时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
7 | 90 | 10 |
11 | 90 | 10 |
含量测定方法学验证结果概要
项目 | 验证结果 |
波长选择 | 参照有关物质项下,波长选定为225nm |
流动相选择 | 参照有关物质项流动相组分,流速为1.2ml/min |
进样精密度试验 | 连续测量6次,主峰保留时间和峰面积RSD值均小于2.0% |
线性关系试验 | 阿莫西林的浓度在154.3μg/ml~360.1g/ml,直线方程为y=53931x+117907,R2=0.9995,相关系数r=0.9997,截距为117907,小于100%浓度溶液阿莫西林峰面积的±2.0%(282392),表明线性关系试验符合要求; 双氯西林钠的浓度在79.1μg/ml~184.5g/ml,直线方程为y=85091x+684344,R2=0.9990,相关系数r=0.9995,截距为684344。 |
溶液稳定性试验 | 室温条件下置8个小时,测得阿莫西林和双氯西林钠的主峰面积的RSD%值均小于2.0% |
回收率试验 | 阿莫西林平均回收率为101.3%,在98.0%~102.0%范围内,n=9,RSD为0.67%,小于2.0%,回收率符合含量测定方法验证要求。 双氯西林钠平均回收率为100.6%,在98.0%~102.0%范围内,n=9,RSD为1.38%,小于2.0%,回收率符合含量测定方法验证要求。 |
重复性试验 | 试验结果为99.02%~102.17%,RSD%值均小于2.0%,符合重复性试验要求 |
含量测定结果 | 上市品和自制品均符合规定 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
7 | 90 | 10 |
11 | 90 | 10 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.527 | 0.00000 | 32655 | 0.1088 | 1.784 | -- | 1064.449 |
2 | 4.379 | 0.00000 | 60908 | 0.2030 | 1.344 | 11.664 | 3370.822 |
3 | 6.648 | 0.00000 | 16679096 | 55.5970 | 1.011 | 6.348 | 4138.199 |
4 | 8.447 | 0.00000 | 53214 | 0.1774 | 0.838 | 3.540 | 3138.427 |
5 | 13.147 | 0.00000 | 30827 | 0.1028 | 0.747 | 6.976 | 4993.018 |
6 | 15.711 | 0.00000 | 65280 | 0.2176 | 0.947 | 2.765 | 3200.106 |
7 | 20.718 | 0.00000 | 13077997 | 43.5934 | 0.936 | 4.365 | 4907.052 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
6 | 90 | 10 |
15 | 90 | 10 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.459 | 0.00000 | 56751 | 0.1852 | 1.083 | -- | 569.474 |
2 | 1.846 | 0.00000 | 64775 | 0.2114 | 1.793 | 1.740 | 1358.467 |
3 | 2.222 | 0.00000 | 7530 | 0.0246 | -- | 1.442 | 765.105 |
4 | 2.362 | 0.00000 | 32845 | 0.1072 | -- | 0.512 | 1749.452 |
5 | 2.962 | 0.00000 | 50698 | 0.1654 | 1.144 | 2.745 | 3140.381 |
6 | 3.788 | 0.00000 | 16353808 | 53.3652 | 1.103 | 3.547 | 3551.516 |
7 | 4.292 | 0.00000 | 2570 | 0.0084 | 1.375 | 2.158 | 6489.602 |
8 | 4.596 | 0.00000 | 11024 | 0.0360 | 1.106 | 1.356 | 6071.005 |
9 | 5.605 | 0.00000 | 262795 | 0.8575 | 0.722 | 2.864 | 2291.990 |
10 | 6.629 | 0.00000 | 15761 | 0.0514 | 0.804 | 2.711 | 8523.512 |
11 | 6.993 | 0.00000 | 69584 | 0.2271 | 1.034 | 1.109 | 5737.622 |
12 | 7.605 | 0.00000 | 21020 | 0.0686 | 0.916 | 1.847 | 10809.034 |
13 | 8.152 | 0.00000 | 21792 | 0.0711 | 0.824 | 1.376 | 4191.092 |
14 | 10.215 | 0.00000 | 13492960 | 44.0298 | 0.958 | 4.257 | 7715.397 |
15 | 11.147 | 0.00000 | 131177 | 0.4281 | 0.959 | 1.930 | 7927.547 |
16 | 11.793 | 0.00000 | 32746 | 0.1069 | 1.081 | 1.286 | 8767.838 |
17 | 12.428 | 0.00000 | 8921 | 0.0291 | 1.040 | 1.336 | 12356.013 |
18 | 13.014 | 0.00000 | 8309 | 0.0271 | 1.161 | 1.168 | 8749.285 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 5 |
5 | 85 | 15 |
6 | 90 | 5 |
15 | 90 | 5 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.963 | 0.00000 | 71698 | 0.2403 | 1.143 | -- | 1381.826 |
2 | 2.731 | 0.00000 | 22631 | 0.0759 | 1.189 | 3.710 | 2904.008 |
3 | 3.443 | 0.00000 | 54394 | 0.1823 | 0.898 | 3.242 | 3394.891 |
4 | 4.753 | 0.00000 | 16353295 | 54.8183 | 1.068 | 4.861 | 3944.750 |
5 | 5.142 | 0.00000 | 35224 | 0.1181 | 1.837 | 1.413 | 6920.616 |
6 | 7.427 | 0.00000 | 31653 | 0.1061 | 0.884 | 8.457 | 10275.546 |
7 | 8.144 | 0.00000 | 227994 | 0.7643 | 0.747 | 1.870 | 4717.508 |
8 | 8.847 | 0.00000 | 121630 | 0.4077 | 0.779 | 1.656 | 8906.010 |
9 | 9.270 | 0.00000 | 38235 | 0.1282 | 3.251 | 0.658 | 1655.368 |
10 | 12.157 | 0.00000 | 12875049 | 43.1588 | 0.956 | 3.190 | 2932.138 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 5 |
5 | 85 | 15 |
6 | 90 | 5 |
15 | 90 | 5 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.654 | 0.00000 | 66056 | 0.2658 | 1.103 | -- | 1100.578 |
2 | 2.295 | 0.00000 | 20069 | 0.0808 | 1.158 | 3.375 | 2586.084 |
3 | 2.917 | 0.00000 | 45432 | 0.1828 | 0.857 | 3.191 | 3094.039 |
4 | 4.034 | 0.00000 | 13553808 | 54.5481 | 1.060 | 4.648 | 3551.711 |
5 | 4.399 | 0.00000 | 140841 | 0.5668 | -- | 1.051 | 1737.970 |
6 | 6.281 | 0.00000 | 28612 | 0.1152 | 0.964 | 5.383 | 8201.704 |
7 | 7.046 | 0.00000 | 207751 | 0.8361 | 0.720 | 2.185 | 4446.737 |
8 | 7.640 | 0.00000 | 77019 | 0.3100 | 1.021 | 1.622 | 9703.165 |
9 | 9.811 | 0.00000 | 10535378 | 42.4003 | 1.023 | 4.341 | 3234.582 |
10 | 12.582 | 0.00000 | 172467 | 0.6941 | 0.915 | 3.372 | 2783.828 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
6 | 90 | 10 |
15 | 90 | 10 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.535 | 0.00000 | 74235 | 0.2803 | 1.633 | -- | 1277.766 |
2 | 1.966 | 0.00000 | 33932 | 0.1281 | -- | 2.272 | 1440.322 |
3 | 2.463 | 0.00000 | 42410 | 0.1601 | 1.135 | 2.535 | 2851.813 |
4 | 3.146 | 0.00000 | 13691792 | 51.6941 | 1.097 | 3.363 | 3219.236 |
5 | 3.832 | 0.00000 | 24103 | 0.0910 | 1.058 | 3.140 | 5061.970 |
6 | 4.645 | 0.00000 | 315595 | 1.1915 | 0.790 | 2.786 | 2549.413 |
7 | 5.540 | 0.00000 | 38306 | 0.1446 | 0.854 | 2.846 | 7190.080 |
8 | 5.864 | 0.00000 | 81436 | 0.3075 | 1.077 | 1.118 | 5433.266 |
9 | 6.847 | 0.00000 | 55119 | 0.2081 | 0.858 | 2.433 | 3132.324 |
10 | 8.644 | 0.00000 | 10751027 | 40.5911 | 0.965 | 3.937 | 6655.264 |
11 | 9.515 | 0.00000 | 175822 | 0.6638 | 0.995 | 1.839 | 5285.921 |
12 | 10.268 | 0.00000 | 73123 | 0.2761 | 1.041 | 1.340 | 4690.931 |
13 | 15.921 | 0.00000 | 66541 | 0.2512 | 0.680 | 6.522 | 3155.731 |
14 | 18.278 | 0.00000 | 1062743 | 4.0124 | 1.917 | 1.162 | 624.839 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
6 | 90 | 10 |
12 | 90 | 10 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.212 | 0.00000 | 65163 | 0.2553 | 0.893 | -- | 476.924 |
2 | 1.544 | 0.00000 | 84040 | 0.3292 | 1.556 | 1.668 | 1241.130 |
3 | 1.974 | 0.00000 | 35690 | 0.1398 | -- | 2.233 | 1407.655 |
4 | 2.471 | 0.00000 | 46103 | 0.1806 | 1.116 | 2.493 | 2773.460 |
5 | 3.160 | 0.00000 | 13745616 | 53.8510 | 1.099 | 3.362 | 3226.914 |
6 | 3.838 | 0.00000 | 29088 | 0.1140 | 1.048 | 3.063 | 4877.921 |
7 | 4.698 | 0.00000 | 344543 | 1.3498 | 0.775 | 2.936 | 2633.628 |
8 | 5.539 | 0.00000 | 50749 | 0.1988 | 0.872 | 2.647 | 6769.839 |
9 | 5.887 | 0.00000 | 105164 | 0.4120 | 1.062 | 1.156 | 4986.538 |
10 | 6.878 | 0.00000 | 73013 | 0.2860 | 0.840 | 2.356 | 2940.947 |
11 | 8.654 | 0.00000 | 10647600 | 41.7140 | 0.964 | 3.811 | 6640.418 |
12 | 9.528 | 0.00000 | 189786 | 0.7435 | 0.998 | 1.840 | 5265.736 |
13 | 10.281 | 0.00000 | 86236 | 0.3378 | 1.040 | 1.333 | 4611.522 |
14 | 11.015 | 0.00000 | 22462 | 0.0880 | 1.076 | 1.262 | 6225.514 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
7 | 90 | 10 |
12 | 90 | 10 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 0.357 | 0.00000 | 24153 | 0.0948 | 1.245 | -- | 3.366 |
2 | 1.216 | 0.00000 | 59810 | 0.2346 | 1.125 | 1.736 | 531.570 |
3 | 1.545 | 0.00000 | 85015 | 0.3335 | 1.502 | 1.715 | 1267.735 |
4 | 1.981 | 0.00000 | 39243 | 0.1539 | -- | 2.225 | 1325.908 |
5 | 2.460 | 0.00000 | 106446 | 0.4176 | -- | 2.105 | 1710.607 |
6 | 3.132 | 0.00000 | 13767653 | 54.0105 | 1.099 | 2.927 | 3205.622 |
7 | 3.843 | 0.00000 | 28933 | 0.1135 | 1.103 | 3.263 | 5136.312 |
8 | 4.601 | 0.00000 | 323063 | 1.2674 | 0.867 | 2.680 | 2745.529 |
9 | 5.555 | 0.00000 | 53699 | 0.2107 | 0.885 | 3.095 | 7020.235 |
10 | 5.868 | 0.00000 | 79747 | 0.3128 | 1.122 | 1.101 | 6021.822 |
11 | 6.814 | 0.00000 | 66373 | 0.2604 | 0.890 | 2.443 | 3334.338 |
12 | 8.678 | 0.00000 | 10544441 | 41.3658 | 0.963 | 4.152 | 6637.561 |
13 | 9.527 | 0.00000 | 197288 | 0.7740 | 0.988 | 1.788 | 5303.372 |
14 | 10.255 | 0.00000 | 90469 | 0.3549 | 1.072 | 1.294 | 4640.592 |
15 | 11.007 | 0.00000 | 24384 | 0.0957 | 1.085 | 1.288 | 6086.682 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
7 | 90 | 10 |
12 | 90 | 10 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 0.581 | 0.00000 | 38286 | 0.0226 | 0.805 | -- | 9.917 |
2 | 0.988 | 0.00000 | 74946 | 0.0442 | 1.696 | 0.830 | 270.960 |
3 | 1.487 | 0.00000 | 773082 | 0.4564 | 0.776 | 2.626 | 1795.658 |
4 | 1.965 | 0.00000 | 555965 | 0.3282 | -- | 2.899 | 1726.814 |
5 | 2.542 | 0.00000 | 84265404 | 49.7428 | 0.860 | 2.779 | 2020.394 |
6 | 3.114 | 0.00000 | 82014 | 0.0484 | -- | 2.755 | 4320.435 |
7 | 3.724 | 0.00000 | 3929502 | 2.3196 | 0.906 | 2.277 | 1850.921 |
8 | 4.530 | 0.00000 | 217118 | 0.1282 | -- | 1.867 | 1228.697 |
9 | 4.799 | 0.00000 | 377030 | 0.2226 | -- | 0.479 | 1007.064 |
10 | 5.236 | 0.00000 | 166356 | 0.0982 | -- | 0.939 | 4112.661 |
11 | 5.538 | 0.00000 | 641704 | 0.3788 | -- | 0.853 | 3348.896 |
12 | 6.278 | 0.00000 | 62892 | 0.0371 | 0.845 | 1.778 | 3121.250 |
13 | 7.190 | 0.00000 | 76883552 | 45.3853 | 0.970 | 2.175 | 5473.031 |
14 | 7.863 | 0.00000 | 1240644 | 0.7324 | 0.992 | 1.695 | 6022.781 |
15 | 8.378 | 0.00000 | 24514 | 0.0145 | 1.133 | 1.356 | 8930.174 |
16 | 8.858 | 0.00000 | 39248 | 0.0232 | 1.334 | 1.110 | 4817.893 |
17 | 9.483 | 0.00000 | 29794 | 0.0176 | 1.431 | 1.287 | 6771.330 |
峰# | 保留时间 | 浓度 | 面积 | 面积% | 拖尾因子 | 分离度 | 理论塔板# |
1 | 1.231 | 0.00000 | 76271 | 0.3941 | 1.499 | -- | 1085.221 |
2 | 1.491 | 0.00000 | 21444 | 0.1108 | -- | 1.260 | 514.264 |
3 | 1.593 | 0.00000 | 29310 | 0.1514 | -- | 0.387 | 586.781 |
4 | 1.973 | 0.00000 | 34767 | 0.1796 | 1.209 | 1.785 | 2337.839 |
5 | 2.529 | 0.00000 | 10639693 | 54.9718 | 1.102 | 3.095 | 2671.539 |
6 | 3.115 | 0.00000 | 54780 | 0.2830 | 1.037 | 2.990 | 4023.005 |
7 | 3.751 | 0.00000 | 501931 | 2.5933 | 0.915 | 2.248 | 1651.116 |
8 | 4.519 | 0.00000 | 113110 | 0.5844 | 0.916 | 2.482 | 5225.541 |
9 | 4.819 | 0.00000 | 92677 | 0.4788 | 1.153 | 1.144 | 4961.773 |
10 | 5.631 | 0.00000 | 129885 | 0.6711 | 0.895 | 2.263 | 2574.263 |
11 | 7.200 | 0.00000 | 7213868 | 37.2717 | 0.960 | 3.820 | 5815.185 |
12 | 7.863 | 0.00000 | 246286 | 1.2725 | 0.933 | 1.731 | 6572.042 |
13 | 8.353 | 0.00000 | 160155 | 0.8275 | 1.046 | 1.221 | 6474.555 |
14 | 8.817 | 0.00000 | 40642 | 0.2100 | 1.320 | 1.138 | 7781.493 |
精密度试验结果
进针次数 | 1 | 2 | 3 | 4 | 5 | 6 | RSD% |
阿莫西林保留时间(min) | 3.114 | 3.086 | 3.094 | 3.091 | 3.115 | 3.118 | 0.5 |
阿莫西林峰面积 | 14299619 | 14184091 | 14162667 | 14183664 | 14096544 | 14124837 | 0.5 |
双氯西林钠保留时间(min) | 8.698 | 8.705 | 8.704 | 8.700 | 8.710 | 8.695 | 0.1 |
双氯西林钠峰面积 | 11657316 | 11623882 | 11529543 | 11454990 | 11357474 | 11268283 | 1.3 |
阿莫西林线性关系试验结果
浓度(%) | 60% | 80% | 100% | 120% | 140% |
浓度(μg/ml) | 154.3 | 205.7 | 257.2 | 308.7 | 360.1 |
峰面积(A) | 8353611 | 11219467 | 14119578 | 16824904 | 19427332 |
双氯西林钠线性关系试验结果
浓度(%) | 60% | 80% | 100% | 120% | 140% |
浓度(μg/ml) | 79.1 | 105.4 | 131.8 | 158.1 | 184.5 |
峰面积(A) | 7295163 | 9704301 | 12048197 | 14164961 | 16275670 |
溶液中阿莫西林稳定性试验结果
放置时间(h)/峰面积 | ||||||
0 | 2 | 4 | 6 | 8 | 平均值 | RSD |
13938821 | 13900520 | 13897546 | 13919668 | 13880679 | 13907447 | 0.2% |
溶液中双氯西林钠稳定性试验结果
放置时间(h)/峰面积 | ||||||
0 | 2 | 4 | 6 | 8 | 平均值 | RSD |
11273568 | 11278077 | 11275616 | 11279914 | 11236201 | 11268675 | 0.2% |
阿莫西林回收率试验测定结果
标示量 | 加入量(g) | 测得量(g) | 回收率(%) | 平均值(%) | RSD(%) |
80% | 0.0586 | 0.0519 | 101.93 | 101.3 | 0.67 |
0.0588 | 0.0519 | 101.48 | |||
0.0586 | 0.0517 | 101.52 | |||
100% | 0.0728 | 0.0643 | 101.62 | ||
0.0732 | 0.0648 | 101.91 | |||
0.0729 | 0.0646 | 101.94 | |||
120% | 0.0872 | 0.0761 | 100.49 | ||
0.0874 | 0.0763 | 100.44 | |||
0.0874 | 0.0762 | 100.37 |
双氯西林钠回收率试验测定结果
标示量 | 加入量(g) | 测得量(g) | 回收率(%) | 平均值(%) | RSD(%) |
80% | 0.0276 | 0.0262 | 100.20 | 100.6 | 1.38 |
0.0273 | 0.0256 | 99.03 | |||
0.0271 | 0.0251 | 97.85 | |||
100% | 0.0358 | 0.0343 | 100.93 | ||
0.0359 | 0.0345 | 101.42 | |||
0.0358 | 0.0344 | 101.41 | |||
120% | 0.0426 | 0.0412 | 102.10 | ||
0.0428 | 0.0411 | 101.27 | |||
0.0428 | 0.0412 | 101.60 |
重复性试验结果(%)
成分 | 1 | 2 | 3 | 4 | 5 | 6 | 平均值 | RSD |
阿莫西林 | 99.21 | 99.41 | 99.60 | 101.95 | 102.04 | 102.17 | 100.7 | 1.45 |
双氯西林钠 | 101.53 | 101.50 | 100.83 | 99.39 | 99.20 | 99.02 | 100.2 | 1.17 |
时间(分钟) | 流动相A(%) | 流动相B(%) |
0 | 90 | 10 |
5 | 85 | 15 |
7 | 90 | 10 |
11 | 90 | 10 |