yellowish-red to brownish-yellow. It is light in weight, brittle in texture, and shows concentric layers on the fractured surface. It has a characteristic aroma, a bitter taste followed by sweetness, and imparts a cooling sensation. It is easily broken when chewed and does not adhere to the teeth.
鉴别 Identification
（1）取本品粉末少量，用清水调和，涂于指甲上，能将指甲染成黄色。
(1) Take a small amount of the powder, mix it with water, and apply it to the fingernails. It should turn the fingernails yellow.
（2）取本品粉末少许，用水合氯醛试液装片，不加热，置显微镜下观察：不规则团块由多数黄棕色或棕红色小颗粒集成，稍放置，色素迅速溶解，并显鲜明金黄色，久置后变绿色。
(2) Take a small amount of the powder, prepare a slide with chloral hydrate solution, and observe it under a microscope without heating: irregular aggregates composed of numerous yellow-brown or brownish-red small particles can be seen. After a short period of time, the pigment dissolves rapidly and appears as a bright golden yellow color, which turns green after a long period of time.
（3）取本品粉末少量，加三氯甲烷1 ml，摇匀，再加硫酸与浓过氧化氢溶液（30%）各2滴，振摇，溶液即显绿色。
(3) Take a small amount of the powder, add 1 ml of chloroform, shake well, then add 2 drops of sulfuric acid and concentrated hydrogen peroxide solution (30%), shake well, and the solution should turn green.
（4）取本品粉末0.1 g，加盐酸1 ml和三氯甲烷10 ml，充分振摇，混匀，三氯甲烷液呈黄褐色，分取三氯甲烷液，加氢氧化钡试液5 ml，振摇，即生成黄褐色沉淀。分离除去水层和沉淀，取三氯甲烷液约1 ml，加醋酐1 ml与硫酸2滴，摇匀，放置，溶液呈绿色。
(4) Take 0.1 g of the powder, add 1 ml of hydrochloric acid and 10 ml of chloroform, shake vigorously to mix well, the chloroform solution should be yellowish-brown. Take a portion of the chloroform solution, add 5 ml of barium hydroxide test solution, shake well, and a yellowish-brown precipitate should form. Separate the water layer and the precipitate, take about 1 ml of the chloroform solution, add 1 ml of acetic anhydride and 2 drops of sulfuric acid, shake well, let it stand, and the solution should turn green.
（5）取本品粉末10 mg，加三氯甲烷20 ml，超声处理30分钟，滤过，滤液蒸干，残渣加乙醇1 ml使溶解，作为供试品溶液。另取胆酸对照品、去氧胆酸对照品，加乙醇制成每1 ml各含2 mg的混合溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各2 μl，分别点于同一硅胶G 薄层板上，以异辛烷-乙酸乙酯-冰醋酸（15：7：5）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，在105°C加热至斑点显色清晰，置紫外光灯（365 nm）下检视。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的荧光斑点。
(5) Take 10 mg of the powder, add 20 ml of chloroform, and sonicate for 30 minutes. Filter the solution, evaporate the filtrate to dryness, and dissolve the residue in 1 ml of ethanol as the test solution. Take cholic acid reference substance and deoxycholic acid reference substance, add ethanol to prepare a mixed solution containing 2 mg of each substance per 1 ml as the reference solution. Perform thin-layer chromatography<0502> using silica gel G as the coating substance and a mixture of isooctane, ethyl acetate, and glacial acetic acid (15:7:5) as the mobile phase. Apply separately to the plate 2 μl of each of the above two solutions. After developing and removal of the plate, dry in air. Spray with a 10% solution of sulfuric acid in ethanol, heat at 105°C to the spots clear. Examine under ultraviolet light at 365 nm. The fluorescent spot in the chromatogram obtained with the test solution corresponds in position and colour to the spot in the chromatogram obtained with the reference solution.
检查 Examination
水分 Water
不得过9.0%（通则0832第二法）。
Not more than 9.0 per cent <0832,method 2>.
游离胆红素 Free bilirubin
取本品细粉10 mg，精密称定，置5 ml量瓶中，加三氯甲烷4 ml，微温，放冷，加三氯甲烷至刻度，摇匀，滤过，取续滤液，照紫外-可见分光光度法（通则0401），在453 nm 波长处测定吸光度。吸光度不得过0.70。
Take 10 mg of the powder, accurately weigh, place it in a 5 ml volumetric flask, add 4 ml of chloroform, warm slightly, cool, add chloroform to the mark, shake well, filter, take the filtrate, and determine the absorbance at a wavelength of 453 nm by ultraviolet-visible spectrophotometry. The absorbance should not exceed 0.70.
含量测定 Content determination
胆酸 Cholic acid
取本品细粉0.2 g，精密称定，置具塞锥形瓶中，精密加入甲醇50 ml，密塞，称定重量，超声处理30分钟，放冷，再称定重量，用甲醇补足减失的重量，摇匀，滤过。精密量取续滤液25 ml，蒸干，残渣加20%氢氧化钠溶液10 ml，加热回流2小时，冷却，加稀盐酸19 ml调节pH值至酸性，用乙酸乙酯提取4次（25 ml、25 ml、20 ml、20 ml），乙酸乙酯液均用同一铺有少量无水硫酸钠的脱脂棉滤过，滤液合并，回收溶剂至干，残渣用甲醇溶解，转移至10 ml量瓶中，加甲醇至刻度，摇匀，作为供试品溶液。另取胆酸对照品适量，精密称定，加甲醇制成每1 ml含0. 48 mg的溶液，作为对照品溶液。 照薄层色谱法（通则0502）试验，精密吸取供试品溶液2 μl、对照品溶液1 μl与3 μl，分别交叉点于同一硅胶G薄层板上，以异辛烷-乙酸丁酯-冰醋酸-甲酸（8：4：2：1）为展开剂，展距14～17 cm，取出，晾干，喷以30%硫酸乙醇溶液，在105°C加热至斑点显色清晰，取出，在薄层板上覆盖同样大小的玻璃板，周围用胶布固定，照薄层色谱法（通则0502薄层色谱扫描法）进行扫描，波长：λs = 380 nm，λR = 650 nm，测量供试品吸光度积分值与对照品吸光度积分值，计算，即得。
Take 0.2 g of the powder, accurately weigh, place it in a stoppered conical flask, accurately add 50 ml of methanol, seal tightly, weigh, treat with ultrasound for 30 minutes, cool, weigh again, make up for the weight loss with methanol, shake well, filter. Precisely take 25 ml of the filtrate, evaporate to dryness, add 10 ml of 20% sodium hydroxide solution to the residue, heat reflux for 2 hours, cool, add 19ml of dilute hydrochloric acid to adjust the pH to acidic, extract with ethyl acetate 4 times (25 ml, 25 ml, 20 ml, 20 ml), filter the ethyl acetate solution with the same amount of anhydrous sodium sulfate-degreased cotton, combine the filtrate, recover the solvent to dryness, dissolve the residue with methanol, transfer to a 10 ml volumetric flask, add methanol to the mark, shake well, and use as the test solution. Take an appropriate amount of cholic acid reference substance, accurately weigh, add methanol to make a solution containing 0.48 mg per 1 ml, and use as the reference solution. Perform a thin-layer chromatography test according to the method specified in <0502>, precisely take 2 μl, 1 μl, and 3 μl of the test solution and the reference solution, respectively, and spot them on the same silica gel G thin-layer plate. Use n-hexane-ethyl acetate-glacial acetic acid-formic acid (8:4:2:1) as the developing agent, with a development distance of 14-17 cm. Take out the plate, dry it, spray it with a 30% sulfuric acid-ethanol solution, heat it at 105°C until the spots are clearly visible, take it out, cover the plate with a glass plate of the same size, fix it with adhesive tape, and scan it by thin-layer chromatography (TLC) scanning method specified in <0502>. Wavelength: λs = 380 nm, λR = 650 nm. Measure the integrated absorbance of the test solution and the reference solution, and calculate accordingly.
本品按干燥品计算，含胆酸（C24 H40O5）不得少于6.0%。
This product contains not less than 6.0% of bile acid (C24 H40O5) calculated as dried product.
胆红素 Bilirubin
对照品溶液的制备 Preparation of control solution
取胆红素对照品10 mg，精密称定，置100 ml棕色量瓶中，用三氯甲烷溶解并稀释至刻度，摇匀，精密量取5 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀，即得（每1 ml中含胆红素10 μg）。
Take 10 mg of bilirubin control, weigh it precisely, put it into 100 ml brown measuring flask, dissolve and dilute it to the scale with trichloromethane, shake well, measure 5 ml precisely, put it into 50 ml brown measuring flask, add ethanol to the scale, shake well, that is to say, it is obtained (each 1 ml contains 10 μg of bilirubin).
标准曲线的制备 Preparation of standard curves
精密量取对照品溶液1 ml、2 ml、3 ml、4 ml、5 ml，分别置具塞试管中，加乙醇至9 ml，各精密加入重氮化溶液（甲液：取对氨基苯磺酸0. 1 g，加盐酸，1.5 ml与水适量使成100 ml。乙液：取亚硝酸钠0.5 g，用水溶解并稀释至100 ml，置冰箱内保存。临用时取甲液10 ml与乙液0.3 ml，混匀）1 ml,摇匀，在15～20°C的暗处放置1小时，以相应的试剂为空白，照紫外-可见分光光度法（通则0401），在533 nm波长处测定吸光度，以吸光度为纵坐标、浓度为横坐标，绘制标准曲线。
Precisely measure 1 ml, 2 ml, 3 ml, 4 ml, 5 ml of control solution, respectively, placed in a stoppered test tube, add ethanol to 9 ml, add diazotisation solution (solution A: take 0.1 g of p-aminobenzenesulphonic acid, add hydrochloric acid, 1.5 ml and water to make 100 ml; solution B: take 0.5 g of sodium nitrite, dissolve with water and dilute to 100 ml, keep in the refrigerator. At the time of use, take 10 ml of A solution and 0.3 ml of B solution, mix well) 1 ml, shake well, place in the dark at 15-20°C for 1 hour, use the corresponding reagent as the blank, according to the UV-visible spectrophotometric method (General rule 0401), determine the absorbance at the wavelength of 533 nm, take the absorbance as the vertical coordinate, the concentration as the horizontal coordinate, draw the standard curve.
测定法 Method of determination
取本品细粉约10 mg，精密称定，置锥形瓶中，加三氯甲烷-乙醇（7：3）的混合溶液60 ml、盐酸1滴，摇匀，置水浴上加热回流30分钟，放冷，转移至100 ml棕色量瓶中，容器用少量上述混合溶液洗涤，洗液并入同一量瓶中，加上述混合溶液至刻度，摇匀。精密量取上清液10 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀。精密量取3 ml，置具塞试管中，照标准曲线的制备项下的方法，自“加乙醇至9 ml”起，依法测定吸光度，从标准曲线上读岀供试品溶液中含胆红素的重量（mg），计算，即得。
Take about 10 mg of this product fine powder, weighed precisely, placed in a conical flask, add 60 ml of trichloromethane-ethanol (7:3) mixed solution, hydrochloric acid 1 drop, shaking well, placed in a water bath heated reflux for 30 minutes, cooled, and transferred to a 100 ml brown measuring flask, the container was washed with a small amount of the above mixture of solutions, washings were incorporated into the same measuring flask, add the above mixture of solutions to the graduated scale, shaking well. Precisely measure 10 ml of supernatant, put in 50 ml brown measuring flask, add ethanol to the scale, shake well. Precisely measure 3 ml, put in a stoppered test tube, according to the method under the preparation of the standard curve, since "add ethanol to 9 ml", according to the law to determine the absorbance, from the standard curve read out the weight of bilirubin in the test solution (mg), calculated, that is, obtained.
本品按干燥品计算，含胆红素（C33 H36 N4O6）不得少于35.0%。
This product contains not less than 35.0% of bilirubin (C33 H36 N4O6) calculated as dried product.
性 Property
凉。
Cold.
毒性 Toxicity
无。
None.
味 Flavor
甘。
Sweet.
归经 Meridian tropism
心、肝经。
Heart and liver meridians.
功能 Actions
清心，豁痰，开窍，凉肝，息风，解毒。
To clear the heart, resolve phlegm, open the orifices, cool the liver, relieve wind, and detoxify.
主治 Indications
用于热病神昏，中风痰迷，惊痫抽搐，癫痫发狂，咽喉肿痛，口舌生疮，痈肿疔疮。
Used for coma due to febrile diseases, stroke with phlegm obstruction, convulsions of epilepsy, mania of epilepsy, sore throat, oral ulcers, carbuncles and furuncles.
用量 Dosage
0.15～0.35 g。
0.15-0.35 g.
用法 Administration
多入丸散用。外用适量，研末敷患处。
taken in pill or powder form. For external use, apply an appropriate amount of powdered medicine to the affected area.
注意 Precautions
孕妇慎用；偶有轻度消化道不适。
Caution for pregnant women; occasional mild gastrointestinal discomfort.
贮藏 Storage
密闭，遮光，防潮，防压，室温保存。
Store in a sealed, light-proof, moisture-proof, and pressure-resistant container at room temperature.
附：
去氧胆酸质量标准 Deoxycholic acid quality standard
去氧胆酸 Deoxycholic acid
本品由牛胆汁经提取、加工制成。
This product is extracted and processed from bovine bile.
性状 Description
本品为白色的结晶性粉末。气微，味微苦。
This product is a white crystalline powder. It has a slight odor and a slightly bitter taste.
本品易溶于冰醋酸和乙醇，不溶于水。
This product is soluble in glacial acetic acid and ethanol, insoluble in water.
鉴别 Identification
取本品10 mg，加乙醇5 ml使溶解，作为供试品溶液。另取去氧胆酸对照品，加乙醇制成每1 ml含2 mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各2 μl，分别点于同一硅胶G薄层板上，以异辛烷-乙酸乙酯-冰醋酸（15：7：5）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，在105°C加热至斑点显色清晰，置紫外光灯（365 nm）下检视。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的荧光斑点。
Take 10 mg of the product, dissolve it in 5 ml of ethanol, and use it as the test solution. Take an appropriate amount of deoxycholic acid reference substance, dissolve it in ethanol to make a solution containing 2 mg per 1 ml, and use it as the reference solution. Perform a thin-layer chromatography test according to the method specified in <0502>, take 2 μl of each of the above-mentioned solutions, spot them on the same silica gel G thin-layer plate, use n-hexane-ethyl acetate-glacial acetic acid (15:7:5) as the developing agent, develop, take out, dry, spray with a 10% sulfuric acid-ethanol solution, heat at 105°C until the spots are clearly visible, and examine under ultraviolet light (365nm). In the chromatogram of the test solution, there should be fluorescent spots of the same color at the corresponding positions as in the chromatogram of the reference solution.
检查 Examination
干燥失重 Loss on drying
取本品，在105°C干燥4小时，减失重量不得过0.5%（通则0831）。
Take the product, dry it at 105°C for 4 hours, and the loss on drying should not exceed 0.5% <0831>.
炽灼残渣 Residue on ignition
不得过0.2%（通则0841）。
The residue on ignition should not exceed 0.2% <0841>.
含量测定 Content determination
取本品约0. 5 g，精密称定，加中性乙醇（对酚酞指示液显中性）60 ml，置水浴中加热使溶解，冷却，加酚酞指示液数滴及新沸过的冷水20 ml，用氢氧化钠滴定液（0.1 mol/L）滴定，近终点时加新沸过的冷水100 ml，继续滴定至终点。每1ml氢氧化钠滴定液（0.1 mol/L）相当于39. 26 mg 的去氧胆酸（C24 H40O4）。
Take about 0.5g of the product, accurately weigh, place it in a 250 ml conical flask, add 20 ml of water and 40 ml of neutral ethanol (the phenolphthalein indicator solution shows neutrality), heat on a water bath to dissolve, cool, add a few drops of phenolphthalein indicator solution and 20 ml of freshly boiled and cooled water, titrate with sodium hydroxide titration solution (0.1 mol/L) until near the end point, add 100 ml of freshly boiled and cooled water, and continue titration to the end point. Each 1ml of sodium hydroxide titration solution (0.1 mol/L) is equivalent to 39.26 mg of deoxycholic acid (C24H40O4).
本品按干燥品计算，含去氧胆酸（C24 H40O4）不得少于95.0%。
Calculated on the dried basis, the content of deoxycholic acid (C24H40O4) should not be less than 95.0%.
用途 Usage
体外培育牛黄的原料。
Raw material for in vitro cultivation of oxalis.
贮藏 Storage
密封。
Sealing.
胆酸质量标准 Cholic acid quality standard
胆 酸 Cholic acid
本品由牛、羊胆汁或胆膏经提取、加工制成。
This product is extracted and processed from bovine or sheep bile or bile paste.
性状 Description
本品为白色的结晶性粉末。气微，味苦。
This product is a white crystalline powder. It has a slight odor and a bitter taste.
鉴别 Identification
取本品10 mg，加乙醇5 ml使溶解，作为供试品溶液。另取胆酸对照品，加乙醇制成每1 ml含2 mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各2 μl，分别点于同一硅胶G薄层板上，以异辛烷-乙酸乙酯-冰醋酸（15：7：5）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，在105°C加热至斑点显色清晰，置紫外光灯（365nm）下检视。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的荧光斑点。
Take 10 mg of the product, dissolve it in 5 ml of ethanol, and use it as the test solution. Take an appropriate amount of cholic acid reference substance, dissolve it in ethanol to make a solution containing 2 mg per 1 ml, and use it as the reference solution. Perform a thin-layer chromatography test according to the method specified in <0502>, take 2 μl of each of the above-mentioned solutions, spot them on the same silica gel G thin-layer plate, use n-hexane-ethyl acetate-glacial acetic acid (15:7:5) as the developing agent, develop, take out, and dry. Spray with 10% ethanol sulphate solution, heat at 105°C until the spots show clear colour, and examine under UV light (365 nm). In the chromatogram of the test product, in the corresponding position with the chromatogram of the control product, the fluorescent spots of the same colour are shown.
检查 Examination
干燥失重 Loss on drying
取本品，在105°C干燥4小时，减失重量不得过0.5%（通则0831）。
Take the product, dry it at 105°C for 4 hours, and the loss on drying should not exceed 0.5% <0831>.
炽灼残渣 Residue on ignition
不得过0.2%（通则0841）。
The residue on ignition should not exceed 0.2% <0841>.
含量测定 Content determination
取本品约0.4 g，精密称定，置250 ml锥形瓶中，加水20 ml和乙醇40 ml，用表面皿覆盖，置水浴中缓缓加热使完全溶解，冷却，加酚酞指示液5滴，用氢氧化钠滴定液（0. 1 mol/L）滴定。每1 ml氢氧化钠滴定液（0.1 mol/L）相当于40.86 mg的胆酸（C24H40O24）。
Take about 0.4 g of the product, accurately weigh, place it in a 250 ml conical flask, add 20 ml of water and 40 ml of ethanol, cover with a watch glass, heat slowly on a water bath to completely dissolve, cool, add 5 drops of phenolphthalein indicator solution, and titrate with sodium hydroxide titration solution (0.1 mol/L). Each 1 ml of sodium hydroxide titration solution (0.1 mol/L) is equivalent to 40.86 mg of cholic acid (C24H40O24).
本品按干燥品计算，含胆酸（C24H40O24）不得少于95.0%。
Calculated on the dried basis, the content of cholic acid (C24H40O24) should not be less than 95.0%.
用途 Usage
体外培育牛黄的原料。
Raw material for in vitro cultivation of oxalis.
贮藏 Storage
密封。
Sealing.
复合胆红素钙质量标准 Compound Bilirubin Calcium quality standard
复合胆红素钙 Compound Bilirubin Calcium
本品由牛胆汁、胆红素和饱和氢氧化钙溶液经加工制成。
This product is processed from bovine bile, bilirubin, and saturated calcium hydroxide solution.
性状 Description
本品为棕红色或棕黄色的粉末。味微腥。
This product is a brownish-red or brownish-yellow powder. It has a slightly fishy taste.
鉴别 Identification
取本品粉末5 mg，加三氯甲烷15 ml，加盐酸溶液（5→10）0. 1 ml，超声处理30分钟，加三氯甲烷35 ml，摇匀，滤过，取续滤液作为供试品溶液。另取胆红素对照品，加三氯甲烷制成每1 ml含0.1 mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述两种溶液各10 μl，分别点于同一硅胶G薄层板上，以甲苯-乙酸乙酯-冰醋酸（10：1：0.5）为展开剂，展开，取出，晾干。供试品色谱中，在与对照品色谱相应的位置上，显相同颜色的斑点。
Take 5 mg of this product powder, add 15 ml of trichloromethane, add hydrochloric acid solution (5→10) 0.1 ml, ultrasonic treatment for 30 minutes, add 35 ml of trichloromethane, shaking well, filtration, take the filtrate as the test solution. Take bilirubin control product, add trichloromethane to make a solution containing 0.1 mg per 1 ml, as the control solution. According to the thin-layer chromatography (General rule 0502) test, absorb the above two solutions of 10 μl, were spotted on the same silica gel G thin-layer plate, toluene - ethyl acetate - glacial acetic acid (10: 1: 0.5) as an unfolding agent, unfolding, remove, and dry. In the chromatogram of the test article, spots of the same colour are shown in the corresponding positions with the chromatogram of the control article.
检查 Examination
水分 Water
不得过5.0%（通则0832第二法）。
Not more than 5.0 per cent (General Law 0832, second law).
含量测定 Content determination
对照品溶液的制备 Preparation of control solution
取胆红素对照品 10 mg，精密称定，置100 ml棕色量瓶中，用三氯甲烷溶解并稀释至刻度，摇匀。精密量取5 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀，即得（每1 ml中含胆红素10 μg）。
Take 10 mg of bilirubin control product, weigh it precisely, put it in 100 ml brown measuring flask, dissolve and dilute it with trichloromethane to the scale, shake well. Measure 5 ml, put in 50ml brown measuring flask, add ethanol to the scale, shake well, it is obtained (each 1 ml contains 10 μg of bilirubin).
标准曲线的制备 Preparation of standard curves
精密量取对照品溶液1 ml、2 ml、3 ml、4 ml、5 ml，分别置具塞试管中，加乙醇至9 ml，各精密加入重氮化溶液（甲液：取对氨基苯磺酸0.1 g，加盐酸1.5 ml与水适量使成100 ml。乙液：取亚硝酸钠0.5 g，用水溶解并稀释至100 ml，置冰箱内保存。临用时，取甲液10 ml与乙液0.3 ml，混匀）1 ml，摇匀，在15～20°C的暗处放置1小时，以相应的试，剂为空白，照紫外-可见分光光度法（通则0401），在533 nm波长处测定吸光度，以吸光度为纵坐标、浓度为横坐标，绘制标准曲线。
Precisely measure 1 ml, 2 ml, 3 ml, 4 ml, 5 ml of the control solution, respectively, placed in a stoppered test tube, add ethanol to 9 ml, add diazotisation solution (solution A: take 0.1 g of p-aminobenzenesulphonic acid, add 1.5 ml of hydrochloric acid and the appropriate amount of water to make 100 ml; solution B: take 0.5 g of sodium nitrite, dissolve with water and dilute to 100 ml, stored in the refrigerator. At the time of use, take 10 ml of solution A and 0.3 ml of solution B, mix well) 1 ml, shake well, and leave it in the dark at 15-20°C for 1 hour, take the corresponding test agent as blank, according to the UV-visible spectrophotometric method (General rule 0401), determine the absorbance at the wavelength of 533 nm, and plot a standard curve with the absorbance as the vertical coordinates and the concentration as the horizontal coordinates.
测定法 Method of determination
取本品细粉，约10 mg，精密称定，置锥形瓶中，加三氯甲烷-乙醇（7 ：3）的混合溶液60 ml、盐酸1滴，摇匀，置水浴上加热回流30分钟，放冷，转移至100 ml棕色量瓶中，容器用少量上述混合溶液洗涤，洗液并入同一量瓶中，加上述混合溶液至刻度，摇匀。精密量取上清液10 ml，置50 ml棕色量瓶中，加乙醇至刻度，摇匀。精密量取3 ml，置具塞试管中，照标准曲线的制备项下的方法，自“加乙醇至9 ml”起，依法测定吸光度，从标准曲线上读出供试品溶液中含胆红素的重量（mg）,计算，即得。
Take the fine powder of this product, about 10 mg, weighed accurately, placed in a conical flask, add 60 ml of trichloromethane-ethanol (7:3) mixed solution, hydrochloric acid 1 drop, shake well, placed in a water bath heated reflux for 30 minutes, cooled, and transferred to a 100 ml brown measuring flask, the container was washed with a small amount of the above mixture of solutions, washings were incorporated into the same measuring flask, plus the above mixture of solutions to the graduated scale, shaking well. Precisely measure 10 ml of supernatant, put in 50 ml brown measuring flask, add ethanol to the scale, shake well. Precisely measure 3 ml, put in a stoppered test tube, according to the method under the preparation of the standard curve, since "add ethanol to 9 ml", according to the law to determine the absorbance, from the standard curve to read out the weight of bilirubin in the test solution (mg), calculation, that is, obtained.
本品按干燥品计算，含胆红素（C33H36 N4O6）不得少于43.0%。
This product contains not less than 43.0% of bilirubin (C33H36 N4O6) calculated as dry product.
用途 Usage
体外培育牛黄的原料。
Raw material for in vitro cultivation of oxalis.
贮藏 Storage
密封。
Sealing.
本天然药材以牛科动物牛Bos taurus domesticus Gmelin 的新鲜胆汁作母液，加入去氧胆酸、胆酸、复合胆红素钙等制成。
This product is made from the fresh bile of the bovine animal Bos taurus domesticus Gmelin as the base solution, with the addition of deoxycholic acid, cholic acid, and compound bilirubin calcium, etc.