石斛

中文文本参考：《中国药典（2020年版）》
English text reference: Chinese Pharmacopoeia (2020 Edition)

概述 Overview

本品为兰科植物金钗石斛Dendrobium nobile Lindl.、霍山石斛Dendrobium huoshanense C.Z.Tang et S.J.Cheng、鼓槌石斛Dendrobium chrysotoxum Lindl.或流苏石斛Dendrobium fimbriatum Hook.的栽培品及其同属植物近似种的新鲜或干燥茎。全年均可采收，鲜用者除去根和泥沙；干用者采收后，除去杂质，用开水略烫或烘软，再边搓边烘晒，至叶鞘搓净，干燥。霍山石斛11月至翌年3月采收，除去叶、根须及泥沙等杂质，洗净，鲜用，或加热除去叶鞘制成干条；或边加热边扭成螺旋状或弹簧状，干燥，称霍山石斛枫斗。
Dendrobium Stem is the dried stem of cultivated Dendrobium nobile Lindl., Dendrobium huoshanense C.Z.Tang et S.J.Cheng, Dendrobium chrysotoxum Lindl. or Dendrobium fimbriatum Hook. (Fam. Orchidaceae), or their similar species. It can be collected throughout the year. For fresh use, remove the roots and impurities. For dried use, remove the impurities after collection, scald or soften with boiling water, rub and dry until the leaf sheaths are clean and dry. Dendrobium huoshanense is collected from November to March of the following year. Remove the leaves, rootlets, and impurities, wash, and use fresh, or heat to remove the leaf sheaths and make dried strips, or heat and twist into spiral or spring shapes, and dry, called Fengdou.

性状 Description

鲜石斛 呈圆柱形或扁圆柱形，长约30cm，直径0.4～1.2cm。表面黄绿色，光滑或有纵纹，节明显，色较深，节上有膜质叶鞘。肉质多汁，易折断。气微，味微苦而回甜，嚼之有黏性。
Fresh Dendrobium Stem is cylindrical or slightly flattened cylindrical, about 30 cm long, with a diameter of 0.4-1.2 cm. The surface is yellowish-green, smooth or with longitudinal lines, with obvious nodes and darker color, and membranous leaf sheaths on the nodes. The flesh is juicy and easily broken. It has a slight odor, slightly bitter and sweet taste, and chewy texture.

金钗石斛 呈扁圆柱形，长20～40cm，直径0.4～0.6cm，节间长2.5～3cm。表面金黄色或黄中带绿色，有深纵沟。质硬而脆，断面较平坦而疏松。气微，味苦。
Dendrobium nobile is flattened cylindrical, 20-40 cm long, 0.4-0.6 cm in diameter, with internodes 2.5-3 cm long. The surface is golden yellow or yellowish-green with deep longitudinal grooves. The texture is hard and brittle, and the fracture surface is relatively flat and loose. It has a slight odor and a bitter taste.

霍山石斛 干条呈直条状或不规则弯曲形，长2～8cm，直径1～4mm。表面淡黄绿色至黄绿色，偶有黄褐色斑块，有细纵纹，节明显，节上有的可见残留的灰白色膜质叶鞘；一端可见茎基部残留的短须根或须根痕，另一端为茎尖，较细。质硬而脆，易折断，断面平坦，灰黄色至灰绿色，略角质状。气微，味淡，嚼之有黏性。鲜品稍肥大。肉质，易折断，断面淡黄绿色至深绿色。气微，味淡，嚼之有黏性且少有渣。枫斗呈螺旋形或弹簧状，通常为2～5个旋纹，茎拉直后性状同干条。
Dendrobium huoshanense is dried strips that are straight or irregularly curved, 2-8 cm long, and 1-4 mm in diameter. The surface is light yellowish-green to yellowish-green, occasionally with yellow-brown patches, fine longitudinal lines, obvious nodes, and residual grayish-white membranous leaf sheaths on the nodes. One end may have short rootlets or rootlet scars remaining at the base of the stem, and the other end is the stem tip, which is thinner. The texture is hard and brittle, easily broken, with a flat fracture surface, grayish-yellow to grayish-green, and slightly horny. It has a slight odor, a mild taste, and chewiness. The fresh stem is slightly fleshy and larger. The flesh is easily broken, and the fracture surface is light yellowish-green to dark green. It has a slight odor, a mild taste, chewiness, and few impurities. Fengdou is spiral or spring-shaped, usually with 2-5 spirals, and has the same characteristics as dried strips when the stem is straightened.

鼓槌石斛 呈粗纺锤形，中部直径1～3cm，具3～7节。表面光滑，金黄色，有明显凸起的棱。质轻而松脆，断面海绵状。气微，味淡，嚼之有黏性。
Dendrobium chrysotoxum is coarse fusiform, with a diameter of 1-3 cm in the middle and 3-7 nodes. The surface is smooth, golden yellow, with obvious raised edges. The texture is light and brittle, and the fracture surface is spongy. It has a slight odor, a mild taste, and chewiness.

流苏石斛等 呈长圆柱形，长20～150cm，直径0.4～1.2cm，节明显，节间长2～6cm。表面黄色至暗黄色，有深纵槽。质疏松，断面平坦或呈纤维性。味淡或微苦，嚼之有黏性。
Dendrobium fimbriatum and others are long cylindrical, 20-150 cm long, 0.4-1.2 cm in diameter, with obvious nodes and internodes 2-6 cm long. The surface is yellow to dark yellow with deep longitudinal grooves. The texture is loose, and the fracture surface is flat or fibrous. It has a mild or slightly bitter taste and chewiness.

鉴别 Identification

（1）本品横切面：金钗石斛 表皮细胞1列，扁平，外被鲜黄色角质层。基本组织细胞大小较悬殊，有壁孔，散在多数外韧型维管束，排成7～8圈。维管束外侧纤维束新月形或半圆形，其外侧薄壁细胞有的含类圆形硅质块，木质部有1～3个导管直径较大。含草酸钙针晶细胞多见于维管束旁。
Dendrobium nobile transverse section: Epidermal cells are in one layer, flat, and covered with a bright yellow horny layer. The basic tissue cells vary greatly in size, have pit canals, and are scattered in most external collateral vascular bundles, arranged in 7-8 circles. The outer fiber bundles of the vascular bundle are crescent-shaped or semicircular, and the outer thin-walled cells sometimes contain circular siliceous bodies. The xylem has 1-3 larger diameter vessels. Calcium oxalate needle crystals are common in the vicinity of the vascular bundle.

霍山石斛 表皮细胞1列，扁平，外壁及侧壁稍增厚，微木化，外被黄色或橘黄色角质层，有的外层可见无色的薄壁细胞组成的叶鞘层。基本薄壁组织细胞多角形，大小相似，其间散在9～47个维管束，近维管束处薄壁细胞较小，维管束为有限外韧型，维管束鞘纤维群呈单帽状，偶成双帽状，纤维1～2列，外侧纤维直径通常小于内侧纤维，有的外侧小型薄壁细胞中含有硅质块。草酸钙针晶束多见于近表皮处薄壁细胞或近表皮处维管束旁的薄壁细胞中。
Dendrobium huoshanense: Epidermal cells are in one layer, flat, with slightly thickened outer and lateral walls, slightly lignified, and covered with a yellow or orange horny layer. Some outer layers are composed of colorless thin-walled cells forming a leaf sheath layer. The basic thin-walled tissue cells are polygonal and similar in size. There are 9-47 vascular bundles scattered among them. The thin-walled cells near the vascular bundles are smaller. The vascular bundles are externally collateral and limited. The fiber groups of the vascular bundle sheath are single or double-capped, with 1-2 rows of fibers. The diameter of the outer fibers is usually smaller than that of the inner fibers. Some small thin-walled cells in the outer layer contain siliceous bodies. Calcium oxalate needle crystal bundles are common in the thin-walled cells near the epidermis or near the vascular bundles.

鼓槌石斛 表皮细胞扁平，外壁及侧壁增厚，胞腔狭长形；角质层淡黄色。基本组织细胞大小差异较显著。多数外韧型维管束略排成10～12圈。木质部导管大小近似。有的可见含草酸钙针晶束细胞。
Dendrobium chrysotoxum: Epidermal cells are flat, with thickened outer and lateral walls and narrow elongated cell cavities; the horny layer is light yellow. There are significant differences in the size of the basic tissue cells. Most external collateral vascular bundles are arranged in 10-12 circles. The size of the xylem vessels is similar. Some cells containing calcium oxalate needle crystal bundles can be seen.

流苏石斛等 表皮细胞扁圆形或类方形，壁增厚或不增厚。基本组织细胞大小相近或有差异，散列多数外韧型维管束，略排成数圈。维管束外侧纤维束新月形或呈帽状，其外缘小细胞有的含硅质块；内侧纤维束无或有，有的内外侧纤维束连接成鞘。有的薄壁细胞中含草酸钙针晶束和淀粉粒。
Dendrobium fimbriatum and others: Epidermal cells are flattened round or square, with thickened or non-thickened walls. The size of the basic tissue cells is similar or different, scattered in most external collateral vascular bundles, arranged in several circles. The outer fiber bundles of the vascular bundle are crescent-shaped or cap-shaped, and the outer edge of the small cells contains siliceous bodies; the inner fiber bundles are absent or present, and some inner and outer fiber bundles are connected to form a sheath. Some thin-walled cells contain calcium oxalate needle crystal bundles and starch grains.

粉末灰绿色或灰黄色。角质层碎片黄色；表皮细胞表面观呈长多角形或类多角形，垂周壁连珠状增厚。束鞘纤维成束或离散，长梭形或细长，壁较厚，纹孔稀少，周围具排成纵行的含硅质块的小细胞。木纤维细长，末端尖或钝圆，壁稍厚。网纹导管、梯纹导管或具缘纹孔导管直径12～50μm。草酸钙针晶成束或散在。
The powder is grayish-green or grayish-yellow. The fragments of the horny layer are yellow. The surface of the epidermal cells is elongated polygonal or polygonal, and the radial walls are thickened in a beaded pattern. The fiber sheaths are bundled or scattered, elongated or slender, with thicker walls, few pits, and small cells with siliceous bodies arranged in longitudinal rows around them. The wood fibers are elongated, with pointed or blunt ends and slightly thicker walls. The reticulate vessels, scalariform vessels, or vessels with bordered pits have a diameter of 12-50 μm. Calcium oxalate needle crystals are bundled or scattered.

（2）金钗石斛 取本品（鲜品干燥后粉碎）粉末1g，加甲醇10ml，超声处理30分钟，滤过，滤液作为供试品溶液。另取石斛碱对照品，加甲醇制成每1ml含1mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取供试品溶液20μl、对照品溶液5μl，分别点于同一硅胶G薄层板上，以石油醚（60～90℃）-丙酮（7:3）为展开剂，展开，取出，晾干，喷以碘化铋钾试液。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。
Dendrobium nobile: Take 1g of the powder (after drying the fresh product), add 10ml of methanol, and sonicate for 30 minutes. Filter and use the filtrate as the test solution. Take Dendrobine CRS, add methanol to make a solution containing 1mg per ml, and use it as the reference solution. Perform thin-layer chromatography<0502> using the test solution (20μl) and the reference solution (5μl) on the same silica gel G plate, with petroleum ether (60-90°C)-acetone (7:3) as the mobile phase. Develop, remove, dry, and spray with a solution of bismuth potassium iodide. In the chromatogram of the test solution, there should be spots of the same color at the corresponding positions as in the chromatogram of the reference solution.

霍山石斛 取本品（鲜品干燥后粉碎）粉末（过二号筛）1g，加无水甲醇20ml，超声处理30分钟，滤过，滤液回收溶剂至干，残渣加水15ml使溶解，用石油醚（60～90℃）洗涤2次，每次20ml，弃去石油醚液，水液用乙酸乙酯洗涤2次，每次20ml，弃去乙酸乙酯液，用水饱和正丁醇振摇提取2次，每次20ml，合并正丁醇液，回收溶剂至干，残渣加无水甲醇1ml使溶解，作为供试品溶液。另取霍山石斛对照药材1g，同法制成对照药材溶液。再取夏佛塔苷对照品适量，加甲醇制成每1ml含0.5mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述三种溶液各3～5μl，分别点于同一聚酰胺薄膜上，以乙醇-丁酮-乙酰丙酮-水（4:4:1:17）为展开剂，20℃以下展开，取出，晾干，在105℃烘干，取出，喷以5%三氯化铝乙醇溶液，在105℃加热约3分钟，取出，置紫外光灯（365nm）下检视。供试品色谱中，在与对照药材色谱和对照品色谱相应的位置上，显相同颜色的荧光斑点。
Dendrobium huoshanense: Take 1g of the powder (after drying the fresh product, passed through a No. 2 sieve), add 20ml of anhydrous methanol, sonicate for 30 minutes, filter, and recover the solvent from the filtrate to dryness. Dissolve the residue in 15ml of water, wash with petroleum ether (60-90°C) twice, each time with 20ml, discard the petroleum ether solution, wash the aqueous solution with ethyl acetate twice, each time with 20ml, discard the ethyl acetate solution, extract with saturated n-butanol twice, each time with 20ml, combine the n-butanol solution, recover the solvent to dryness, and dissolve the residue in 1ml of anhydrous methanol as the test solution. Take 1g of Dendrobium huoshanense as the reference material, and prepare the reference material solution using the same method. Take an appropriate amount of Forsythoside A CRS, add methanol to make a solution containing 0.5mg per ml as the reference solution. Perform thin-layer chromatography<0502> using 3-5μl of each of the above three solutions on the same polyamide film, with ethanol-butanone-acetylacetone-water (4:4:1:17) as the mobile phase, develop below 20°C, remove, dry, dry at 105°C, remove, and spray with a 5% solution of aluminum chloride in ethanol, heat at 105°C for about 3 minutes, remove, and examine under ultraviolet light at 365 nm. In the chromatogram of the test solution, there should be fluorescent spots of the same color at the corresponding positions as in the chromatograms of the reference material and the reference solution.

鼓槌石斛 取鼓槌石斛〔含量测定〕项下的续滤液25ml，回收溶剂至干，残渣加甲醇5ml使溶解，作为供试品溶液。另取毛兰素对照品，加甲醇制成每1ml含0.2mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取供试品溶液5～10μl、对照品溶液5μl，分别点于同一高效硅胶G薄层板上，以石油醚（60～90℃）-乙酸乙酯（3:2）为展开剂，展开，展距8cm，取出，晾干，喷以10%硫酸乙醇溶液，在105℃加热至斑点显色清晰。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。
Dendrobium chrysotoxum: Take 25ml of the subsequent filtrate under the "Determination of Content" section of Dendrobium chrysotoxum, recover the solvent to dryness, dissolve the residue in 5ml of methanol, and use it as the test solution. Take Resveratrol CRS, add methanol to make a solution containing 0.2mg per ml, and use it as the reference solution. Perform thin-layer chromatography<0502> using 5-10μl of the test solution and 5μl of the reference solution on the same high-performance silica gel G plate, with petroleum ether (60-90°C)-ethyl acetate (3:2) as the mobile phase. Develop, remove, dry, and spray with a 10% solution of sulfuric acid in ethanol, heat at 105°C until the spots are clearly visible. In the chromatogram of the test solution, there should be spots of the same color at the corresponding positions as in the chromatogram of the reference solution.

流苏石斛等 取本品（鲜品干燥后粉碎）粉末0.5g，加甲醇25ml，超声处理45分钟，滤过，滤液蒸干，残渣加甲醇5ml使溶解，作为供试品溶液。另取石斛酚对照品，加甲醇制成每1ml含0.2mg的溶液，作为对照品溶液。照薄层色谱法（通则0502）试验，吸取上述供试品溶液5～10μl、对照品溶液5μl，分别点于同一高效硅胶G薄层板上，以石油醚（60～90℃）-乙酸乙酯（3:2）为展开剂，展开，展距8cm，取出，晾干，喷以10%硫酸乙醇溶液，在105℃加热至斑点显色清晰。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。
Dendrobium fimbriatum and others：Take 0.5g of this product (fresh and crushed after drying) powder, add 25ml of methanol, ultrasonic treatment for 45 minutes, filtration, filtrate evaporation, residue add 5ml of methanol to make dissolved, as a test solution. Another take dendrobenol control, add methanol to make a solution containing 0.2mg per 1ml, as a control solution. According to the thin layer chromatography (General rule 0502) test, absorb the above test solution 5 ~ 10 μl, control solution 5 μl, were spotted on the same high-performance silica gel G thin layer plate, with petroleum ether (60 ~ 90 ℃) - ethyl acetate (3:2) as the unfolding agent, unfolding, spreading distance of 8cm, remove, drying, sprayed with 10% sulfuric acid ethanol solution, heated to 105 ℃ until the spots show clear colour. In the chromatogram of the test article, in the corresponding position with the chromatogram of the control article, the spot of the same colour is shown.

（3）霍山石斛 聚合酶链式反应-限制性内切酶长度多态性方法。
(3) Dendrobium Huoshan Polymerase chain reaction-restriction endonuclease length polymorphism method.

　 模板DNA提取 取本品0.1g（鲜品干燥），加液氮适量研磨，过五号筛。取粉末25mg，置1.5ml离心管中，加入CTAB沉淀液［2%十六烷基三甲基溴化铵，100mmol/L Tris-盐酸pH=8.0，10mmol/L乙二胺四乙酸二钠］1000μl，涡旋震荡，65℃水浴加热20分钟（中间震荡混匀3次），离心（转速为每分钟12000转）10分钟，弃去上清液；再加入CTAB沉淀液1000μl，涡旋震荡，65℃水浴加热10分钟，离心（转速为每分钟12000转）10分钟，弃去上清液；再同法操作一次；弃去上清液，加入CTAB提取液［2%十六烷基三甲基溴化铵，100mmol/L Tris-盐酸pH=8.0，20mmol/L乙二胺四乙酸二钠，2.5mol/L氯化钠］900μl、蛋白酶K（20mg/ml）5μl充分混匀，65℃水浴加热30分钟，离心（转速为每分钟12000转）10分钟，吸取上清液置另一2.0ml离心管中；加入900μl三氯甲烷-异戊醇（体积比24：1）溶液，充分混匀，离心（转速为每分钟12000转）10分钟；取上清液，加入等体积三氯甲烷-异戊醇（体积比24：1）溶液（约800μl），充分混匀，离心（转速为每分钟12000转）10分钟；取上清液置另一2.0ml离心管中，加入2/3体积的异丙醇，置-20℃放置30分钟；离心（转速为每分钟12000转）10分钟，弃去上清液；沉淀加70%乙醇500μl震荡1分钟，离心（转速为每分钟12000转）3分钟；弃去上清液，沉淀再用70%乙醇500μl震荡1分钟，离心（转速为每分钟12000转）3分钟；弃去上清液，置37℃水浴中挥干溶剂；加入高压灭菌超纯水50μl，溶解，作为供试品溶液，置4℃冰箱中备用。另取霍山石斛对照药材0.1g，同法制成对照药材模板DNA溶液。
Template DNA extraction Take 0.1g of this product (fresh and dried), add appropriate amount of liquid nitrogen and grind it, and pass it through sieve number five. Take 25mg of powder, put in 1.5ml centrifuge tube, add CTAB precipitation solution [2% cetyltrimethylammonium bromide, 100mmol/L Tris-hydrochloric acid pH=8.0, 10mmol/L disodium ethylenediaminetetraacetic acid] 1000μl, vortexing and shaking, heating at 65℃ for 20 minutes (with shaking and mixing for 3 times in the middle), centrifugation (rotating speed 12000 rpm) 10 minutes, discard the supernatant; then add CTAB precipitation solution 1000μl, and discard the supernatant; then add CTAB precipitation solution 1000μl. Then add 1000 μl of CTAB precipitation solution, vortex shaking, heating at 65℃ for 10 minutes, centrifugation (speed of 12000 rpm) for 10 minutes, and discard the supernatant; then do the same operation once more; discard the supernatant, and add the CTAB extraction solution [2% cetyltrimethylammonium bromide, 100 mmol/L Tris-hydr ochloric acid pH=8.0, 20 mmol/L Tris-hydrochloric acid], and then add the CTAB extraction solution [2% cetyltrimethylammonium bromide, 100 mmol/L Tris-hydrochloric acid, pH=8.0, 20 mmol/L Tris-hydrochloric acid]. 8.0, 20 mmol/L disodium ethylenediaminetetraacetic acid, 2.5 mol/L sodium chloride] 900 μl, proteinase K (20 mg/ml) 5 μl, mixed thoroughly, 65 ℃ water bath heating for 30 minutes, centrifugation (speed of 12,000 revolutions per minute) for 10 minutes, aspirated supernatant in another 2.0 ml centrifugal tube; add 900 μl of trichloromethane-isoamyl alcohol (v/v) solution, mixed thoroughly. Add 900μl of trichloromethane-isoamyl alcohol (24:1, v/v) solution, mix thoroughly and centrifuge (at 12,000 rpm) for 10 minutes; take the supernatant and add an equal volume of trichloromethane-isoamyl alcohol (24:1, v/v) solution (about 800μl), mix thoroughly and centrifuge (at 12,000 rpm) for 10 minutes; take the supernatant and put it into another 2.0 ml centrifugal tube, add 2/3 volume of isopropanol and lea ve it at -20℃ for 30 minutes; centrifuge (at -20℃ for 30 minutes). The supernatant was placed in another 2.0 ml centrifuge tube, 2/3 volume of isopropanol was added, and placed at -20℃ for 30 min; centrifugation was performed at 12,000 rpm for 10 min, and the supernatant was discarded; the precipitate was added with 500 μl of 70% ethanol and shaken for 1 min, and centrifugation was performed at 12,000 rpm for 3 min; the supernatant was discarded, and the solvent was evaporated in a water bath at 37℃, and placed in a water bath at -30℃. 37 ℃ water bath, evaporate the solvent; add autoclaved ultrapure water 50 μl, dissolve, as a test solution, placed in a refrigerator at 4 ℃ standby. Take 0.1g of Dendrobium huoshanense control herb, and make template DNA solution of control herb by the same method.

PCR-RFLP反应 鉴别引物：5'-ATTCTTCATCAAGTTTAGTGCATTC-3'和5'-AGAGCTGATGGGCCTTTGA-3'。PCR反应体系：在200μl离心管中进行，反应总体积为25μl，反应体系包括10×PCR缓冲液2.5μl，dNTP（10mmol/L）1μl，鉴别引物（10μmol/L）各0.2μl，Taq DNA聚合酶（5U/μl）0.2μl，10mg/ml牛血清蛋白1μl，25%聚乙烯吡咯烷酮1μl，模板1μl，无菌超纯水18.4μl。将离心管置PCR仪，PCR反应参数：95℃预变性5分钟，循环反应40次（95℃ 10秒，56℃ 20秒，72℃ 20秒），72℃延伸5分钟。取PCR反应液，置200μl离心管中，进行酶切反应，反应总体积为20μl，反应体系包括10×酶切缓冲液2μl，PCR反应液17.5μl，AluⅠ内切酶（10U/μl）0.5μl，酶切反应在37℃水浴反应30分钟。另取无菌超纯水，同法上述PCR-RFLP反应操作，作为空白对照。
PCR-RFLP reaction Identifying primers: 5'-ATTCTTCATCAAGTTTAGTGCATTC-3' and 5'-AGAGCTGATGGGGCCTTTGA-3'. PCR reaction system: carried out in 200 μl centrifuge tubes, the total volume of the reaction is 25 μl, the reaction system includes 10 × PCR buffer 2.5 μl, dNTP （The reaction system included 10×PCR buffer 2.5 μl, dNTP (10 mmol/L) 1 μl, identification primers (10 μmol/L) 0.2 μl each, Taq DNA polymerase (5 U/μl) 0.2 μl, 10 mg/ml bovine serum proteins 1 μl, 25% polyvinylpyrrolidone 1 μl, templates 1 μl, and 18.4 μl of sterile ultra-pure water. PCR reaction parameters: pre-denaturation at 95℃ for 5 minutes, cycling reaction for 40 times (95℃ for 10 seconds, 56℃ for 20 seconds, 72℃ for 20 seconds), extension at 72℃ for 5 minutes. The PCR reaction solution was taken and placed in a 200 μl centrifuge tube, and the total volume of the reaction was 20 μl, and the reaction system consisted of 2 μl of 10× digestion buffer, 17.5 μl of PCR reaction solution, and 0.5 μl of Alu Ⅰ endonuclease (10 U/μl), and the digest ion reaction was carried out in a water bath at 37 ℃ for 30 minutes. Another sterile ultrapure water was taken and operated in the same way as the above PCR-RFLP reaction, which was used as a blank control.

电泳检测 照琼脂糖凝胶电泳法（通则0541），胶浓度为2.5%，胶中加入核酸凝胶染色剂GelRed；以DL500作为DNA分子量标记，供试品与对照药材PCR产物和酶切反应产物的上样量分别各8μl，DNA分子量标记上样量为2μl（0.5μg/μl）。电泳结束后，取凝胶片在凝胶成像仪上或紫外透射仪上检视。霍山石斛供试品凝胶电泳图谱中，在与对照药材凝胶电泳图谱相应位置上，在100～200bp间应有单一DNA条带，且PCR产物与酶切产物条带位置一致。空白对照无条带。
Electrophoresis detection According to the agarose gel electrophoresis method (General rule 0541), the concentration of the gel was 2.5%, and GelRed, a nucleic acid gel stain, was added to the gel; DL500 was used as the DNA molecular weight marker, and the sample volume of PCR products and enzyme digestion products of the test and control herbs was 8 μl each, and the sample volume of the DNA molecular weight marker was 2 μl (0.5 μg/μl). After electrophoresis, the gel slices were taken and examined on a gel imager or UV transilluminator. In the gel electrophoresis profile of Dendrobium huoshanense test material, there should be a single DNA band between 100 and 200 bp in the corresponding position of the gel electrophoresis profile of the control herb, and the positions of the bands of the PCR product and the enzyme digestion product were the same. The blank control has no bands.

特征图谱 Atlas of features

霍山石斛 照高效液相色谱法（通则0512测定）。
Dendrobium huoshanensis Determined by high performance liquid chromatography (General rule 0512).

色谱条件与系统适用性试验 以十八烷基硅烷键合硅胶为填充剂；以乙腈-甲醇溶液（1:1）为流动相A，0.01mol/L乙酸铵溶液为流动相B，按下表中的规定进行梯度洗脱；流速为每分钟0.8ml；柱温40℃；检测波长为340nm。理论板数按夏佛塔苷峰计算应不低于5000。
Chromatographic conditions and system suitability test The chromatographic conditions were as follows: Octadecylsilane-bonded silica gel as filler; Acetonitrile-methanol solution (1:1) as mobile phase A, 0.01mol/L ammonium acetate solution as mobile phase B; Gradient elution according to the following table; Flow rate of 0.8 ml per minute; Column temperature of 40 ℃; Detection wavelength of 340 nm; Theoretical plate counts based on the chafferetin peaks should be not less than 5000. The theoretical plate count should be not less than 5000 based on the peak of charophylloside.

参照物溶液的制备 取霍山石斛对照药材约1g，加入甲醇50ml，超声处理30分钟（功率250W，频率50kHz），取出，放冷，滤过，滤液浓缩至5ml，作为对照药材参照物溶液。另取夏佛塔苷对照品加甲醇制成每1ml含50μg的溶液，作为对照品参照物溶液，即得。
Preparation of reference solution Take about 1g of Huoshan Dendrobium control herb, add 50ml of methanol, ultrasonic treatment for 30 minutes (power 250W, frequency 50kHz), take out, cool down, filter, filtrate and concentrate the filtrate to 5ml as the reference solution of control herb. Another take Xaveratoside control with methanol to make a solution containing 50μg per 1ml, as a control reference solution, that is obtained.

供试品溶液的制备 取本品（鲜品干燥后粉碎）粉末（过三号筛）约1g，同对照药材参照物溶液制备方法，制成供试品溶液，即得。
Preparation of test solution Take about 1g of this product (fresh and crushed after drying) powder (through the No.3 sieve), the same as the control herb reference solution preparation method, make a test solution, that is, obtained.

测定法 分别精密吸取上述参照物溶液与供试品溶液各5～20μl，注入液相色谱仪，记录色谱图，即得。
Determination Method Precisely aspirate 5～20μl each of the above reference solution and test solution respectively, inject into liquid chromatograph, record the chromatogram, and it is obtained.

供试品色谱中应呈现5个特征峰，并应与对照药材参照物色谱峰中的5个特征峰保留时间相对应，其中峰1应与对照品参照物峰保留时间相对应。
Five characteristic peaks should be presented in the chromatogram of the test article and should correspond to the retention times of the five characteristic peaks in the chromatogram peak of the reference of the control herb, of which peak 1 should correspond to the retention time of the peak of the reference of the control article.

检查 Examination

水分 Water

干石斛不得过12.0%（通则0832第二法）。
Dendrobium officinale should not be more than 12.0 per cent <0832,method 2>.

总灰分 Total ash

干石斛 不得过5.0%（通则2302）。
Dendrobium officinale should not be more than 5.0 per cent <2302>.

霍山石斛 不得过7.0%（通则2302）。
Houshan Dendrobium should not be more than 7.0 per cent <2302>.

浸出物 Extractives

霍山石斛 照醇溶性浸出物测定法（通则2201）项下的热浸法测定，用乙醇作溶剂，干品不得少于8.0%。
Carry out the method for determination of ethanol-soluble Extractives <2201,the hot maceration method>, using ethanol as the solvent,not less than 8.0 per cent.

含量测定 Content determination

金钗石斛 照气相色谱法（通则0521）测定。
Carry out the method for determination of content by gas chromatography <0521>.

色谱条件与系统适用性试验 Chromatographic conditions and system suitability test

DB-1毛细管柱（100%二甲基聚硅氧烷为固定相）（柱长为30m，内径为0.25mm，膜厚度为0.25μm），程序升温：初始温度为80℃，以每分钟10℃的速率升温至250℃，保持5分钟；进样口温度为250℃，检测器温度为250℃。理论板数按石斛碱峰计算应不低于10000。
DB-1 capillary column (100% dimethylpolysiloxane as stationary phase) (column length of 30 m, inner diameter of 0.25 mm, film thickness of 0.25 μm), programmed temperature increase: the initial temperature was 80 ℃, and the temperature was increased to 250 ℃ at the rate of 10 ℃ per minute, and held for 5 minutes; the temperature of the inlet port was 250 ℃, and the temperature of the detector was 250 ℃. The theoretical plate number should be not less than 10000 based on the dendrobase peak.

校正因子测定 Calibration factor determination

取萘对照品适量，精密称定，加甲醇制成每lml含25μg的溶液，作为内标溶液。取石斛碱对照品适量，精密称定，加甲醇制成每1ml含50μg的溶液，作为对照品溶液。精密量取对照品溶液2ml，置5ml量瓶中，精密加入内标溶液1ml，加甲醇至刻度，摇匀，吸取1μl，注入气相色谱仪，计算校正因子。
Take the appropriate amount of naphthalene control, weighing precision, add methanol to make a solution containing 25μg per lml, as the internal standard solution. Take the appropriate amount of Dendrobine control, precision weighing, add methanol to make a solution containing 50μg per 1ml, as the control solution. Precisely measure 2ml of the control solution, put it in a 5ml measuring flask, add 1ml of the internal standard solution, add methanol to the scale, shake well, take up 1μl, inject it into the gas chromatograph and calculate the correction factor.

测定法 Determination method

取本品（鲜品干燥后粉碎）粉末（过三号筛）约0.25g，精密称定，置圆底烧瓶中，精密加入0.05%甲酸的甲醇溶液25ml，称定重量，加热回流3小时，放冷，再称定重量，用0.05%甲酸的甲醇溶液补足减失的重量，摇匀，滤过。精密量取续滤液2ml，置5ml量瓶中，精密加入内标溶液1ml，加甲醇至刻度，摇匀，吸取1μl，注入气相色谱仪，测定，即得。
Take the product (fresh and crushed after drying) powder (through the third sieve) about 0.25g, weighing, placed in a round-bottomed flask, add 0.05% formic acid methanol solution 25ml, weighing, heating reflux for 3 hours, cooling, then weighing, with 0.05% formic acid methanol solution to make up for the loss of weight, shaking well, filtered. Precisely measure 2ml of filtrate, put it in 5ml measuring flask, add 1ml of internal standard solution, add methanol to the scale, shake well, take up 1μl, inject it into gas chromatograph, and then determine, it is obtained.

本品按干燥品计算，含石斛碱（C16H25NO2）不得少于0.40%。
This product is calculated according to the dry product, containing dendrobine (C16H25NO2) shall not be less than 0.40%.

霍山石斛 照紫外-可见分光光度法（通则0401）测定。
Houshan Dendrobium Carry out the method for determination by ultraviolet-visible spectrophotometry <0401>.

对照品溶液的制备 Preparation of reference solution

取D-无水葡萄糖对照品适量，精密称定，加水制成每1ml含100μg的溶液，即得。
Take an appropriate amount of D-glucose reference substance, accurately weigh, add water to make a solution containing 100μg per ml, as the reference solution.

标准曲线的制备 Preparation of standard curve

精密量取对照品溶液0.2ml、0.4ml、0.6ml、0.8ml、1.0ml，分别置10ml具塞试管中，各加水至1.0ml，精密加入5%苯酚溶液1ml（临用配制），摇匀，再精密加硫酸5ml，摇匀，置沸水浴中加热20分钟，取出，置冰浴中冷却5分钟，以相应试剂为空白，照紫外-可见分光光度法（通则0401），在488nm的波长处测定吸光度，以吸光度为纵坐标，浓度为横坐标，绘制标准曲线。
Accurately take 0.2ml, 0.4ml, 0.6ml, 0.8ml, and 1.0ml of the reference solution, respectively, place them in 10ml stoppered test tubes, add water to 1.0ml each, accurately add 1ml of 5% phenol solution (prepared for immediate use), shake well, then accurately add 5ml of sulfuric acid, shake well, heat in a boiling water bath for 20 minutes, take out, cool in an ice bath for 5 minutes, use the corresponding reagent as a blank, determine the absorbance at a wavelength of 488nm by ultraviolet-visible spectrophotometry <0401>, and plot the standard curve with absorbance as the ordinate and concentration as the abscissa.

测定法 Determination method

取本品（鲜品干燥后粉碎）粉末（过三号筛）约0.4g，精密称定，加水200ml，加热回流2小时，放冷，转移至250ml量瓶中，用少量水分次洗涤容器，洗液并入同一量瓶中，加水至刻度，摇匀，滤过。精密量取续滤液2ml，置15ml离心管中，精密加入无水乙醇10ml，摇匀，冷藏1小时，取出，离心（转速为每分钟4000转）20分钟，弃去上清液，沉淀加80%乙醇洗涤2次，每次8ml，离心，弃去上清液，沉淀加热水溶解，转移至25ml量瓶中，放至室温，加水至刻度，摇匀。精密量取供试品溶液1ml，置10ml具塞试管中，照标准曲线制备项下的方法，自“精密加入5%苯酚溶液1ml”起依法测定吸光度，从标准曲线上读出供试品溶液中D-无水葡萄糖的量，计算，即得。
Take about 0.4g of the powder of the product (dried and crushed) (passed through a No. 3 sieve), accurately weigh it, add 200ml of water, heat under reflux for 2 hours, cool, transfer to a 250ml volumetric flask, wash the container with a small amount of water in portions, and combine the washings into the same volumetric flask, add water to the mark, shake well, and filter. Accurately take 2ml of the filtrate, place it in a 15ml centrifuge tube, accurately add 10ml of anhydrous ethanol, shake well, refrigerate for 1 hour, take out, centrifuge (at a speed of 4000 rpm per minute) for 20 minutes, discard the supernatant, wash the precipitate with 80% ethanol twice, each time with 8ml, discard the supernatant, dissolve the precipitate in hot water, transfer to a 25ml volumetric flask, let stand at room temperature, add water to the mark, shake well. Accurately take 1ml of the test solution, place it in a 10ml stoppered test tube, determine the absorbance according to the method under "Preparation of standard curve", starting from "accurately add 1ml of 5% phenol solution", and read the amount of D-glucose in the test solution from the standard curve, calculate, and obtain the result.

本品按干燥品计算，含多糖以无水葡萄糖（C6H12O6）计，不得少于17.0%。
The product is calculated on the basis of the dried product, and the content of polysaccharides is calculated as anhydrous glucose (C6H12O6), which should not be less than 17.0%.

鼓槌石斛 照高效液相色谱法（通则0512）测定。
Guchui Dendrobium Carry out the method for determination by high performance liquid chromatography <0512>.

色谱条件与系统适用性试验 Chromatographic conditions and system suitability test

以十八烷基硅烷键合硅胶为填充剂；以乙腈-0.05%磷酸溶液（37:63）为流动相；检测波长为230nm。理论板数按毛兰素峰计算应不低于6000。
Use octadecylsilane bonded silica gel as the filler; use acetonitrile-0.05% phosphoric acid solution (37:63) as the mobile phase; detection wavelength 230nm. The theoretical number of plates calculated based on the peak of moscatilin should not be less than 6000.

对照品溶液的制备 Preparation of reference solution

取毛兰素对照品适量，精密称定，加甲醇制成每1ml含15μg的溶液，即得。
Take an appropriate amount of moscatilin reference substance, accurately weigh, add methanol to make a solution containing 15μg per ml, as the reference solution.

供试品溶液的制备 Preparation of test solution

取本品（鲜品干燥后粉碎）粉末（过三号筛）约lg，精密称定，置具塞锥形瓶中，精密加入甲醇50ml，密塞，称定重量，浸渍20分钟，超声处理（功率250W，频率40kHz）45分钟，放冷，再称定重量，用甲醇补足减失的重量，摇匀，滤过，取续滤液，即得。
Take about 1g of the powder of the product (dried and crushed) (passed through a No. 3 sieve), accurately weigh it, place it in a conical flask with a stopper, accurately add 50ml of methanol, seal tightly, weigh it, soak for 20 minutes, ultrasonicate (power 250W, frequency 40kHz) for 45 minutes, cool, weigh it again, make up for the weight loss with methanol, shake well, and filter. Take the filtrate, and that's it.

测定法 Determination method

分别精密吸取对照品溶液与供试品溶液各20μl，注入液相色谱仪，测定，即得。
Accurately take 20μl of the reference solution and 20μl of the test solution, inject them into the liquid chromatograph, and determine them.

本品按干燥品计算，含毛兰素（C18H22O5）不得少于0.030%。
The product is calculated on the basis of the dried product, and the content of moscatilin (C18H22O5) should not be less than 0.030%.

饮片 Prepared slices

干石斛 Dendrobium

炮制 Processing

除去残根，洗净，切段，干燥。
Remove the residual roots, wash clean, cut into sections, and dry.

性状 Description

本品呈扁圆柱形或圆柱形的段。表面金黄色、绿黄色或棕黄色，有光泽，有深纵沟或纵棱，有的可见棕褐色的节。切面黄白色至黄褐色，有多数散在的筋脉点。气微，味淡或微苦，嚼之有黏性。
This product is in the form of flat cylindrical or cylindrical segments. The surface is golden yellow, greenish yellow or brownish yellow, glossy, with deep longitudinal grooves or longitudinal ribs, some visible brownish brown nodes. The cut surface is yellowish-white to yellowish-brown with most scattered sinewy points. Slight gas, light or slightly bitter taste, chewing viscous.

鲜石斛 Fresh Dendrobium

炮制 Processing

鲜品洗净，切段。
Wash and cut the fresh product.

性状 Description

呈圆柱形或扁圆柱形的段。直径0.4～1.2cm。表面黄绿色，光滑或有纵纹，肉质多汁。气微，味微苦而回甜，嚼之有黏性。
The segments are cylindrical or flat cylindrical. Diameter 0.4 ~ 1.2cm. surface yellowish green, smooth or with longitudinal lines, fleshy and juicy. Gas slightly, taste slightly bitter and sweet, chewing viscous.

鉴别 Identification

（除横切面外）【同药材。
（except cross section）Same as the crude drug.

检查 Examination

同药材。
Same as the crude drug.

霍山石斛 Houshan Dendrobium

性状 Description

同药材。
Same as the crude drug.

鉴别 Identification

同药材。
Same as the crude drug.

检查 Examination

同药材。
Same as the crude drug.

特征图谱 Atlas of features

同药材。
Same as the crude drug.

浸出物 Extractives

同药材。
Same as the crude drug.

含量测定 Assay

同药材。
Same as the crude drug.

性 Property

微寒。
Slightly cold.

味 Flavor

甘。
Sweet.

归经 Meridian tropism

归胃、肾经。
Belongs to the stomach and kidney meridians.

功能 Actions

益胃生津，滋阴清热。
Tonify the stomach, generate body fluids, nourish yin, and clear heat.

主治 Indications

用于热病津伤，口干烦渴，胃阴不足，食少干呕，病后虚热不退，阴虚火旺，骨蒸劳热，目暗不明，筋骨痿软。
Used for heat disease with fluid injury, dry mouth and thirst, insufficient stomach yin, poor appetite and dry vomiting, lingering fever after illness, excessive fire due to yin deficiency, steaming bone and consumptive fever, dim vision, and weakness of tendons and bones.

用量 Dosage

6～12g。鲜品15～30g。
6-12g. Fresh product 15-30g.

用法 Administration

无。
None.

贮藏 Storage

干品置通风干燥处，防潮；鲜品置阴凉潮湿处，防冻。
Preserve dried products in a well-ventilated and dry place, and protect them from moisture; preserve fresh products in a cool and humid place, and protect them from freezing.

铁皮石斛

文本参考：《中国药典（2015年版）》
Text reference: Chinese Pharmacopoeia (2015 Edition)

概述 Overview

本品为兰科植物铁皮石斛Dendrobium officinale Kimura et Migo的干燥茎。11月至翌年3月采收，除去杂质，剪去部分须根，边加热边扭成螺旋形或弹簧状，烘干；或切成段，干燥或低温烘干，前者习称“铁皮枫斗”（耳环石斛）；后者习称“铁皮石斛”。
Dendrobium Stem is the dried stem of Dendrobium officinale Kimura et Migo (Fam. Orchidaceae). The drug is collected from November to the following March, removed from foreign matter, trimmed off some fibrous roots, twisted to a spiral or spring form while heating, and baked to dryness, known as “Tiepifengdou" (Erhuanshihu, earring-like Dendrobium); or cut into sections, then dried or baked to dryness at a low temperature, known as “Tiepishihu".

性状 Description

铁皮枫斗 本品呈螺旋形或弹簧状，通常为2～6个旋纹，茎拉直后长3.5～8cm，直径0.2～0.4cm。表面黄绿色或略带金黄色，有细纵皱纹，节明显，节上有时可见残留的灰白色叶鞘；一端可见茎基部留下的短须根。质坚实，易折断，断面平坦，灰白色至灰绿色，略角质状。气微，味淡，嚼之有黏性。
Tiepifengdou Spiral or spring-like, usually with 2-6 spires, 3.5-8 cm long when stem pulled straight, 0.2-0.4 cm in diameter. Externally yellowish-green or slightly golden, finely wrinkled longitudinally, nodes distinct, sometimes bearing greyish-white leaf sheaths; at one end some short fibrous roots remained at the base of stem. Texture compact, easily broken, fracture even, greyish-white to greyish-green, slightly horny. Odour, slight; taste, weak and viscous on chewing.

铁皮石斛 本品呈圆柱形的段，长短不等。
Tiepishihu Cylindrical sections, various in length.

鉴别 Identification

（1）本品横切面：表皮细胞1列，扁平，外壁及侧壁稍增厚、微木化，外被黄色角质层，有的外层可见无色的薄壁细胞组成的叶鞘层。基本薄壁组织细胞多角形，大小相似，其间散在多数维管束，略排成4～5圈，维管束外韧型，外围排列有厚壁的纤维束，有的外侧小型薄壁细胞中含有硅度块。含草酸钙针晶束的黏液细胞多见于近表皮处。
（2）取本品粉末1g，加三氯甲烷-甲醇（9：1）混合溶液15ml，超声处理20分钟，滤过，滤液作为供试品溶液。另取铁皮石斛对照药材1g，同法制成对照药材溶液，照薄层色谱法（通则0502）试验，吸取上述两种溶液各2～5μl，分别点于同硅胶G薄层板上，以甲苯-甲酸乙酯-甲酸（6：3：1）为展开剂，展开，取出，烘干，喷以10%硫酸乙醇溶液，在95℃加热约3分钟，置紫外光灯（365nm）下检视。供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的荧光斑点。

检查 Examination

水分 Water

不得过12.0%（通则0832第二法）。
Not more than 12.0 per cent <0832, method 2>.

总灰分 Total ash

不得过6.0%（通则2302）。
Not more than 6.0 per cent <2302>.

浸出物 Extractives

照醇溶性浸出物测定法（通则2201）项下的热浸法测定，用乙醇作溶剂，不得少于6.5%。
Carry out the method for determination of ethanol-soluble extractives <2201, the hot extraction method>, using ethanol as the solvent, not less than 6.5 per cent.

含量测定 Assay

多糖 对照品溶液的制备 取无水葡萄糖对照品适量，精密称定，加水制成每1ml含90μg的溶液，即得。标准曲线的制备精密量取对照品溶液0.2ml、0.4ml、0.6ml、0.8ml、1.0ml，分别置10ml具塞试管中，各加水补至1.0ml，精密加入5%苯酚溶液1ml（临用配制），摇匀，再精密加硫酸5ml，摇匀，置沸水浴中加热20分钟，取出，置冰浴中冷却5分钟，以相应试剂为空白，照紫外-可见分光光度法（通则0401），在488nm的波长处测定吸光度，以吸光度为纵坐标，浓度为横坐标，绘制标准曲线。
供试品溶液的制备 取本品粉末（过三号筛）约0.3g，精密称定，加水200ml，加热回流2小时，放冷，转移至250ml量瓶中，用少量水分次洗涤容器，洗液并人同一量瓶中，加水至刻度，摇匀，滤过，精密量取续滤液2ml，置15ml离心管中，精密加入无水乙醇10ml，摇匀，冷藏1小时，取出，离心（转速为每分钟4000转）20分钟，弃去上清液（必要时滤过），沉淀加80%乙醇洗涤2次，每次8ml，离心，弃去上清液，沉淀加热水溶解，转移至25ml量瓶中，放冷，加水至刻度，摇匀，即得。

性味与归经 Property and Flavor

甘，微寒。归胃、肾经。
Mild cold; sweet. Stomach and kidney meridians.

功能与主治 Actions and Indications

益胃生津，滋阴清热。用于热病津伤，口干烦渴，胃阴不足，食少干呕，病后虚热不退，阴虚火旺，骨蒸劳热，目暗不明，筋骨痿软。
To boost the stomach, engender fluids, nourish yin, and clear heat. Used for heat disease consuming fluid, dry mouth, vexation and thirst, stomach yin deficiency, reduced food intake, retching, deficiency heat after recovering from an illness, yin deficiency with effulgent fire, steaming bone and consumptive fever, dim and blurred vision, limp wilting sinew and bone.

用法与用量 Administration and Dosage

6～12g。
6-12 g.

贮藏 Storage

置通风干燥处，防潮。
Preserve in a ventilated and dry place, and protect from moisture.