点击显示 收起
摘 要 采用高效液相色谱法,对氟马西尼及其合成过程中产生的中间产物以及有关杂质进行分离测定.结果表明:对照品进样量在2~10
μg范围内有良好线性关系,且分离时间短,重复性好,含量测定结果令人满意. Determination of the Content of Flumazenil by HPLC Wang Yuchuan, Sun
Xiuyan, Wang Liqun, Cheng Maosheng Abstract In this
paper,flumazenil,intermediates and impurities were separated and
determined quantitatively by highperformance liquid chromatography with
the C18 column.The mobile phase was acetonitrile and
water(V∶V=22∶78).The linear ranges of the standard sample were 2~10 μg.By
the method,the short retention time was readily achieved with satisfactary
results. 氟马西尼(flumazenil)为苯二氮受体选择性拮抗剂,临床上用于终止由苯二氮类药物诱导维持的全身麻醉,以及该类药物中毒的急救.沈阳药科大学合成室经大量实验,筛选出氟马西尼的最佳合成路线.作者采用HPLC法,以十八烷基硅烷键合硅胶为填充剂,乙腈-水(V∶V=22∶78)为流动相,氟羟肟作内标物测定氟马西尼的含量,在该色谱条件下,与主峰及杂质峰均能很好分离,且分离时间短,重复性好,含量测定结果令人满意. Tab.1 The results of elementary analysis% |
Element | C | H | O | N | F |
Theory value | 59.40 | 4.65 | 13.85 | 15.85 | 6.26 |
Practical value | 59.50 | 4.85 | 13.73 | 15.80 | 6.15 |
在上述色谱条件下测得氟马西尼对照品的纯度为99.7%,RSD为0.96%(n=5).
3 方法与结果 Tab.2 The linear relationship |
No. | Amount of standard
sample /μg |
Ai | As | Ai/As | Average value
of Ai/As |
1 | 1.904 | 6733036 | 21828601 | 0.308 4 | 0.308 5 |
6751434 | 21879520 | 0.308 6 | |||
2 | 3.808 | 13340769 | 22026443 | 0.605 7 | 0.606 5 |
13494769 | 22222792 | 0.607 2 | |||
3 | 5.712 | 20306802 | 22266642 | 0.912 0 | 0.912 3 |
20238642 | 22176817 | 0.912 6 | |||
4 | 7.616 | 25342712 | 20991629 | 1.207 3 | 1.208 0 |
25098347 | 20764397 | 1.208 7 | |||
5 | 9.520 | 33302574 | 22122456 | 1.505 8 | 1.506 0 |
33152533 | 22011101 | 1.506 2 |
3.2 供试品溶液的制备 取氟马西尼样品约100 mg,精密称定,置100 mL量瓶中,以甲醇溶解并稀释至刻度,摇匀.精密取该溶液3 mL,置10 mL量瓶中,以甲醇稀释至刻度,摇匀,作为供试品溶液. 3.3 稳定性考察 精密吸取同一供试品溶液,按0,2,4,6,8,10 h时间间隔,分别测定供试品峰面积(Ai)和内标物峰面积(As)值,测定结果见表3. 实验结果表明:供试品溶液在制备后10 h内测定,色谱峰面积无明显变化,说明在此时间内测定,供试品化学性质稳定. Tab.3 The results of the stability observation |
t/h | Ai | As | Ai/As | Sample content/g*L-1 | RSD/% |
0 |
19602959 | 22236161 | 0.881 6 | 0.275 2 | |
2 | 19608817 | 22232634 | 0.882 3 | 0.275 4 | |
4 | 19356163 | 21991813 | 0.880 2 | 0.274 7 | 0.2 |
6 | 19468412 | 22088780 | 0.881 4 | 0.275 1 | |
8 | 19535530 | 22132293 | 0.882 6 | 0.275 5 | |
10 | 19413831 | 22113877 | 0.877 9 | 0.274 0 |
3.4 重现性考察
制备6份同一批号的供试品溶液,分别按上述色谱条件测定,测定结果见表4. Tab.4 The results of precision tests |
No. | Ai | As | Ai/As | RSD/% |
1 | 19356163 | 21991813 | 0.880 2 | |
2 | 19468412 | 22088780 | 0.881 4 | |
3 | 19535530 | 22132293 | 0.882 7 | 0.87 |
4 | 19413931 | 22113877 | 0.887 9 | |
5 | 19836764 | 22058207 | 0.899 3 | |
6 | 19495236 | 22059393 | 0.883 8 |
3.5 样品的测定 测定3批样品,测定结果见表5.氟马西尼对照品的浓度为0.952 g/L. Tab.5 The measuremental results of the sample contents |
No. | Sample
concentration /g。L-1 |
Ai | As | Ai/As | Average value of Ai/As |
Content/% |
1 | 0.989 | 20939724 | 22173321 | 0.944 4 | 0.943 5 | 99.2 |
21035001 | 22316770 | 0.942 6 | ||||
2 | 0.952 | 19836764 | 22058207 | 0.899 3 | 0.898 4 | 98.1 |
19796316 | 22057100 | 0.897 5 | ||||
3 | 0.968 | 20251277 | 22070245 | 0.917 6 | 0.9166 | 98.5 |
20246904 | 22111612 | 0.915 7 |
Fig.1 Chromatograms of flumazenil and its intermediates 4 讨论 4.1 中间产物的分离 王玉川(银川市第二人民医院,银川 750000) 参考文献 1,中华人民共和国卫生部药典委员会.中华人民共和国药典:二部.北京:人民卫生出版社,1995.附录25~26
|