CN104880442A - 一种测定盐酸多巴胺的方法 - Google Patents
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- CTENFNNZBMHDDG-UHFFFAOYSA-N Dopamine hydrochloride Chemical compound Cl.NCCC1=CC=C(O)C(O)=C1 CTENFNNZBMHDDG-UHFFFAOYSA-N 0.000 title claims abstract description 38
- 229960001149 dopamine hydrochloride Drugs 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000000243 solution Substances 0.000 claims abstract description 19
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 230000005284 excitation Effects 0.000 claims abstract description 6
- 239000007853 buffer solution Substances 0.000 claims abstract description 4
- GKAMNGMEOQWSHF-UHFFFAOYSA-L potassium;sodium;chloride;hydroxide Chemical compound [OH-].[Na+].[Cl-].[K+] GKAMNGMEOQWSHF-UHFFFAOYSA-L 0.000 claims abstract description 4
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 claims description 16
- 229960003638 dopamine Drugs 0.000 claims description 8
- 238000012417 linear regression Methods 0.000 claims description 3
- GVKCHTBDSMQENH-UHFFFAOYSA-L phloxine B Chemical compound [Na+].[Na+].[O-]C(=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C([O-])=C(Br)C=C21 GVKCHTBDSMQENH-UHFFFAOYSA-L 0.000 claims 1
- 238000002347 injection Methods 0.000 abstract description 6
- 239000007924 injection Substances 0.000 abstract description 6
- 239000012153 distilled water Substances 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 241000220317 Rosa Species 0.000 abstract description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000001514 detection method Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940090044 injection Drugs 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
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- 208000018737 Parkinson disease Diseases 0.000 description 1
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- 206010040047 Sepsis Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000007675 cardiac surgery Methods 0.000 description 1
- 150000003943 catecholamines Chemical class 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940005482 dopamine injection Drugs 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 230000008482 dysregulation Effects 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000000695 excitation spectrum Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
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- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
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Abstract
本发明公开了一种测定盐酸多巴胺的方法。于7支10mL比色管中分别加入0.00、0.05、0.1、0.2、0.5、0.9、1.3mL 7.0×10- 4~9.0×10- 4mol/L的盐酸多巴胺溶液,再在每支管中加入0.5~3.0mL pH12.0~13.0的氯化钾-氢氧化钠缓冲液和0.2~0.9mL 1.0×10- 5mol/L的荧光桃红溶液;以不加盐酸多巴胺的溶液为试剂空白,用二次蒸馏水定容至刻度,静置20分钟后,于荧光光度计上,以536nm为激发波长,551nm为发射波长,用1cm荧光比色皿,分别测定含盐酸多巴胺溶液的荧光值F和试剂空白的荧光值F0,计算差值ΔF= F0-F;另取盐酸多巴胺注射液1支转入100mL容量瓶中并定容,同法测定荧光值,计算出注射液中盐酸多巴胺的含量。本发明灵敏度高、线性范围宽、操作简便。
Description
技术领域
本发明涉及盐酸多巴胺的测定方法,特别是荧光桃红荧光猝灭法测定盐酸多巴胺的方法。
背景技术
多巴胺(Dopamine, DA)是一种最重要的儿茶酚胺,在中枢神经系统及肾脏系统中起着关键的作用。体内的多巴胺分泌失调将导致心脏病、精神分裂症和帕金森氏症等疾病。盐酸多巴胺水溶性好、可增强心肌收缩力,增加排血量,并大量用于心肌梗塞、创伤、内毒素败血症、心脏手术、肾功能衰竭、充血性心力衰竭等引起的休克综合征。因此,建立一种灵敏、快速、简单的多巴胺检测方法,对生物分析、生理学、临床医学以及相关药物的质量控制研究具有十分重要的意义。目前,测定多巴胺的方法有电化学法、色谱法、化学发光法等。而荧光法具有分析灵敏度高、方法简便、仪器价廉等优点,常应用于药物分析检测。荧光桃红(RA)是一种性能优异的强荧光物质,常作为荧光试剂用于定量分析,但目前应用荧光桃红测定盐酸多巴胺的方法尚未见报道。
发明内容
本发明的目的是提供一种荧光桃红荧光猝灭法测定盐酸多巴胺的方法。
具体步骤为:
在7支10mL比色管中分别加入0.00、0.05、0.1、0.2、0.5、0.9、1.3mL 7.0×10-4~9.0×10-4mol/L的盐酸多巴胺(DA)溶液,再在每支比色管中都分别加入0.5~3.0mL pH12.0~13.0的氯化钾-氢氧化钠缓冲溶液和0.2~0.9mL 1.0×10-5mol/L的荧光桃红(RA)溶液;以不加盐酸多巴胺的溶液为试剂空白,二次蒸馏水定容至刻度,摇匀,静置20分钟后,于荧光光度计上,设置激发波长为536nm,发射波长为551nm,激发和发射狭缝均为2.5nm,电压为600V,用1cm荧光比色皿,分别测定含盐酸多巴胺溶液的荧光强度值F和试剂空白的荧光强度值F0,计算荧光强度差值ΔF= F0-F; 其荧光强度差值ΔF与盐酸多巴胺浓度C在4.03~105μmol/L范围内成线性关系,线性回归方程为:ΔF=2.532C+19.04(C为盐酸多巴胺的浓度,单位为μmol/L), 相关系数0.9992,检出限为1.26μmol/L;另取盐酸多巴胺注射液1支,全部转移至100mL容量瓶中,用二次蒸馏水定容至刻度,取0.1~0.3mL溶液同法测定荧光强度值,计算出盐酸多巴胺注射液中盐酸多巴胺的含量。
本发明测定方法灵敏度高、线性范围宽、操作简便。
附图说明
图1为本发明实施例空白与1.05×10-4 mol/L盐酸多巴胺的荧光光谱图。
图中标记a:pH13.0 KCl-NaOH – 0.7μmol/L RA的激发光谱;b:pH13.0 KCl-NaOH – 0.7μmol/L RA的发射光谱;c:pH13.0 KCl-NaOH – 0.7μmol/L RA-1.05×10-4mol/L DA的发射光谱。
具体实施方式
实施例:
在7支10mL比色管中分别加入0.00、0.05、0.1、0.2、0.5、0.9、1.3mL 8.068×10-4mol/L的盐酸多巴胺溶液,再在每支比色管中都分别加入2.5mL pH13.0的氯化钾-氢氧化钠缓冲溶液和0.7mL 1.0×10-5mol/L的荧光桃红溶液,以不加盐酸多巴胺的溶液为试剂空白,二次蒸馏水定容至刻度,摇匀,静置20分钟。于荧光分光光度计上(Cary Eclipse型,美国Varian公司),设置激发波长为536nm,发射波长为551nm,激发和发射狭缝均为2.5nm,电压为600V,用1cm荧光比色皿,分别测定含盐酸多巴胺溶液的荧光强度值F和试剂空白的荧光强度值F0,计算荧光强度差值ΔF= F0-F; 其荧光强度差值ΔF与盐酸多巴胺浓度C在4.03~105μmol/L范围内成线性关系,线性回归方程为:ΔF=2.532C+19.04(C为盐酸多巴胺的浓度,单位为μmol/L), 相关系数0.9992,检出限为1.26μmol/L。另取两个厂家(广东白云山明兴制药有限公司和上海禾丰制药厂,规格:盐酸多巴胺含量20mg/2mL)生产的盐酸多巴胺注射液各一支,分别全部转移至100mL容量瓶中,用二次蒸馏水定容至刻度,各取0.2mL溶液同法测定荧光强度值,计算出盐酸多巴胺注射液中盐酸多巴胺的含量,同时做了加标回收实验,结果见表1。
表1:盐酸多巴胺注射液测定结果(n=5)
样品 | 测定值(mg/2mL) | RSD(%) | 标示量(mg/2mL) | 回收率( %) |
1 | 20.06 | 1.35 | 20 | 108.15 |
2 | 19.91 | 1.36 | 20 | 108.56 |
Claims (1)
1.一种测定盐酸多巴胺的方法,其特征在于具体步骤为:
在7支10mL比色管中分别加入0.00、0.05、0.1、0.2、0.5、0.9、1.3mL 7.0×10-4~9.0×10-4mol/L的盐酸多巴胺溶液,再在每支比色管中都分别加入0.5~3.0mL pH12.0~13.0的氯化钾-氢氧化钠缓冲溶液和0.2~0.9mL 1.0×10-5mol/L的荧光桃红溶液;以不加盐酸多巴胺的溶液为试剂空白,二次蒸馏水定容至刻度,摇匀,静置20分钟后,于荧光光度计上,设置激发波长为536nm,发射波长为551nm,激发和发射狭缝均为2.5nm,电压为600V,用1cm荧光比色皿,分别测定含盐酸多巴胺溶液的荧光强度值F和试剂空白的荧光强度值F0,计算荧光强度差值ΔF= F0-F; 其荧光强度差值ΔF与盐酸多巴胺浓度C在4.03~105μmol/L范围内成线性关系,线性回归方程为:ΔF=2.532C+19.04,C为盐酸多巴胺的浓度,单位为μmol/L, 相关系数0.9992,检出限为1.26μmol/L;另取盐酸多巴胺注射液1支,全部转移至100mL容量瓶中,用二次蒸馏水定容至刻度,取0.1~0.3mL溶液同法测定荧光强度值,计算出盐酸多巴胺注射液中盐酸多巴胺的含量。
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CN112945878A (zh) * | 2021-02-03 | 2021-06-11 | 安阳市妇幼保健院(安阳市儿童医院) | 间接光度法测定多巴胺的方法 |
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CN103980716A (zh) * | 2014-04-11 | 2014-08-13 | 中国科学院化学研究所 | 一种测定多巴胺含量的方法 |
CN104181135A (zh) * | 2014-08-19 | 2014-12-03 | 东南大学 | 水溶性硅量子点在多巴胺检测中的应用 |
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Patent Citations (4)
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EP2055384A1 (de) * | 2007-10-31 | 2009-05-06 | Leukocare AG | Vorrichtung zum Nachweis von Bestandteilen in einem Fluid |
CN102288763A (zh) * | 2011-07-06 | 2011-12-21 | 清华大学深圳研究生院 | 一种基于量子点的免疫荧光检测莱克多巴胺的方法及专用试剂盒 |
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CN112945878A (zh) * | 2021-02-03 | 2021-06-11 | 安阳市妇幼保健院(安阳市儿童医院) | 间接光度法测定多巴胺的方法 |
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