硫醇汞盐紫外分光光度法测定尿液中羟氨苄青霉素的含量
作者:王焕 江晓君 满羽
单位:王焕(鞍钢铁东医院药剂科,鞍山 114002);江晓君(武警辽宁总队医院药剂科,沈阳 110026);满羽(中国医科大学第一临床学院药剂科,沈阳 110001)
关键词:硫醇汞盐紫外分光光度法;羟氨苄青霉素;药代动力学
沈阳药科大学学报00zk08 摘 要 采用硫醇汞盐紫外分光光度法测定尿中羟氨苄青霉素的含量.方法回收率为(99.1±1.25)%,相对标准差为1.26%(n=6),线性范围6~60 μg/mL,日内、日间变异系数小于2.01%和2.94%,结果令人满意.
分类号 R914.1
A Study on the Determination of the Content of Amoxicillin in Urine
, http://www.100md.com
Wang Huan, Jiang Xiaojun, Man Yu
(The Tiedong Hospital of An′gang,Anshan 114002)
Abstract The determination of amoxicillin in urine by UV-spectrophotometry has been developed in this paper. The mean recovery and RSD were(99.1±1.25)% and 1.26%(n=6), respectively. The linearity of amoxicillin was from 6~60 μg.mL-1, and the validations of inter-day and intra-day were less than 2.01% and 2.94%, respectively.
, 百拇医药
Key words UV-spectrophotometry;Amoxicillin;pharmacokinetics
羟氨苄青霉素是广谱抗生素,广泛应用于临床.文献报道羟氨苄青霉素体内测定方法有紫外分光光度法〔1,2〕、高效液相色谱法〔3,4〕、荧光法〔5〕和极谱法〔6〕等.本研究采用硫醇汞盐紫外分光光度法对尿中羟氨苄青霉素的含量进行测定.测得本法平均回收率为(99.1±1.25)%,相对标准差为1.26%(n=6),日内、日间变异系数小于2.01%和2.94%.测定了6名男性健康自愿受试者交叉口服羟氨苄青霉素两种胶囊剂的羟氨苄青霉素药动学参数.
1 材料与方法
1.1 仪器与试剂
日本岛津-260型分光光度计(日本岛津),WFZ 800-D2型分光光度计(北京第二光学仪器厂),pH-25型酸度计,586型计算机.羟氨苄青霉素对照品(中国药品生物制品检定所提供),羟氨苄青霉素胶囊(试验制剂,批号:980809),羟氨苄青霉素胶囊(对照制剂,深圳华南制药有限公司提供,批号:990801).其它试剂均为分析纯.
, 百拇医药
1.2 供试液的配制
羟氨苄青霉素标准液(0.6 mg/mL):取羟氨苄青霉素对照品约0.15 g,精密称定,置250 mL量瓶中,加水溶解并稀释至刻度,摇匀,即得,置冰箱中保存.
硼酸缓冲液(pH 9.0):称取硼酸1.24 g,加水180 mL溶解后,加0.1 mol/L氢氧化钠溶液调pH至9.0,再用水稀释至200 mL,摇匀,即得.
醋酐乙腈溶液:取醋酐1 mL,加乙腈至50 mL,摇匀,即得.
咪唑溶液:取用苯精制过的咪唑8.25 g,加水60 mL溶解后,加盐酸(6 mol/L)8.3 mL,在搅拌下滴加0.27%二氯化汞溶液10 mL,调节pH值6.8±0.05,用水稀释至100 mL,滤过,摇匀,即得.
1.3 测定方法
, http://www.100md.com
精密吸取羟氨苄青霉素标准液或尿液适量,置50 mL量瓶中,加水稀释至刻度,摇匀,精密量取5.0 mL,置25 mL量瓶中,加硼酸缓冲液2.5 mL,醋酐乙腈溶液0.25 mL,放置5 min后,加咪唑溶液至刻度,摇匀,置60℃水浴中,加热30 min,取出、冷却.以水5.0 mL代替尿样,同法操作为空白,于325 nm处测定.
1.4 尿样的采集
男性健康自愿受试者6人,年龄22~24周岁,体重54~65 kg,随机分为两组,每组3人.试验前两周不得服用任何药物,前三天统一进标准餐、标准饮料,晨起空腹交叉服药(试验制剂含羟氨苄青霉素1.5 g,对照制剂含羟氨苄青霉素0.5 g),服药后2 h进标准餐,每次采样后饮温水100 mL,于服药后0.5,1.0,1.5,2.0,3.0,4.0,6.0,8.0 h采集尿液,量取体积后过滤,测定.
2 结果与讨论
, 百拇医药
2.1 测定条件的选择
精密吸取羟氨苄青霉素标准液两份2.5 mL,置50 mL量瓶中,其中一份加入空白滤过尿液1.0 mL,另取1.0 mL空白滤过尿液,按“1.3”项下操作,于250~400 nm间分别绘制吸收光谱,选取325 nm处测定时,空白尿液不干扰.
2.2 标准曲线的绘制
精密吸取羟氨苄青霉素标准液0.5,2.0,2.5,3.0,5.0 mL,分置50 mL量瓶中,按“1.3”项下操作,得回归方程为A=0.009 487+0.065 28C,r=0.999 2,线性范围为6~60 μg/mL,每隔30 min测定一次,结果表明:吸收度在2 h内不变.
2.3 回收率试验
取1.0 mL空白滤过尿液6份,分别精密加入羟氨苄青霉素标准液一定量,混匀,按“1.3”项下操作,测得回收率结果见表1.
, http://www.100md.com
Tab.1 Recovery of amoxicillin in urine Amount added/μg.mL-1
Amount found/μg.mL-1
Recovery/%
Average recovery/%
RSD/%
6.0
5.955
99.2
6.0
, http://www.100md.com
6.070
101.2
30.0
29.60
98.7
99.1±1.25
1.26
30.0
29.22
97.4
60.0
59.63
99.4
, 百拇医药
60.0
59.17
98.6
2.4 精密度试验
精密吸取羟氨苄青霉素标准液0.5,2.5,5.0 mL分置于50 mL量瓶中,依次加入1.0 mL空白滤过尿液,做日内、日间精密度试验,结果见表2.Tab.2 The precision determination of amoxicillin in urine Added/μg.mL-1
Within-day
Day to day
±s/μg.mL-1
, http://www.100md.com
RSD/%
±s/μg.mL-1
RSD/%
6.0
5.970±0.120
2.01
6.15±0.147
2.39
30.03
30.05±0.544
1.81
, 百拇医药
29.51±0.867
2.94
60.0
59.17±0.383
0.65
59.30±0.610
1.03
2.5 羟氨苄青霉素胶囊药动学参数的测定
精密吸取采集的过滤尿样1.0 mL,置50 mL量瓶中,按“1.3”项下操作,测得的数据应用实用药代动力学计算程序(3p87,中国药理学会数学药理专业委员会),由计算机自动计算,判断羟氨苄青霉素在人体内动态经时过程为单室模型,权重系数为1/C2,计算得出羟氨苄青霉素胶囊试验制剂与对照制剂药代动力参数见表3,4.Tab.3 Pharmacokinetic parameters of the test amoxicillin capsules No.
, http://www.100md.com
Ke/h-1
Ka/h-1
t1/2(Ka)/h
t1/2(Ke)/h
Tpeak/h
Cmax/μg.mL-1
AUC/μg.mL-1.h
1
, 百拇医药
0.453 7
1.289 4
0.537 6
1.527 8
1.249 8
1416.717 4
5 505.315 4
2
0.202 4
0.218 6
3.170 2
3.424 2
, http://www.100md.com
4.752 1
207.732 5
2 685.414 1
3
0.545 9
0.680 2
1.019 0
1.269 8
1.637 8
996.241 5
4 462.091 8
4
, 百拇医药
0.252 2
0.391 8
1.769 1
2.748 8
3.155 8
303.700 3
2 669.124 0
5
0.418 5
0.462 9
1.497 4
1.656 2
, 百拇医药
2.271 1
706.577 9
4 367.511 7
6
0.297 2
0.336 4
2.060 4
2.332 5
3.160 7
332.358 4
2 860.994 1
mean
, http://www.100md.com
0.361 6
0.563 2
1.675 6
2.159 9
2.704 6
660.554 6
3 758.408 4
s
0.13
0.39
0.91
0.83
, 百拇医药 1.27
474.70
1 188.37
Tab.4 Pharmacokinetic parameters of the control amoxicillin capsules No.
Ke/h-1
Ka/h-1
t1/2(Ka)/h
t1/2(Ke)/h
Tpeak/h
, http://www.100md.com Cmax/μg.mL-1
AUC/μg.mL-1.h
1
0.625 2
0.665 2
1.042 0
1.108 7
1.550 4
295.619 5
1 246.462 9
, 百拇医药
2
0.462 8
0.472 6
1.466 5
1.497 9
2.138 2
180.528 8
1 049.335 9
3
0.459 4
0.474 7
1.460 1
, 百拇医药
1.508 7
2.141 2
397.849 7
2 315.906 2
4
0.438 1
0.442 2
1.567 6
1.582 2
2.272 1
241.502 3
1 491.578 1
, 百拇医药
5
0.574 6
0.616 2
1.124 9
1.206 3
1.680 2
461.218 4
2 107.832 0
6
0.284 9
0.302 0
2.294 9
, http://www.100md.com
2.433 0
3.408 6
133.642 1
1 238.781 3
mean
0.474 2
0.495 5
1.492 7
1.556 1
2.198 4
285.060 2
1 574.982 8
, 百拇医药
s
0.19
0.13
0.44
0.66
0.47
126.19
517.13
参考文献
1,El Walily AM,El Sayed,M Abdel Hady,et al.Assay for amoxicillin in urine using derivative spectrophotometry. J Clin Pharm Ther,1992,17(2):101~105
, http://www.100md.com
2,孔晓东,蔡玉民.羟氨苄青霉素药物动力学研究.沈阳药学院学报,1986,3(4):241~244
3,Haginaka Jun, Wakai, Junko,et al. Liquid chromatographic determination of amoxicillin and its metabolites in human urine by postcolumn degradation with sodium hypochlorite. J Chromatogr, 1987,413:219~221
4,Carlgvist Jan, Westerlund Douglas.Automated determination of amoxicillin in biological fluids by column switching in ion-column derivatization. J Chromatogr,1985,344:285~286
5,Plavsic Fraujo, Habus Ivan. Chemical methods for determination of the concentration of some antihitics and fungicides in biological fluids. Farm Glas,1982,38(3):77~79
6,Nunez Vergara LJ,Squella JA,Silva MM.Polarographicassay for amoxicillin.Farmaco,1980,35(8):409~510
收稿日期:2000-03-19, 百拇医药
单位:王焕(鞍钢铁东医院药剂科,鞍山 114002);江晓君(武警辽宁总队医院药剂科,沈阳 110026);满羽(中国医科大学第一临床学院药剂科,沈阳 110001)
关键词:硫醇汞盐紫外分光光度法;羟氨苄青霉素;药代动力学
沈阳药科大学学报00zk08 摘 要 采用硫醇汞盐紫外分光光度法测定尿中羟氨苄青霉素的含量.方法回收率为(99.1±1.25)%,相对标准差为1.26%(n=6),线性范围6~60 μg/mL,日内、日间变异系数小于2.01%和2.94%,结果令人满意.
分类号 R914.1
A Study on the Determination of the Content of Amoxicillin in Urine
, http://www.100md.com
Wang Huan, Jiang Xiaojun, Man Yu
(The Tiedong Hospital of An′gang,Anshan 114002)
Abstract The determination of amoxicillin in urine by UV-spectrophotometry has been developed in this paper. The mean recovery and RSD were(99.1±1.25)% and 1.26%(n=6), respectively. The linearity of amoxicillin was from 6~60 μg.mL-1, and the validations of inter-day and intra-day were less than 2.01% and 2.94%, respectively.
, 百拇医药
Key words UV-spectrophotometry;Amoxicillin;pharmacokinetics
羟氨苄青霉素是广谱抗生素,广泛应用于临床.文献报道羟氨苄青霉素体内测定方法有紫外分光光度法〔1,2〕、高效液相色谱法〔3,4〕、荧光法〔5〕和极谱法〔6〕等.本研究采用硫醇汞盐紫外分光光度法对尿中羟氨苄青霉素的含量进行测定.测得本法平均回收率为(99.1±1.25)%,相对标准差为1.26%(n=6),日内、日间变异系数小于2.01%和2.94%.测定了6名男性健康自愿受试者交叉口服羟氨苄青霉素两种胶囊剂的羟氨苄青霉素药动学参数.
1 材料与方法
1.1 仪器与试剂
日本岛津-260型分光光度计(日本岛津),WFZ 800-D2型分光光度计(北京第二光学仪器厂),pH-25型酸度计,586型计算机.羟氨苄青霉素对照品(中国药品生物制品检定所提供),羟氨苄青霉素胶囊(试验制剂,批号:980809),羟氨苄青霉素胶囊(对照制剂,深圳华南制药有限公司提供,批号:990801).其它试剂均为分析纯.
, 百拇医药
1.2 供试液的配制
羟氨苄青霉素标准液(0.6 mg/mL):取羟氨苄青霉素对照品约0.15 g,精密称定,置250 mL量瓶中,加水溶解并稀释至刻度,摇匀,即得,置冰箱中保存.
硼酸缓冲液(pH 9.0):称取硼酸1.24 g,加水180 mL溶解后,加0.1 mol/L氢氧化钠溶液调pH至9.0,再用水稀释至200 mL,摇匀,即得.
醋酐乙腈溶液:取醋酐1 mL,加乙腈至50 mL,摇匀,即得.
咪唑溶液:取用苯精制过的咪唑8.25 g,加水60 mL溶解后,加盐酸(6 mol/L)8.3 mL,在搅拌下滴加0.27%二氯化汞溶液10 mL,调节pH值6.8±0.05,用水稀释至100 mL,滤过,摇匀,即得.
1.3 测定方法
, http://www.100md.com
精密吸取羟氨苄青霉素标准液或尿液适量,置50 mL量瓶中,加水稀释至刻度,摇匀,精密量取5.0 mL,置25 mL量瓶中,加硼酸缓冲液2.5 mL,醋酐乙腈溶液0.25 mL,放置5 min后,加咪唑溶液至刻度,摇匀,置60℃水浴中,加热30 min,取出、冷却.以水5.0 mL代替尿样,同法操作为空白,于325 nm处测定.
1.4 尿样的采集
男性健康自愿受试者6人,年龄22~24周岁,体重54~65 kg,随机分为两组,每组3人.试验前两周不得服用任何药物,前三天统一进标准餐、标准饮料,晨起空腹交叉服药(试验制剂含羟氨苄青霉素1.5 g,对照制剂含羟氨苄青霉素0.5 g),服药后2 h进标准餐,每次采样后饮温水100 mL,于服药后0.5,1.0,1.5,2.0,3.0,4.0,6.0,8.0 h采集尿液,量取体积后过滤,测定.
2 结果与讨论
, 百拇医药
2.1 测定条件的选择
精密吸取羟氨苄青霉素标准液两份2.5 mL,置50 mL量瓶中,其中一份加入空白滤过尿液1.0 mL,另取1.0 mL空白滤过尿液,按“1.3”项下操作,于250~400 nm间分别绘制吸收光谱,选取325 nm处测定时,空白尿液不干扰.
2.2 标准曲线的绘制
精密吸取羟氨苄青霉素标准液0.5,2.0,2.5,3.0,5.0 mL,分置50 mL量瓶中,按“1.3”项下操作,得回归方程为A=0.009 487+0.065 28C,r=0.999 2,线性范围为6~60 μg/mL,每隔30 min测定一次,结果表明:吸收度在2 h内不变.
2.3 回收率试验
取1.0 mL空白滤过尿液6份,分别精密加入羟氨苄青霉素标准液一定量,混匀,按“1.3”项下操作,测得回收率结果见表1.
, http://www.100md.com
Tab.1 Recovery of amoxicillin in urine Amount added/μg.mL-1
Amount found/μg.mL-1
Recovery/%
Average recovery/%
RSD/%
6.0
5.955
99.2
6.0
, http://www.100md.com
6.070
101.2
30.0
29.60
98.7
99.1±1.25
1.26
30.0
29.22
97.4
60.0
59.63
99.4
, 百拇医药
60.0
59.17
98.6
2.4 精密度试验
精密吸取羟氨苄青霉素标准液0.5,2.5,5.0 mL分置于50 mL量瓶中,依次加入1.0 mL空白滤过尿液,做日内、日间精密度试验,结果见表2.Tab.2 The precision determination of amoxicillin in urine Added/μg.mL-1
Within-day
Day to day
±s/μg.mL-1
, http://www.100md.com
RSD/%
±s/μg.mL-1
RSD/%
6.0
5.970±0.120
2.01
6.15±0.147
2.39
30.03
30.05±0.544
1.81
, 百拇医药
29.51±0.867
2.94
60.0
59.17±0.383
0.65
59.30±0.610
1.03
2.5 羟氨苄青霉素胶囊药动学参数的测定
精密吸取采集的过滤尿样1.0 mL,置50 mL量瓶中,按“1.3”项下操作,测得的数据应用实用药代动力学计算程序(3p87,中国药理学会数学药理专业委员会),由计算机自动计算,判断羟氨苄青霉素在人体内动态经时过程为单室模型,权重系数为1/C2,计算得出羟氨苄青霉素胶囊试验制剂与对照制剂药代动力参数见表3,4.Tab.3 Pharmacokinetic parameters of the test amoxicillin capsules No.
, http://www.100md.com
Ke/h-1
Ka/h-1
t1/2(Ka)/h
t1/2(Ke)/h
Tpeak/h
Cmax/μg.mL-1
AUC/μg.mL-1.h
1
, 百拇医药
0.453 7
1.289 4
0.537 6
1.527 8
1.249 8
1416.717 4
5 505.315 4
2
0.202 4
0.218 6
3.170 2
3.424 2
, http://www.100md.com
4.752 1
207.732 5
2 685.414 1
3
0.545 9
0.680 2
1.019 0
1.269 8
1.637 8
996.241 5
4 462.091 8
4
, 百拇医药
0.252 2
0.391 8
1.769 1
2.748 8
3.155 8
303.700 3
2 669.124 0
5
0.418 5
0.462 9
1.497 4
1.656 2
, 百拇医药
2.271 1
706.577 9
4 367.511 7
6
0.297 2
0.336 4
2.060 4
2.332 5
3.160 7
332.358 4
2 860.994 1
mean
, http://www.100md.com
0.361 6
0.563 2
1.675 6
2.159 9
2.704 6
660.554 6
3 758.408 4
s
0.13
0.39
0.91
0.83
, 百拇医药 1.27
474.70
1 188.37
Tab.4 Pharmacokinetic parameters of the control amoxicillin capsules No.
Ke/h-1
Ka/h-1
t1/2(Ka)/h
t1/2(Ke)/h
Tpeak/h
, http://www.100md.com Cmax/μg.mL-1
AUC/μg.mL-1.h
1
0.625 2
0.665 2
1.042 0
1.108 7
1.550 4
295.619 5
1 246.462 9
, 百拇医药
2
0.462 8
0.472 6
1.466 5
1.497 9
2.138 2
180.528 8
1 049.335 9
3
0.459 4
0.474 7
1.460 1
, 百拇医药
1.508 7
2.141 2
397.849 7
2 315.906 2
4
0.438 1
0.442 2
1.567 6
1.582 2
2.272 1
241.502 3
1 491.578 1
, 百拇医药
5
0.574 6
0.616 2
1.124 9
1.206 3
1.680 2
461.218 4
2 107.832 0
6
0.284 9
0.302 0
2.294 9
, http://www.100md.com
2.433 0
3.408 6
133.642 1
1 238.781 3
mean
0.474 2
0.495 5
1.492 7
1.556 1
2.198 4
285.060 2
1 574.982 8
, 百拇医药
s
0.19
0.13
0.44
0.66
0.47
126.19
517.13
参考文献
1,El Walily AM,El Sayed,M Abdel Hady,et al.Assay for amoxicillin in urine using derivative spectrophotometry. J Clin Pharm Ther,1992,17(2):101~105
, http://www.100md.com
2,孔晓东,蔡玉民.羟氨苄青霉素药物动力学研究.沈阳药学院学报,1986,3(4):241~244
3,Haginaka Jun, Wakai, Junko,et al. Liquid chromatographic determination of amoxicillin and its metabolites in human urine by postcolumn degradation with sodium hypochlorite. J Chromatogr, 1987,413:219~221
4,Carlgvist Jan, Westerlund Douglas.Automated determination of amoxicillin in biological fluids by column switching in ion-column derivatization. J Chromatogr,1985,344:285~286
5,Plavsic Fraujo, Habus Ivan. Chemical methods for determination of the concentration of some antihitics and fungicides in biological fluids. Farm Glas,1982,38(3):77~79
6,Nunez Vergara LJ,Squella JA,Silva MM.Polarographicassay for amoxicillin.Farmaco,1980,35(8):409~510
收稿日期:2000-03-19, 百拇医药