摘要
做为畜禽专用抗生素,替米考星具有极好的抗菌活性及药代动力学特征,其治疗畜禽感染疗效确切,效果显著,使用安全,但是由于其味道极苦,动物服用相当困难,而且替米考星禁止静注,因而对替米考星的使用带来一定影响。
本课题首次对替米考星海藻酸钙凝胶微丸的制备工艺进行了考察。实验中以载药量和包封率为指标,通过单因素试验,对海藻酸钠浓度、氯化钙浓度、钙化时间、滴制参数、海藻酸钠与替米考星的配比等影响因素进行研究。通过海藻酸钠与替米考星的配比、氯化钙浓度、海藻酸钠浓度、钙化时间进行四因素三水平的正交试验。实验发现海藻酸钠浓度2%、氯化钙浓度2%、钙化时间1h左右、滴制参数(针头离液面的距离8cm左右、滴制的速度30d/min、针头型号为12号)、海藻酸钠与替米考星的比例1:5为最佳的制备工艺。在选定的制备工艺条件下可制备得到外观较好,载药量和包封率也较高的替米考星海藻酸钙凝胶微丸,而且降低了药物的苦味。利用高效液相色谱法建立替米考星海藻酸钙凝胶微丸的含量检测标准。采用反相高效液相法,用水-乙腈-四氢呋喃-二丁胺磷酸盐缓冲液(805:115:55:25)为流动相,柱温:30℃,检测波长:280nm,进样量:10μl。用外标峰面积法定量,以顺式和反式峰面积之和(Y)为纵坐标,替米考星的浓度(X)为横坐标作图,得一直线,求得回归方程,计算相关系数。得标准曲线方程为:Y=565252X+53867,R~2=0.9993。X在0.25—0.85mg/ml范围内呈良好线性关系。测得替米考星的含量为76.13%。通过含量测定、性状、鉴别、溶散时限的研究,为考察制剂的稳定性提供准确精密的方法,为制定制剂质量标准提供依据。本论文并对凝胶微丸的稳定性进行了加速试验和长期试验。通过高温试验、高湿试验、强光试验等初步考察了影响稳定性的几个主要因素。研究结果表明:微丸对光照比较敏感,故应避光保存;微丸在常温下具有较好的稳定性,但在高温60℃条件下加热24小时,微丸的色泽、含量均有很大改变,提示微丸应室温储存。
本实验研究的替米考星海藻酸钙凝胶微丸的制备工艺成熟、可靠,具有较高的包封率和载药量,能有效地掩蔽替米考星的苦味;所建立的HPLC检测替米考星海在藻酸钙凝胶微丸中的含量准确、快速、重现性好;研究出了替米考星海藻酸钙凝胶微丸稳定的储存条件。本试验对提高替米考星的利用度和对其进一步研究开发提供了基础。
Tilmicosin has been used specially for livestock and poultry as antibiotics because it has excellent antibacterial activity and pharmacokinetics characteristics.The curative effect of tilmicosin is certain, significant and safe.But due to its bitter taste it is very difficult to drive animals taking the medicine,in addition tilmicosin is forbidden to be used by intravenous injection.All of these limit the use oftilmicosin.
To explore the optimum preparation technology of tilmicosin calcium alginate gel micro pills,the effect factors,such as the concentrations of sodium alginate and calcium chloride,the time of calcification,drip system parameters and the ratio of sodium alginate and tilmicosin were investigated by single-factor test, using drug loading and encapsulation efficiency as indicators.In addition,the best preparation technology was obtained by the four factor-three level orthogonal experiment with the ratio of sodium alginate and tilmicosin,the concentration of sodium alginate and calcium chloride and the time of calcification.The optimum preparation technology is afforded as follows:2%concentration of sodium alginate,2% concentration of calcium chloride,1 hour time of calcification,8cm distance between the solution surface and needle,30 drops/min speed of dripping,No 12 needle type and 1:5 ratio of sodium alginate and Tilmicosin. Under the optimum conditions,the gel micro pills enclosing Tilmicosin can be obtained,which have better appearance,higher drug loading and encapsulation and lower bitterness.
RP-HPLC method was used to assay quantificationly the content of Tilmicosin containd in gel micro pills. Mobile phase was composed with water-acetonitrile-tetrahydrofuran-Phosphate Dibutylamine(805:115:55:25) buffer solution,the column temperature was about 30℃,the wavelength of detection was 280 nm,the sample injection volumn is 10μL.A good linear relationship were found between the peak area sum(Y) of cis and trans tilmicosins and their concentration(X).In 0.25-0.85mg/ml concentration range of tilmicosins,the regression equation is Y=565252X+53876,R~2=0.9993.The content of tilmicosins contained in calcium alginate gel micro pills was 76.13%.
To explore the stability of micro pills under different conditions,influencing factors were tested, including high-temperature test,humidity test,the pilot light,accelerated test and long-term test.The experimental results showed that micro pills are sensitive to light and the color,cluster and content of tilmicosin were changed seriously after micro pills were heated on 60℃for 24 hours.But,the micro pills keep a good stability at room temperature.All of these evidence indicated that the micro pills can not be stored under higher temperature.
In the thesis,the preparation technology of tilmicosin calcium alginate gel micro pills explored is perfect and stable with better appearance,higher drug -loading rate and lower bitterness.RP-HPLC method to assay quantificationly the content of tilmicosin containd in gel micro pills is accuate and rapid with good repeatability.At the same time,favorable storage conditions of tilmicosin calcium alginate gel micro pills is obtained.Micro pills should be preserved at lower temperature and away from the light.This thesis supply a beneficial base for tilmicosin researching and exploring futher.
引文
[1]刘小艳,杨利军,邱电,等.畜禽专用抗生素替米考星.动物医学进展,2007,28(4):101-104
[2]马萍,孙淑英.一种新的缓释载体-海藻酸钙凝胶小球的研究概况.国外医药-合成药、生化药制剂分册,1998,19(3):190-192
[3]陈蕾,罗志刚,何小维.海藻酸钠在医学工程上的应用研究进展.医疗卫生装备,2008,29(9):33-35
[4]马萍,孙淑英,辛艳茹,等.硝苯地平海藻酸钙凝胶缓释微丸处方及工艺的优化.沈阳药科大学学报,2003,23(6):405-408
[5]马燕,李卫中,陈大为.粉防己碱海藻酸钙缓释凝胶微丸的制备.中药材,2008,38(1):131-134
[6]吕慧侠,周建平,戴影秋,等.海藻酸钙掩味微囊的制备.中国药科大学学报,2007,38(2):125-128
[7]闫春芝,赵红彦,秦朝英.高效液相色谱法测定替米考星含量.中国兽药杂志,2006,40(8):26-27
[8]The United States Pharmacopeial Convention.United States Pharmacopeia24.000.1661-1662
[9]中华人民共和国兽药典委员会编.中华人民共和国兽药典一部,北京,中国农业出版社,2005:23-25,32-36,95,222-229
[10]中华人民共和国药典委员会编.中华人民共和国药典一部,北京,化学工业出版社,2005:10-11,28-30,71-72
[11]张彦青,解军波,戚务勤,等.灯盏花素海藻酸钙凝胶小球的制备.中草药,2006,37(9):1333-1335
[12]孙淑萍,李高沈灵佳.萘普生海藻酸钙凝胶微丸中萘普生的含量测定.中国药师,2006,9(6):519-520
[13]刘家稳,唐芳,冯裕,等.高湿和高温对齐墩果酸滴丸稳定性的影响.中南药学,2006,4(2):89-90
[14]孙淑萍,李高,沈灵佳,等.萘普生海藻酸钙凝胶微丸的制备及体外释药.中华医院药学杂志,2007,27(4):485-487
[15]KuriharaK,KatsuragiY,MatsuokaI,etal.Receptormechanisms of bitter substances[J],Physiol Behaw,1994,56(6):1125-1132.
[1]郑俊民,平其能.药用高分子材料学.北京:中国医药科技出版社.2000:71-80.
[2]钱风云,傅得贤,欧阳藩.中国海洋药物.2003,91(1):55-56.
[3]周海钧.中国药学杂志.北京:中国药学会,2000:221-231.
[5]王津,李柱来,陈莉敏,等.壳聚糖-海藻酸钠布洛芬缓释微球的制备工艺及性能.福建医科大学学报,2008,42(1):56-58
[6]陈蕾,罗志刚,何小维.海藻酸钠在医学工程上的应用研究进展.医疗卫生装备,2008,29(9):33-35
[7]马萍,祝力,梁冬梅,等.海藻酸钙凝胶微球的制备和pH依赖性溶胀.中国海洋药物,2003,22(5):36-39
[8]马萍,孙淑英,辛艳茹,等.硝苯地平海藻酸钙凝胶缓释微丸处方及工艺的优化.沈阳药科大学学报,2003,23(6):405-408
[9]Naoki Segi.Chem Plarm Bull,1989,37:3092
[10]张彦青,解军波,戚务勤,等.灯盏花素海藻酸钙凝胶小球的制备.中草药,2006,37(9):1333-1335
[11]马萍,孙淑英.一种新的缓释载体-海藻酸钙凝胶小球的研究概况.国外医药-合成药、生化药制剂分册,1998,19(3):190-192
[12]于飞千,刘臻,潘卫三.药物制剂中苦味掩盖方法的研究进展.中国新药杂志,2007,16(15):1171-1174
[13]毕殿洲.药剂学[M].第4版.北京:人民卫生出版社,1999:202-203
[14]刘东春,崔福德,福森义信,等.流化床包衣法制备100μm级马来酸氯苯那敏掩味微囊.沈阳药科大学学报,2006,23f3):129-135.
[15]朱海涛,陈吉炎,黄良永,等.奥硝唑β2环糊精包合物的制备及其质量控制.中国药房,2005,16(17):1305-1306.
[16]宋韵梅,平其能,张志燕.曲马多药物树脂速释混悬剂的研制.中国药科大学学报,2000,31(1);18-23.
[17]黄胜炎.苦味阻滞剂.上海医药,2004,25(12):542-546.
[18]吕慧侠,周建平,戴影秋,等.海藻酸钙掩味微囊的制备.中国药科大学学报,2007,38(2):125-128