• 154.50 KB
  • 20页

荧光素钠注射液的制备及质量检查 毕业论文

  • 20页
  • 当前文档由用户上传发布,收益归属用户
  1. 1、本文档共5页,可阅读全部内容。
  2. 2、本文档内容版权归属内容提供方,所产生的收益全部归内容提供方所有。如果您对本文有版权争议,可选择认领,认领后既往收益都归您。
  3. 3、本文档由用户上传,本站不保证质量和数量令人满意,可能有诸多瑕疵,付费之前,请仔细先通过免费阅读内容等途径辨别内容交易风险。如存在严重挂羊头卖狗肉之情形,可联系本站下载客服投诉处理。
  4. 文档侵权举报电话:19940600175。
'吉林省高等教育自学考试吉林大学生物制药专业本科毕业设计设计题目:荧光素钠注射液的制备及质量检查设计作者:准考证号:通信地址:联系电话:吉林大学生命科学学院2012年3月24日 摘要本文研究了荧光素钠注射液的制备及质量检查。首先对荧光素钠进行提纯和检查,并对荧光素钠进行酸碱度,溶液的澄清度和颜色,氯化物,硫酸盐,锌盐,干燥失重等检查。采用重量测定法测定含量。对制备工艺中的活性炭用量筛选、pH值的选择、灭菌温度、处方制备工艺等进行考察。后对荧光素钠注射液进行光照(4000lx),高温(60℃),低温(4℃)等影响因素实验并检测含量变化。最后对荧光素钠注射液进行性状检查和鉴别,采用中国药典规定的重量法测定含量。结果表明:本制备工艺科学合理,产品质量好,注射液的pH值、可见异物均符合要求。采用重量法测定荧光素钠的含量,其重复性试验、重现性试验、回收率试验均符合测试要求,说明用本法测定荧光素钠的含量准确度好。测定样品荧光素钠的含量为标示含量的99.5%,符合为标示含量的90.0%~110.0%的范围,符合要求。关键词:荧光素钠;荧光素钠注射液;制备工艺;含量检查I-21- 目录一、前言·····································································································1㈠试剂及仪器·······················································································21.原料与试剂··························································································22.仪器···································································································2㈡实验方法与步骤···················································································21.荧光素钠提纯························································································22.荧光素钠原料检查·················································································3a.性状·····································································································3b.鉴别·····································································································4c.检查·····································································································5-21- d.含量测定······························································································73.荧光素钠注射液的制备···········································································8a.荧光素钠注射液的配制···········································································8b.制备工艺流程图···················································································8c.处方依据······························································································8d.工艺筛选······························································································9e.影响因素试验······················································································10f.处方中各成分的作用·············································································114.制剂检测·····························································································11a.性状·································································································11-21- b.鉴别·································································································11c.检查·································································································12d.含量测定···························································································13㈢结果与讨论·························································································141.荧光素钠静脉注射液的PH值··································································142.注射液中微粒及细菌·············································································15㈢参考文献·····························································································16㈣致谢································································································17前言-21- 荧光素钠注射液用于供诊断眼角膜损伤、溃疡和异物,眼底血管造影和循环时间测定。也用于术中显示胆囊和胆管,以及结核性脑膜炎的辅助诊断等。主要用于:1.循环时间测定(前臂静脉注射,成人常用量5ml(10%);小儿常用量,按体重0.05ml/kg(10%),全量在1秒内快速推入。);2.眼底血管造影(静脉注射,成人常用量5ml(10%),或按体重15~30mg/kg计算,全量在4秒左右推注完毕,注射后8秒开始在蓝色光波激发下用荧光眼底照相机连续摄影,开始每秒1张,连续10秒,以后在30分钟内适当间隔摄片,也可用眼底镜直接观察。);3.术中显示胆囊和胆管(手术前4小时静脉注射5ml(10%)。);4.脑脊液渗透率试验(诊断结核性脑膜炎。肌内注射,成人推荐用量5~10ml(10%);儿童推荐用量按体重0.3ml/kg(10%),注射后2小时腰椎穿刺抽取脑脊液,与浓度为0.00001%、0.00002%、0.00003%、0.00005%、0.00006%、0.0001%、0.0002%和0.0005%的标准系列比色管比色。);另外还用于眼科检查角膜损伤、溃疡或异物(作2%滴眼液以微细滴管滴眼)。其中眼角膜损伤和血管造影诊断及血液循环时间的测定[1,2],显影清晰度和持续时间是造影的重要质量指标。药理学研究显示:本品为诊断用药,是一种染料,对正常角膜等上皮不能染色,但能对损伤的角膜上皮染成绿色,从而可显示出角膜损伤、溃疡等病变。本品流经小血管时,能在紫外线或蓝色光激发下透过菲薄的血管壁和粘膜呈现绿色荧光,从而显示小血管行经和形态等,据此可供眼底血管造影和循环时间测定。荧光素钠静脉注射液是由间-苯二酚和邻-苯二甲酸酐合成为荧光素粗制品,经提纯成为精品后用氢氧化钠、注射水配制而成的。本品过敏反应包括:荨麻疹、呼吸困难、哮喘发作、呼吸停止、血压下降、休克、心脏停搏、心肌梗塞、肺水肿和脑梗塞等。常见反应有恶心、呕吐、眩晕,多在注射后30秒钟内发生。反应发生率和严重程度与注射剂浓度和注入量有关。一次静脉注入量>5ml(5%),常可出现恶心和呕吐症状[3]。本品血管造影时总的反应发生率约0.6%,严重反应约0.4%。静脉注射后皮肤和尿液暂时染色,视物有黄色或粉红色感觉。本实验研究了荧光素钠注射液的制备工艺及含量的检测,检测结果符合要求,说明方法可行。荧光素钠注射液的制备及质量检查㈠试剂及仪器1.原料与试剂氯化铵(分析纯国药集团化学试剂有限公司);氢氧化钠(分析纯国药集团化学试剂有限公司);活性炭(药用国药集团化学试剂有限公司);盐酸(分析纯-21- 沈阳市新化试剂厂);甲醇(分析纯沈阳市新化试剂厂);乙醇(分析纯沈阳市新化试剂厂);氯仿(分析纯沈阳市新化试剂厂);重铬酸钾(分析纯沈阳新化试剂厂);浓硫酸(分析纯沈阳市新化试剂厂);硝酸(分析纯沈阳市新化试剂厂);磷酸(分析纯沈阳市新化试剂厂);荧光素钠注射液(自制,规格0.6g:3mL);磷酸二氢钠(分析纯国药集团化学试剂有限公司);氯化钠(分析纯国药集团化学试剂有限公司);丁醇(分析纯沈阳市新化试剂厂);异丁醇(分析纯沈阳市新化试剂厂);试验样品(批号:yp01,yp02,yp03):均为自制。对照品:系由原料药重结晶制得,纯度为99.6%,含量为99.7%(重量法)。市售样品:荧光素钠注射液20瓶(广州明兴制药有限公司生产,批号:031102,0.6g/3ml),合并后蒸出大部分水,剩余物冷却后抽滤,用少量水洗涤,120℃真空干燥,得市售样品。2.仪器UV9200型紫外一可见分光光度计(北京瑞利分析仪器公司);JA-1003型电子分析天平(北京赛多利斯仪器系统有限公司);电热鼓风干燥箱(上海博迅实业有限医疗设备厂);雷磁pHS-3C型精密pH计SC-4000A型澄明度检测仪(上海黄海药检仪器有限公司)㈡实验方法与步骤1.荧光素钠提纯取荧光素粗制品,用适量的氢氧化钠加热溶解,然后加人l%活性炭(按溶液体积计算),搅拌15分钟,过滤。滤液加酸得荧光素沉淀,静置3小时,抽滤。沉淀用注射用水洗涤。然后把沉淀再用碱溶解、过滤、加酸沉淀、洗涤,如此反复3次,得到荧光素钠精品,最后把精品烘干、备用。2.荧光素钠原料检查a、性状(1)外观、臭、味:本品为橙红色粉末;无臭,几乎无味。(2)引湿性:取适量,敞口置于25℃±1℃恒温干燥器(氯化铵饱和溶液)内,放置24小时,结果见表2-1。-21- 表2-1荧光素钠引湿性试验结果批号yp01yp02yp03市售样品吸湿增重%13.813.012.814.2由试验结果可知,本品具有引湿性。(3)溶解度:照中国药典2005年版二部凡例溶解度项下的有关规定试验,荧光素钠对照品在常见溶剂中溶解度[4]见表2-2。表2-2溶解度试验结果溶剂对照品量(mg)对照品量/溶剂量(g/ml)溶解情况水1001:9易溶甲醇1001:9易溶乙醇1001:90略溶氯仿1001:10000几乎不溶0.1mol/LNaOH1001:0.9极易溶解由试验结果可见,本品在水、甲醇中易溶,在乙醇略溶,在氯仿中几乎不溶,在0.1mol/LNaOH中极易溶解。经过三批样品试验,溶解度均符合规定。(4)吸收系数:①仪器的校正和检定:对所用UV9200型紫外一可见分光光度计在测定前进行全面检定,吸收度的准确度按中国药典2005年版二部附录IVA项下,用重铬酸钾的硫酸溶液检定,结果见表2-3。仪器实测的吸收系数与规定值的相对偏差小于±1%,符合规定。表2-3仪器吸收度校正结果波长/nm235(最小)257(最大)313(最小)350(最大)规定E1%1cm124.5144.048.62106.6吸收度0.7630.8810.2970.651E1%1cm124.6143.848.49106.3-21- 相对偏差(%)0.080.210.270.28②紫外图谱的测定:取荧光素钠对照品适量,用甲醇为溶剂,制成每1ml含8μg的溶液,照分光光度法(中国药典2005年版二部附录ⅣA),在200~400nm波长范围内扫描,,结果表明本品在321nm±1nm,277.5nm±1nm,234nm±1nm处有吸收峰。③方法与结果:供试溶液的制备:精密称取对照品25mg,置100ml量瓶中,加甲醇溶解并稀释至刻度,摇匀,作为贮备液。精密量取贮备液2ml(溶液1,A=0.3~0.4)和4ml(溶液2,A=0.6~0.8),置100ml量瓶中,用甲醇稀释至刻度,摇匀,即得。吸收系数的测定:温度为18℃~20℃,用1cm石英吸收池,在234nm波长处分别测定吸收度,计算吸收系数,吸收系数测定结果见表2-4,2-5。表2-4吸收系数的测定数据吸收度(A)试样I试样II溶液15.07μg/ml溶液210.04μg/ml溶液34.99μg/ml溶液49.98μg/ml0.3610.7240.3570.716表2-5吸收系数的测定结果吸收系数(E1%1cm)试样I试样II溶液1溶液2溶液3溶液4712.0714.0715.4717.4X±SD714.7±16.27测得吸收系数(E1%1cm)为714.7(RSD=2.08%),经对三批样品测定,吸收系数分别为:yp01,714.2;yp02,715.1;yp03,714.5。 b、鉴别化学鉴别法:(1)-21- 本品的水溶液显强烈的荧光,用大量的水稀释后仍极明显;但加酸使成酸性后,荧光即消失;再加碱使成碱性,荧光又显出。结果:三批样品均成正反应。(2)本品炽灼灰化后显钠盐的鉴别反应。结果:三批样品均成正反应。 c、检查(1)酸碱度:①仪器与试剂:雷磁PHS-3C型精密PH计②测定方法:取三批样品与对照品各约1.0g,加水溶解并稀释至50ml,依法测定(中国药典2005年版二部附录VIH)pH值应为7.0~9.0。③结果:结果见表2-6。表2-6荧光素钠pH值测定结果批号yp01yp02yp03市售样品PH值7.917.958.038.06(2)溶液的澄清度和颜色:取三批供试品与对照品各1.0g,加水50ml使溶解,观察溶液的颜色与澄清度。结果见表2-7表2-7荧光素钠澄清度和颜色检查结果批号yp01yp02yp03市售样品澄清度澄清澄清澄清澄清颜色橙黄色并有黄绿色荧光橙黄色并有黄绿色荧光橙黄色并有黄绿色荧光橙黄色并有黄绿色荧光结论:本品三批原料药与对照品的水溶液均澄清,颜色为橙黄色并有黄绿色荧光。(3)氯化物:①供试溶液:取本品0.10g,加水50ml使溶解,分取溶液10ml,加水25ml和稀硝酸10ml,过滤;置50ml纳氏比色管中,加水使成约40ml,摇匀,即得供试溶液。②测试操作方法:取上述供试溶液,按(中国药典2005年版二部附录ⅧA)氯化物检查法操作,和标准氯化钠溶液7.0ml制成的对照溶液进行比较,不得更浓(0.35%)。③结果:结果见表2-8。-21- 表2-8荧光素钠原料药氯化物检查批号yp01yp02yp03市售样品氯化物<0.35%<0.35%<0.35%<0.35%(4)硫酸盐:①供试溶液:取上述氯化物项下剩余的溶液25ml,置50ml纳氏比色管中,加水使成约40ml,过滤,加稀盐酸2ml,摇匀,即得供试品溶液。②测试操作方法:取上述供试溶液,按(中国药典2005年版二部附录ⅧB)硫酸盐检查法操作,和标准硫酸钾溶液2.5ml制成的对照溶液进行比较,不得更浓(0.50%)。③结果:结果见表2-9。表2-9荧光素钠原料药硫酸盐检查结果批号结果yp01<0.50%yp02<0.50%yp03<0.50%市售样品<0.50%(5)锌盐:①测试操作方法:取本品约0.10g,加氯化钠饱和水溶液10ml溶解后,加稀盐酸2ml,摇匀,滤过,滤液中加亚铁氰化钾试液1ml,不得发生浑浊。②结果:结果见表2-10。表2-10荧光素钠原料药锌盐检查批号yp01yp02yp03市售样品锌盐未见浑浊未见浑浊未见浑浊未见浑浊(6)干燥失重:①仪器:电热鼓风干燥箱②测定方法:取本品,照干燥失重测定法(中国药典2005年版二部附录ⅧL)操作,在105℃干燥至恒重,减失重量不得过7.0%。-21- ③结果:结果见表2-11。表2-11干燥失重检查结果批号yp01yp02yp03市售样品干燥失重1.29%1.37%1.34%1.18%结论:三批样品及对照品干燥失重均小于7.0%。d、含量测定荧光素钠原料的含量测定照荧光素钠含量测定法(中国药典2005年版二部附录P448),采用重量法。(1)测定法:取本品约0.5g,精密称定,加水20ml溶解后,加稀盐酸5ml使荧光素析出,用丁醇-氯仿(1:1)提取4次,每次20ml,合并提取液,用水10ml洗涤,洗液再用异丁醇-氯仿(1:1)5ml振摇提取,合并提取液,置105℃恒重的容器中,在水浴上通风蒸发至干,残渣用乙醇10ml溶解后,再置水浴上蒸干,并在105℃干燥至恒重,精密称定,残渣重量与1.132相乘,即得供试量中含有C20H10Na2O5的重量。(2)重复性试验:再取本品,精密称定,照含量测定方法测定含量。共测6份。结果见表2-12。表2-12重复性试验结果序号123456含量(%)99.199.599.899.299.199.0含量平均值(%)99.3RSD%0.31(3)样品测定:照测定法,对三批样品进行测定,测定结果见表2-13。表2-13三批样品含量测定批号yp01yp02yp03含量(%)99.499.699.53、荧光素钠注射液的制备-21- a、荧光素钠注射液的配制(1)称取处方量的荧光素钠,加入注射用水近全量,搅拌,加热使完全溶解;(2)滴加0.1mol/L氢氧化钠溶液适量,调节pH至8.2~8.4,加入注射用水至全量;(3)加入0.1%的药用活性炭,80℃保温搅拌20min,趁热脱炭;(4)测定半成品含量,符合要求后药液经0.45μm的微孔滤膜过滤,灌装至5ml安瓿中;(5)密封,于100℃流通蒸气灭菌40min;(6)灯检,印字。b、制备工艺流程图[6]溶液1荧光素钠加注射用水0.1mol/LNaOH调节pH至8.2~8.4半成品溶液2加0.1%药用活性炭测含量,经0.45μm保温20分钟,搅拌,微孔滤膜滤过趁热脱炭溶液3成品溶液4灌装,密封灯检,印字c、处方依据1)规格依据:根据荧光素钠注射液(中国药典2005年版二部)规格项下规定,确定其规格为3ml:0.6g。2)pH:根据荧光素钠注射液质量标准(中国药典2005年版二部)pH值的要求(8.0~9.8)以及一般注射剂对pH值的要求,确定本品在溶液配制时的pH值控制在8.2~8.4。d、工艺筛选(1)活性炭用量筛选配制注射液过程中加入活性炭可以吸附药液的杂质和热原。考察加入0.05%,0.1%和0.15%的针用活性炭后,溶液中荧光素钠含量的变化情况。-21- 表3-1活性炭用量的选择活性炭用量(%)加入活性炭前含量(%)加入活性炭后含量(%)0.0599.899.30.199.898.90.1599.898.7从试验中可见活性炭吸附有效成分含量,从试验结果并参照注射液的制备工艺,确定加活性炭的量为0.1%,为保证终产品热原检查合格,用0.1%的活性炭对制剂中间溶液进行处理,可以达到去除热原的目的,并在制备过程中应测定中间体含量并应调到规定的浓度即可。(2)pH值的选择分别配制pH5、pH6、pH7、pH8、pH9的荧光素钠注射液,分别在4℃时放置荧光素钠注射液10天,取样观察其溶液的颜色及澄明度。取样观察pH5、6的荧光素钠注射液有沉淀析出,pH7、pH8、pH9的荧光素钠注射液中溶液澄明。表3-2pH值选择考察结果pH56789澄明度有沉淀析出有沉淀析出澄明澄明澄明颜色橙黄色橙黄色棕色棕色棕色从以上结果可以看出荧光素钠注射液在pH7~9之间较稳定。(3)灭菌温度取按上述制备工艺制备的荧光素钠注射液,分别于灭菌温度115℃,灭菌时间30min;灭菌温度121℃,灭菌时间20min;灭菌温度126℃,灭菌时间15min后,放置,检查,结果表明,选择115℃灭菌30min比较适合。表3-3灭菌条件考察情况检查条件颜色灭菌前pH灭菌后pH澄明度灭菌温度115℃灭菌时间30min棕色8.268.24澄明灭菌温度121℃灭菌时间20min棕色8.268.29澄明-21- 灭菌温度126℃灭菌时间15min棕色8.268.25澄明(4)处方制备工艺称取处方量的荧光素钠,加入注射用水至全量,搅拌,加热使完全溶解;加0.1mol/L氢氧化钠溶液适量,调节溶液至pH8.2~8.4,加入注射用水至全量;加入0.1%的药用活性炭,保温搅拌20min,趁热脱炭;测定半成品含量,经检验合格后,药液经0.45μm的微孔滤膜过滤,灌装至5ml安瓿中,于115℃流通蒸气灭菌30min,灯检,印字,即得。e、影响因素试验制备的样品进行影响因素试验,以考察稳定性。(1)光照试验(4000lx)方法:室温条件下,取样品,在照度为4000lx的光橱中放置10天,于第5天和第10天取样,对外观色泽、澄明度、pH值、含量等项目进行考察,并将结果与0天资料比较。结果:见表3-4。(2)高温试验(60℃)方法:取样品,置60℃恒温箱中放置10天,于第5天和第10天取样,对外观色泽、澄明度、pH值、含量等项目进行考察,并将结果与0天资料比较。结果:见表3-4。(3)低温试验(4℃)方法:取样品,置4℃冰箱中放置2天,然后在40℃加速条件下考察2天,循环3次,取样检查对外观色泽、澄明度、pH值、、含量等项目进行考察,并将结果与0天资料比较。结果:见表3-4。表3-4影响因素试验结果考察项目-21- 试验条件观察时间外观色泽pH澄明度含量(标示量%)0天棕色澄明液体8.26符合规定99.5光照(4000lx)5天棕色澄明液体8.28符合规定99.310天棕色澄明液体8.23符合规定99.4高温(60℃)5天棕色澄明液体8.25符合规定99.410天棕色澄明液体8.24符合规定99.2低温(4℃)12天棕色澄明液体8.22符合规定99.0结果表明:在光照(4000lx)、60℃高温下放置10天及在4℃低温条件下进行试验,外观色泽、澄明度、pH值、和含量均未见明显变化。f、处方中各成分的作用1)荧光素钠:主药2)注射用水:溶媒3)氢氧化钠:调pH4.制剂检查a、性状方法:取样品,目视观察。结果:样品为棕色澄明液体。表4-1性状检查结果批号zj01市售样品性状棕色澄明液体棕色澄明液体b、鉴别(1)色谱鉴别原理:样品中荧光素钠的HPLC主峰保留时间应与对照品主峰保留时间相同[7]。方法:取样品按有关物质测定项下方法记录色谱图。-21- 结果:见表4-2。样品主峰保留时间均与对照品主峰保留时间相同。表4-2色谱鉴别结果批号zj01市售样品tR(min)7.4437.460(2)化学鉴别法:本品的水溶液显强烈的荧光,用大量的水稀释后仍极明显;但加酸使成酸性后,荧光即消失;再加碱使成碱性,荧光又显出。结果:空白辅料对其无干扰,样品成正反应。c、检查(1)pH值仪器:雷磁pHS-3C精密酸度计.方法:取样品3瓶,依法测定(中国药典2005年版二部附录ⅣH)。结果:见表4-3。样品的pH值在8.0~9.8。表4-3pH值检查结果批号zj01市售样品pH值8.348.29(2)可见异物取本品,按照中国药典2005年版二部附录制剂通则中注射剂的要求,进行澄明度检查。方法:取样品2瓶,采用YB-2型澄明度检测仪(天津大学精密仪器厂制造)进行检查。结果:样品符合规定。(3)装量取样品,依法检查(按中国药典2005年版二部附录ⅩF)。结果:见表4-4。装量均符合规定。表4-4装量检查结果-21- 批号装量(ml)装量(ml)装量(ml)平均装量(ml)结果zj013.163.203.183.18均符合规定d、含量测定(1)荧光素钠的含量测定[8]方法:采用重量法进行测定。参照中国药典2005年版二部P449荧光素钠含量测定进行测定。测定法:取本品(相当于荧光素钠约0.5g),加水稀释至20ml摇匀,加稀盐酸5ml使荧光素析出,用丁醇-氯仿(1:1)提取4次,每次20ml,合并提取液,用水10ml洗涤,洗液再用异丁醇-氯仿(1:1)5ml振摇提取,合并提取液,置105℃恒重的容器中,在水浴上通风蒸发至干,残渣用乙醇10ml溶解后,再置水浴上蒸干,并在105℃干燥至恒重,精密称定,残渣重量与1.132相乘,即得供试量中含有C20H10Na2O5的重量。(2)方法学研究[9,10]①重复性试验同一份供试品溶液测定6次,结果见表4-5。表4-5重复性试验结果序号123456RSD(%)含量(%)99.599.199.699.299.399.50.24结论:该法重复性较好,符合测试要求。②重现性试验由不同试验人员进行重现性试验。结果见表4-6。表4-6重现性试验结果序号123RSD%含量(%)98.999.699.30.35③回收率试验取荧光素钠原料药(yp01,含量99.4%),照处方中主药的比例,按10瓶的配制量配制,取80%、100%和120%量的主药荧光素钠-21- 各三份,精密称定,分别按处方量的100%加入注射用水。按上述含量测定方法测定荧光素钠的含量,计算回收率。结果见表4-7。表4-7回收率试验结果加入量(g)测得量(g)回收率(%)平均回收率(%)RSD(%)4.81424.189399.199.40.314.98134.343499.35.08984.455999.76.01935.274999.899.50.316.02125.244999.26.01275.258699.67.18136.274399.599.40.167.19246.265199.27.28926.362299.4结论:用本法测定本品中荧光素钠的含量,回收率在98.0%~102.0%,准确度好。(3)样品含量测定取样品,照含量测定方法测定含量,结果见表4-8。.表4-8含量测定结果样品批号标示量(%)zj0199.5样品荧光素钠的含量为标示含量的90.0%~110.0%。㈢、结果与讨论1.荧光素钠注射液的pH值美国药典规定pH为8.0—9.0[8];当溶液pH值大于8.5时,易产生静脉炎[6]。根据上述文章及本研究的资料,为了既能获得荧光度强又符合注射要求的注射液,故本制剂的pH值控制在8.2—8.4范围内。2.注射液中微粒及细菌-21- 注射液中的微粒是造成患者副作用的主要因素之一,已受到国内外有关专家的重视,为消除微粒及细菌的危害,采用孔径为0.45微米的滤膜过滤并经加热灭菌,即可达到要求。参考文献[1]MorinCJ,MofaddelNL,DesbeneAM.Utilizationoffluoresceinsodiumsaltforthe-21- indirectfluorimetricdetectioninmicellarelectrokineticchromatography[J].JournalofChromatographyA,2007,(872):247-258.[2]TsutomuHara.Efficacyandsafetyoffluoresceinangiographywithorallyadministeredsodiumfluorescein[J].AmericanJournalofphthalmology,1998,126(4):560-564.[3]宋志燕,蒋菊蓉,李钰珍.护理程序在眼底荧光血管造影中的应用[J].护理研究,2005,19(5):890-891.[4]中国药典2005版二部288.[5]W.R.OrndorefetalJ.chemicalReview.1997.49.127.[6]刘玉兰,姚丽,张吉平,等.荧光素钠注射液的制备及质量控制[J].山东医药工业杂志,1997,16(6):3-5.[7]代红,田洪昭,余立.反相高效液相色谱法测定荧光素钠注射液[J].药物分析杂志,2000,20(4):267-268.[8]牛桂田,杨振宇,丁长生.重量法测定荧光素钠及制剂的含量[J].药物分析杂志,2006,14(2):55-56.[9]MckennaJF,SowaFJ.Organicreactionswithboronfluoride[J].JAmChemSoc,2007,60:124-125.[10]WoodroofeCC,LimMH,BuW,etal.Synthesisofi-somericallypurecarboxylate-andsulfonate-substitutedxanthenefluorophores[J].Tetrahedron,2005,61:3097-3105.致谢-21- 本论文是在导师的悉心指导下完成的。导师从开始的课题设计、实验操作到最后的论文的撰写、修改都倾注了大量的心血。导师的谆谆教诲和无微不至的关怀为我的成长提供了莫大的帮助,并将始终激励着我不断地努力进取。导师严谨求实的科研作风、诲人不倦的教学态度以及对我的关心和指导令我铭记在心,终身难忘。导师严谨的治学态度、渊博的学识、高尚的品德和豁达的胸怀永远是我学习、工作的榜样。在此向老师表示衷心的感谢。最后,特别感谢百忙中对我的论文进行细心审阅的各位老师,并感谢答辩委员会各位老师提出宝贵意见。-21-'