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摘 要 建立了用高效液相色谱测定酮洛芬贴片含量的方法并对其进行了稳定性研究.固定相:HYPERSIL ODS C18.流动相:甲醇-0.05 mol.L-1磷酸二氢钾(55∶45).检测波长:258 nm.酮洛芬浓度在6.696~33.48 μg.mL-1范围内,线性关系良好,r=0.9993,平均回收率95.17%,RSD=0.75%,日内、日间差RSD均小于1.5%.本法适用于酮洛芬贴片的质量控制和稳定性研究. Determination of Ketoprofen in a Patch by HPLC and the Study on Its Stability Zhang Zhongyuan,Yue Li,Zhang Qingping,Tong Zhiqing,Wang Yufeng Abstract A HPLC method was established for the determination of ketoprofen in a patch and the study on its stability.The HYPERSIL ODS C18 column was used.The mobile phase consisted of methanol-water(55∶45)with 0.05 mol*L-1 potassium dihydrogen phosphate.The detective wavelength was 258 nm.The linear range was from 6.696 to 33.48 μg*mL-1,r=0.999 3.The average recovery was 95.17% with RSD=0.75%.The method can be used in the determination of ketoprofen in a patch and the study on its stability. 酮洛芬是一种非甾体抗炎镇痛药,《中华人民共和国药典》已收载其原料药和肠溶胶囊剂,本品口服后有胃肠道刺激等副作用,将其制成透皮给药制剂可避免上述副作用,并可提高病灶部位组织中的药物浓度,增强其抗炎镇痛作用.制剂中酮洛芬的含量测定方法已报导的有中和滴定法及紫外分光光度法,但由于酮洛芬贴片中成分复杂、干扰物质多而无法采用.本文建立了酮洛芬贴片含量测定的HPLC法,进行了该贴片对光、热的稳定性加速试验,预测了其有效期. 1 实验材料 810液相色谱系统(美国WATERS公司);甲醇(色谱纯);磷酸二氢钾及环己烷为分析纯;磷酸盐缓冲液(pH7.2,按中国药典配制);酮洛芬(西南合成药厂,含量99.93%);对甲氧基苯甲酸(分析纯);酮洛芬贴片(自制,批号:970521、970522、970523);空白贴片(自制,除不含酮洛芬外其余成分及制备方法与酮洛芬贴片相同);酮洛芬标准溶液(精取经五氧化二磷60℃减压干燥至恒重的酮洛芬适量,用流动相溶解并稀释,使含酮洛芬约35μg.mL-1);内标溶液(精取对甲氧基苯甲酸适量,用流动相溶解并稀释,使含对甲氧基苯甲酸约20 μg.mL-1). 2 方法与结果 2.1 样品处理
Fig.1 Chromatograms of a blank patch 2.3.2 贴片中分解产物的干扰
Fig.2 Chromatograms of the ketoprofen patch with interstandard
Fig.3 Chromatograms of the ketoprofen patch with interstandard under optical tests 从以上试验可看出,本法专属性强,适用于该制剂的质量控制和稳定性研究. Tab.1 Recoveries(n=5) |
Amount added /μg.mL-1 |
Amount founded /μg.mL-1 |
Recoveries /% |
RSD /% |
0.2678 | 0.2549 | 95.19 | 1.06 |
0.4018 | 0.3801 | 94.60 | 0.49 |
0.5357 | 0.5128 | 95.73 | 0.71 |
Average recovery:95.17%;RSD:0.75% 2.6 方法精密度试验 选择线性关系试验中5个浓度样品中的任意3个浓度样品进行日内及日间的测定结果差异考察(n=3).结果为,日内:RSD=1.01%,日间:RSD=1.39%. 2.7 制剂含量测定 取3个批号的样品,照“2.1.1”项操作,进样,结果见表2. Tab.2 Results of quantitation(n=3) |
Batch No | Actual amount/% | RSD/% |
970521 |
102.9 |
0.52 |
970522 | 97.83 | 0.45 |
970523 | 98.75 | 0.28 |
2.8 光照实验 取样品置光照度为3 000 Lux处,分别于放置样品后0、1、3、6、10 d取出,观察性状变化,用HPLC法测定含量,操作同2.1.1项,结果见表3.本品经光照后不稳定,易变色,含量降低,应避光保存. Tab.3 Results of optical tests |
Time /d |
970521 | 970522 | 970523 | |||
Content /% |
Color** | Content /% |
Color | Content /% |
Color | |
1 |
98.86 | - | 99.14 | - | 98.44 | - |
3 | 96.34 | + | 96.07 | + | 97.53 | + |
6 | 95.80 | + | 93.42 | + | 95.25 | + |
10 | 91.91 | + | 92.19 | ++ | 93.01 | ++ |
Content/%=(Amount of time t/Amount of time 0)×100%; Color:“-”No change,“+”Yellow,“++”Dark yellow
2.9 制剂稳定性预测〔2〕 Tab.4 V0(%.d-1)and color change |
Temp /℃ |
Time /d |
970521 | 970522 | 970523 | |||||||||
V0 | Color | V0 | Color | <V0 | Color | <||||||||
/%.d-1 | /%.d-1 | /%.d-1 | |||||||||||
70 | 1 | 0.380 | - | 0.321 | - | 0.456 | - | ||||||
65 | 2 | 0.302 | - | 0.260 | - | 0.335 | - | ||||||
60 | 3 | 0.234 | - | 0.195 | - | 0.349 | - | ||||||
55 | 4 | 0.165 | - | 0.159 | - | 0.220 | - | ||||||
50 | 5 | 0.097 | - | 0.102 | - | 0.167 | - | ||||||
45 | 6 | 0.055 | - | 0.041 | - | 0.078 | - | ||||||
40 | 7 | 0.014 | - | 0.020 | - | 0.017 | - |
-No color change 取室温为25℃(T=298)代入logV0-T-1的线性回归方程中,求出V0(25),再将V0(25)代入公式t0.9=0.1C0/V0(25),得t0.9(见表5).酮洛芬贴片对热较稳定,降解速率较小,色泽也未改变,室温贮存的有效期(t0.9)均在2年以上,可暂定本品在室温避光条件下的贮存期为2年. Tab.5 Regressive equation of logV0-T-1 and storage life |
Batch | A | B | r | V0(25)/%.d-1 | t0.9/a |
970521 | 13.611 5 | -4 763.79 | 0.955 8 | 4.223 1×10-3 | 3.2 |
970522 | 12.016 3 | -4 249.69 | 0.965 7 | 5.696 3×10-3 | 2.3 |
970523 | 13.091 2 | -4 551.40 | 0.919 0 | 6.577 3×10-3 | 2.0 |
3 讨论
a.中国药典1995年版和英国药典1988年版中酮洛芬原料及制剂的含量测定方法均为中和滴定法,也有报道采用紫外分光光度法进行制剂分析〔3〕,但经实验均不能消除本贴片中辅料和降解产物的干扰.作者参考了葛庆华等〔1〕、陈琰等〔4〕的方法所建立的HPLC法具有较好的专属性,可用于该贴片的含量测定及稳定性研究.
参考文献 收稿日期:1999-08-30 |