药物分析光谱实验讲义 光谱实验(版本1目前用的).doc

药物光谱分析实验提示：本次实验所用仪器均为贵重精密仪器，仪器操作前务必仔细阅读仪器旁的操作规程！实验一 葡萄糖比旋度的测定1、取葡萄糖约10g，精密称定，置100ml量瓶中，加水适量与氨试液0.2ml，溶解后，用水稀释至刻度，摇匀，放置10分钟，在25时，依法测定，比旋度应为+52.5至+53.0。2、注意事项：测定旋光度时，应同上法制备不含葡萄糖的空白溶液，将测定管用供试液冲洗数次，缓缓注入供试液（注意避免产生气泡），置于旋光计内读出旋光度，测定空白管和供试品管时旋光管方向应一致。读数时注意，仪器会自动给出三个读数，并计算其平均值及SD，CV等，仪器读数过程中千万不可打开样品仓盖，等三个数据都给出后才可开盖取出旋光管，否则容易损坏仪器。比旋度的计算： ：比旋度；l：测定管长度(dm)；c：每100ml溶液中含有被测物质的重量（g）实验二 磺胺甲噁唑的紫外分光光度法鉴别1、称取磺胺甲噁唑对照品约50mg,，置100ml的量瓶中，加乙醇溶解并稀释至刻度，摇匀。得对照品溶液。2、碱性供试品溶液的制备：精密量取上述对照品溶液2.0ml置100ml量瓶中，加0.1 mol/L氢氧化钠溶液稀释至刻度，摇匀，得到碱性供试品溶液，同法制备空白溶液，扫描紫外吸收曲线。3、酸性供试品溶液的制备：精密量取上述对照品溶液2.0ml置100ml量瓶中，加0.01 mol/L盐酸溶液稀释至刻度，摇匀，得到酸性供试品溶液，同法制备空白溶液，扫描紫外吸收曲线。4、注意事项：紫外扫描时先在参比池和样品池中均放入空白溶液，进行基线校正（Basecorrect），校正完毕后参比池不动，把样品池（靠近操作者的比色皿）换成供试品溶液进行扫描。实验三 维生素B1的荧光扫描1、氧化试剂的制备：取新鲜配制的1.0铁氰化钾溶液4.0ml，加3.5 mol/L氢氧化钠溶液制成100ml，于4h内使用。2、对照品溶液的制备：精密吸取对照品储备液1.0ml，置10ml量瓶中，用0.2mol/L盐酸溶液稀释至刻度，摇匀，得2 g/ml的溶液，再精密吸取2 g/ml的溶液1.0ml，置10ml量瓶中，用0.2mol/L盐酸溶液稀释至刻度，摇匀，得0.2 g/ml的溶液，即得。3、测定法：取50ml量瓶，精密加入对照品溶液5ml，迅速加入（12s内）氧化试剂3.0ml，在30s内再加入正丁醇20.0ml，密塞，剧烈振摇90s；再加入无水乙醇2ml，旋摇数秒，待分层后，取上层澄清的正丁醇液约10ml，置荧光计测定池内，进行荧光扫描，确定最大激发波长（excitation wavelength）和发射波长（emission wavelength）。Exp.1 The Identification of IR Spectro-photometric Method for Propranolol Hydrochloride1. Preparation of sampleBlank tablet: Weight about 200 mg dry KBr powder, grind and mix well, then transfer to the mould, make the tablet.Sample tablet: Weight about 1 mg Propranolol Hydrochloride and 200 mg dry KBr powder, grind and mix well, then transfer to the mould, make the tablet.2. DeterminationPlace the sample tablet in the sample light route of the instrument, deduct the background of the blank tablet, record the IR spectro-photometric figure.The infrared absorption sectrum is concordant with the reference spectrum. Exp. 2 The Determination of Specific Optical Rotation of GlucoseDissolve, about 10 g, weighed accurately, with a quantity of water and 0.2 ml of ammonia TS in a 100 ml volumetric flask and dilute with water to volume. Mix well and allow to stand for 10 minutes. The specific optical rotation of the resulting solution is +52.5 +53.0at 25.Exp.3 The Identification of UV Spectrophotometric Method for Sulfamethoxazole1. Preparation of standard solution: Weigh about 50 mg sulfamethoxazole CRS and transfer to 100 ml volumetric flask, dissolve in ethanol and dilute to volume, mix well. 2. Preparation of alkaline sample solution: Transfer 2 ml of that solution to 100 ml volumetric flask, dilute with 0.1 mol/L sodium hydroxide solution to volume and mix well. Perform blank solution using the same method. Then scan the UV absorption curve. 3. Preparation of acidic sample solution: Transfer 2 ml of that solution to 100 ml volumetric flask, dilute with 0.01 mol/L hydrochloric acid solution to volume and mix well. Perform blank solution using the same method. Then scan the UV absorption curve. 4. Attention: Place the blank solution in the reference cell and sample cell, respectively. And conduct baseline correction. Then change the solution of sample cell (near the operators cuvette) into the sample solution. Exp.4 The Scan of FR Spectro-photometric for Vitamin B11. Preparation of Oxidation Reagent: Prepare 1.0 % red potassium prussiate 4.0 ml freshly, add 3.5 mol/L sodium hydroxide to 100 ml, used in 4 hours.2. Preparation of standard solution: Weigh accurately 20 mg vatamin B1 CRS, dissovle in 300 ml diluent ethanol (15), adjust pH to 4.0 with 3mol/L hydrochloric acid, and then dilute in diluent ethanol to 1000 ml as the store solution that can store for 1 month at low temprature and free from the light. Transfer 1.0 ml store solution to 10 ml volumetric flask, dissolve in 0.2mol/L hydrochloric acid and dilute to volume, mix well and get the 2 g/ml solution. Then transfer 2 g/ml solution 1.0 ml to 10 ml volumetric flask, dissolve in 0.2mol/L hydrochloric acid and dilute to volume, mix well and get the 0.2 g/ml solution.3. Transter 5.0 ml standard solution to 50 ml volumetric flask, add 3.0ml oxidation reagent rapidly (within 12s ), add 20.0 ml isobutyl alcohol in 3