CN103760149A - Method for detecting propranlolum rapidly and flexibly based on nanogold chemiluminiscence - Google Patents
Method for detecting propranlolum rapidly and flexibly based on nanogold chemiluminiscence Download PDFInfo
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- CN103760149A CN103760149A CN201410015916.4A CN201410015916A CN103760149A CN 103760149 A CN103760149 A CN 103760149A CN 201410015916 A CN201410015916 A CN 201410015916A CN 103760149 A CN103760149 A CN 103760149A
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Abstract
The invention discloses a method for detecting propranlolum rapidly and flexibly based on nanogold chemiluminiscence. The method comprises the following steps: 1) preparing a nanogold solution by adopting a sodium borohydride reduction method; 2) analyzing and determining the propranlolum by adopting a flow injection chemiluminiscence analysis system, wherein the particular flow path is as follows: firstly, mixing a luminal solution and H2O2, introducing the nanogold and propranlolum solution into an analysis system by taking secondary deionized water as carrying flow, introducing the luminal solution, the H2O2 and the secondary deionized water into a circulation tank by a peristaltic pump, injecting the mixed solution of nanogold and the propranlolum by a sample injection valve; detecting chemiluminiscence signals by a photomultiplier close to a flow injection chemiluminiscence tank; and then inputting the chemiluminiscence signals into a computer, and recording and collecting data. The detection limit can be up to 8.2*10<-12>g/mL, and the sensitivity is improved by one order compared with that of a conventional chemiluminiscence analysis method, and the analysis and testing time is shorted within 10 minutes.
Description
Technical field
The present invention relates to drug test technical field, specifically, the present invention relates to a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive.
Background technology
Inderal is a kind of beta-2 adrenoceptor retarding agent, and chemical name is 1-isopropylamino-3-(1-naphthoxy)-2-propylate hydrochlorate, is widely used in the treatment of angiocardiopathy, belongs to the medicine of anti-hypertension class.This medicine is used as excitant to improve sportsman's achievement in play in recent years, and the International Olympic Committee is classified as illegal drug.Therefore, its sensitive mensuration is seemed to very important.In addition, medicine sampling observation is a kind of important means of drug surveilance, and annual country will spend a large amount of funds and the time is carried out examination at random work.China basic unit drug surveilance department amount is large at present, and staff is few, mostly lacks technical equipment and the information resources of target examination at random.And the existing many report of the assay method of inderal character, as the mensuration of its solubleness, metastable state peak width, racemic character and by the gathering character of mass spectroscopy weak self-association system of inderal in water etc.But the method report of measuring quantitatively inderal is little, only has spectrophotometric method, lotus to move reaction method and chemiluminescence method and fluorometry etc.When fluorometry is measured the material demand of several fluorescence spectrum overlappings simultaneously, usually helpless.Pulgarin etc. utilize non-linear varied angle synchronous fluorimetry to solve this problem, but this method is comparatively harsh to the selection of experiment condition, and the searching of its angle and the selection of optimum point all need extremely rich experience.And other analytical approach such as spectrophotometric method all exists defects such as sensitivity is low, troublesome poeration.Therefore, do not increasing under the prerequisite of economy and time cost, studying and apply new method and solve this class problem and there is value.
Chemiluminescence analysis is highly sensitive because having, the range of linearity is wide, reagent is cheap and instrumentation is simple etc., and advantage has been widely used in Pharmaceutical Analysis.In recent years, nano particle especially noble metal nano particles has many advantages, and first, in nano-scale range, various nano particle with special performance is easy to synthesize; Secondly, nano particle has good biocompatibility, can not affect biologically active.Therefore, metal nanoparticle especially noble metal nano particles as biomarker, caused the very large concern of researcher.In recent years, noble metal nano particles is served as a mark thing was measured some medicines for chemiluminescence analysis and was more and more expanded.Haghighi in 2010 etc. utilize the chemical luminous system of Nano Silver catalysis, adopt the mode of flow injection to realize the mensuration of Antituberculous Drugs-isoniazid.Liu in 2012 etc. utilize the chemical luminous system of Nano Silver catalysis to realize the analysis of itrofurans medicine in food and measure.Du etc. utilizes the chemical luminous system of nano gold catalysis to measure the medicine timolol maleate that is used for the treatment of miocardial infarction disease.Rezaei etc. utilize butylphenol Catalyzed by Nano-ferric Oxide luminol-periodic acid silver potassium chemiluminescence reaction to measure aminophylline.Lu etc. utilize triangular nano gold catalysis luminol chemiluminescence system of determination captopril.Du in 2013 etc. utilize nano gold catalysis chemical analysis to measure Trimetazidine.Amjadi finds that captopril can suppress potassium permanganate-bromide chemiluminescence reaction of Nano Silver catalysis, has realized the analysis of captopril medicine is measured.The discovery Platinum Nanoparticles such as Lee can cause luminol-silver nitrate chemiluminescence reaction, realized micro-fluidic chemiluminescence analysis measure Cobastab based on this.In addition, there is report gold and silver core-shell nano to react by catalysis luminol chemiluminescence, thereby realize, the analysis of cancer therapy drug Drogenil is measured.
Before us, nm of gold because affecting it, has been carried out deeply and systematic research chemiluminescence reaction catalytic effect in the change of this form of reuniting.The reunion of nm of gold can be directly used in to chemiluminescence analysis and measure the reunion that inderal key is that can inderal induced nano gold, thereby further affect its cataluminescence performance.Have bibliographical information inderal the life of induced nano golden hair to reunite, our research simultaneously has also confirmed this result, and is applied to chemiluminescence analysis mensuration inderal.The analytical approach simple and sensitive based on nm of gold chemiluminescence analysis mensuration inderal that the present invention sets up, fast.Whole analysis is measured and will within 10 minutes, be completed.
Summary of the invention
The invention solves the limitation that conventional medicament assay method exists: use the instrument of large-scale costliness, analysis cost is high; Sensitivity can not meet the demand of each side; Troublesome poeration and having relatively high expectations to analyst; Analyze minute longer.Utilize the reunion that inderal can induced nano gold, thereby affect its catalytic activity to luminol chemiluminescence, set up a kind of method of brand-new simple and sensitive chemical luminescent detecting inderal.
The present invention is directed to the deficiency that current inderal detection method exists, utilize inderal can cause the reunion of nm of gold, thereby cause its change to luminol chemiluminescence catalytic performance, set up a kind of new method of simple, sensitive rapid chemical luminescence assays inderal.Measuring principle: when inderal does not exist, nano gold catalysis luminol-hydrogen peroxide system of dispersed, produces weak chemiluminescence signal; And when inderal exists, inderal causes nm of gold to be reunited, the nm of gold of reunion state strengthens the chemiluminescence signal of luminol-hydrogen peroxide system widely, thereby realizes the mensuration to inderal.
Technical scheme provided by the invention is: a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive, comprises the following steps:
1) nano-Au solution preparation, adopts sodium borohydride reduction to prepare nano-Au solution, is specially: under room temperature condition, by the HAuCl of 100mL0.01%w/w
41% citric acid three sodium solution of aqueous solution, 1mL and the 0.075%NaBH of 1mL
4/ 1% citric acid three sodium solution mixes completely, and after mixing, solution strong stirring 30 minutes, after complete reaction, is put in 4 ℃ of Refrigerator stores stand-by; The particle diameter of the nm of gold of selecting is 2.6nm, and concentration is 6.0 × 10
-7mol/L
2) adopt Flow Injection Analysis/Chemiluminescence systematic analysis to measure inderal, Flow Injection Analysis/Chemiluminescence system comprises an IFFM-D flow injection system, an IFFS-A luminescence detector and a control computer, and concrete stream is: luminol solution and H
2o
2after first mixing, secondary deionized water is brought nm of gold and inderal solution in analysis system into as current-carrying, luminol solution, H
2o
2with secondary deionized water by the peristaltic pump pond that circulates, the mixed solution of nm of gold and inderal injects by sampling valve, chemiluminescence signal detects by the photomultiplier in contiguous portable injection chemiluminescence pond, then inputs computer and carries out record and the collection of data.
Described step 2) in, selecting the concentration of luminol is 5 × 10
-4mol/L, concentration of hydrogen peroxide is 5 × 10
-2mol/L.
Described step 2) in, luminol solution, H
2o
2with secondary deionized water by the circulate flow velocity in pond of peristaltic pump, be 2.6mL/min.
Described step 2) in, the concentration range of inderal is 2.1 × 10
-9～6.0 × 10
-7g/mL.
Described step 2) in, the pH value of chemical luminous system is optimized for 12.0.
The invention has the beneficial effects as follows:
The present invention has the following advantages: (1) detection limit reaches 8.2 × 10
– 12g/mL, the sensitivity of chemiluminescence analysis method than before improves 1 order of magnitude; (2) the analytical test time shorten of the method is in 10 minutes; (3) condition of whole operation experiments is gentleer, and completes in the system of a homogeneous phase, is conducive to the analysis operation of robotization.
Accompanying drawing explanation
When considered in conjunction with the accompanying drawings, by the detailed description with reference to below, can more completely understand better the present invention and easily learn wherein many advantages of following, but accompanying drawing described herein is used to provide a further understanding of the present invention, form a part of the present invention, schematic description and description of the present invention is used for explaining the present invention, does not form inappropriate limitation of the present invention, wherein:
The flow circuit diagram of Fig. 1 Flow Injection Analysis/Chemiluminescence test;
The optimization of Fig. 2 nm of gold concentration;
The optimization of Fig. 3 luminol concentration;
The optimization of Fig. 4 concentration of hydrogen peroxide;
The selection of Fig. 5 system pH;
The sensitivity analysis of Fig. 6 chemiluminescence detection inderal;
The uv-vis spectra of nm of gold before and after Fig. 7 inderal adds, figure a is nm of gold; Figure b is nm of gold+inderal;
The transmission electron microscope picture of nm of gold before and after Fig. 8 inderal adds, figure a is before adding; Figure b is after adding.
Embodiment
For above-mentioned purpose of the present invention, feature and advantage can be become apparent more, below in conjunction with the drawings and specific embodiments, the present invention is further detailed explanation.
Key instrument equipment used and reagent:
IFFL-D Flow Injection Analysis/Chemiluminescence instrument and support equipment thereof (Xi'an Rui Mai Analytical Instrument Co., Ltd), TU-1901 type spectrophotometry instrument (Beijing Pu Xi all purpose instrument company limited), transmission electron microscope (Hitachi, Ltd, specifications and models: H-600).
Gold chloride (HAuCl
44H
2o, AR, Au content >47.8%, Chemical Reagent Co., Ltd., Sinopharm Group); Gold chloride solid is dissolved in secondary deionized water and prepares gold chloride stock solution, be put in 4 ℃ of refrigerators and preserve.4.43g luminol pressed powder is dissolved in 20mL0.10M NaOH solution and is diluted to 1L, prepare 2.5 × 10
2m luminol stock solution.Before using, keep in Dark Place a week to guarantee the stable of reagent character.The working solution of luminol obtains by the dilution of stock solution.H
2o
2working solution when each use from 30% (w/w) H
2o
2(Solution on Chemical Reagents in Shanghai company) dilution obtains.Inderal is purchased from Chinese medicine and biological products management office, and experimental water is secondary deionized water.
The optimization of experiment condition:
In experiment, select the nm of gold of 2.6nm, examined or check the impact of golden concentration.As Fig. 2, the concentration of gold is larger, and the luminous intensity of system is also larger, and it is 2.6nm that the present invention selects the particle diameter of nm of gold, and concentration is 6.0 × 10
-7mol/L.
Investigated the impact of chemical illuminating reagent, as Fig. 3 and 4, found the increase along with luminol and concentration of hydrogen peroxide, the luminous intensity of system is larger.Consider the consumption of chemiluminescence intensity and reagent, selecting the concentration of luminol is 5 × 10
-4mol/L, concentration of hydrogen peroxide is 5 × 10
-2mol/L.
Investigated the impact of system pH, as Fig. 5, when the pH of system value is less than 12, along with the increase chemiluminescence intensity of pH value increases; And when the pH of system value is greater than 12, along with the increase chemiluminescence intensity of pH value weakens.The pH value of the present invention's system used is optimized for 12.0.
The preparation of 2.6nm nm of gold:
In preparation process, glassware used is all used chloroazotic acid to clean (1:3 HNO
3– HCl), then use secondary deionized water cleaning down clean, dry for standby.The preparation method of 2.6nm nm of gold is sodium borohydride reduction.Preparation process is summarized as follows: under the condition of room temperature, strong stirring, by the HAuCl of 100mL
4(0.01%w/w) 1% citric acid three sodium solution of aqueous solution, 1mL and the 0.075%NaBH of 1mL
4/ 1% citric acid three sodium solution mixes completely.After mixing, solution stirs 30 minutes, after complete reaction, is put in 4 ℃ of Refrigerator stores stand-by.
Based on nm of gold Flow Injection Analysis/Chemiluminescence, measure inderal:
Flow Injection Analysis/Chemiluminescence system comprises an IFFM-D flow injection system, an IFFS-A luminescence detector and a control computer.Adopt stream as shown in Figure 1, under current-carrying intermediate water carries, nm of gold and luminol and H
2o
2mixed solution mix.The present invention is by luminol and H
2o
2after first mixing, secondary deionized water is brought nano-Au solution in analysis system into as current-carrying, luminol solution, H
2o
2by the peristaltic pump pond that circulates, flow velocity is 2.6mL/min with secondary deionized water.The mixed solution (nano-Au solution of reunion) of nm of gold and nm of gold and inderal injects by sampling valve, and volume is about 150 μ L.Chemiluminescence signal detects by the photomultiplier (PMT) in contiguous portable injection chemiluminescence pond, then inputs computer and carries out record and the collection of data.
The chemiluminescent analytical performance of inderal:
Under the experiment condition of above-mentioned optimization, realized the mensuration to inderal chemiluminescence analysis.Result shows, along with the increase of inderal concentration, chemiluminescence signal strengthens gradually.And 2.1 × 10
-9～6.0 × 10
-7in the concentration range of g/mL, the concentration of inderal becomes good correlativity with the intensity of chemiluminescence signal, and as shown in Figure 6, it detects and is limited to 8.2 × 10 linearity curve
12g/mL (S/N=3).And to 6.2 × 10
8the inderal of g/mL carries out replication five times, and relative standard deviation (R.S.D.) is 4.6%.Experimental result shows that the method detection limit is low, and precision is good.Compare with the method that inderal is measured in only a kind of chemiluminescence analysis, the method detection limit that the present invention sets up will a low order of magnitude.
The result of sample analysis:
Get 10 of commercially available propranolol hydrochloride tablets agent, be placed in carefully porphyrize of mortar.Powdered sample is added in small beaker to a small amount of water-soluble solution, then constant volume in 100mL volumetric flask, standing filtration.Above-mentioned solution carries out luminesceence analysis after dilution.Replicate determination 5 times, and by pharmacopeia method (ultraviolet spectrophotometry), Propranolol Hydrochloride sample is carried out to check analysis, analysis result is listed in table 1.Result shows that two kinds of methods record result and there is no obvious difference.
The result of table 1 sample analysis.
Utilize two kinds of means of uv-visible absorption spectra and transmission electron microscope to characterize the mixed solution of 2.6nm nm of gold and 2.6nm nm of gold and inderal, result is as Fig. 7 and Fig. 8.As shown in Figure 7, there is obvious change in ultraviolet-visible absorption spectroscopy.In addition, from the result (Fig. 8) of transmission electron microscope, can find out, before inderal adds, nm of gold is that monodispersed (a), (Fig. 8 b) to add rear nm of gold that obvious reunion has occurred for Fig. 8.
The explanation of above example is just for helping to understand core concept of the present invention; , for one of ordinary skill in the art, according to thought of the present invention, all will change in specific embodiments and applications, in sum, this description should not be construed as limitation of the present invention meanwhile.
Claims (6)
1. a method that detects inderal based on nm of gold chemiluminescence rapid sensitive, is characterized in that, comprises the following steps:
1) nano-Au solution preparation, adopts sodium borohydride reduction to prepare nano-Au solution, is specially: under room temperature condition, by the HAuCl of 100mL0.01%w/w
41% citric acid three sodium solution of aqueous solution, 1mL and the 0.075%NaBH of 1mL
4/ 1% citric acid three sodium solution mixes completely, and after mixing, solution strong stirring 30 minutes, after complete reaction, is put in 4 ℃ of Refrigerator stores stand-by;
2) adopt Flow Injection Analysis/Chemiluminescence systematic analysis to measure inderal, Flow Injection Analysis/Chemiluminescence system comprises an IFFM-D flow injection system, an IFFS-A luminescence detector and a control computer, and concrete stream is: luminol solution and H
2o
2after first mixing, secondary deionized water is brought nm of gold and inderal solution in analysis system into as current-carrying, luminol solution, H
2o
2with secondary deionized water by the peristaltic pump pond that circulates, the mixed solution of nm of gold and inderal injects by sampling valve, chemiluminescence signal detects by the photomultiplier in contiguous portable injection chemiluminescence pond, then inputs computer and carries out record and the collection of data.
2. a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive according to claim 1, is characterized in that, in described step 1), the particle diameter of the nm of gold of selecting is 2.6nm, and concentration is 6.0 × 10
-7mol/L.
3. a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive according to claim 1, is characterized in that described step 2) in, selecting the concentration of luminol is 5 × 10
-4mol/L, concentration of hydrogen peroxide is 5 × 10
-2mol/L.
4. a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive according to claim 1, is characterized in that described step 2) in, luminol solution, H
2o
2with secondary deionized water by the circulate flow velocity in pond of peristaltic pump, be 2.6mL/min.
5. a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive according to claim 1, is characterized in that described step 2) in, the concentration range of inderal is 2.1 × 10
-9～6.0 × 10
-7g/mL.
6. a kind of method that detects inderal based on nm of gold chemiluminescence rapid sensitive according to claim 1, is characterized in that described step 2) in, the pH value of chemical luminous system is optimized for 12.0.
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| CN108802013A (en) * | 2018-05-25 | 2018-11-13 | 江南大学 | A method of the chemoluminescence method based on nano platinum particle detects mercury ion |
| CN115128062A (en) * | 2022-08-29 | 2022-09-30 | 中储粮成都储藏研究院有限公司 | Method for detecting freshness of grains and application |
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