| Objective: The organism will respond positively to the stressor in daily life when stressor act on it. If the stressor is not strong enough, the response of body to stressor will be beneficial to the body ,which inspire the body potentiality and to promote the body to complete the hard work or avoid the possible risk.Oppositely when the stressor is too intense or long-lasting,it will lead to organism metabolic disorder and damage,which is harmful to organism.The trauma as a intense stressor often lead to diseases such as the anabrosis of stomach, superficial ulcer, stress ardiac dysrhythmia, heart disease, myocardial infarction.The occurrence and development of these disease is relative to the catecholamine when organism is stressing.If we set up a simple,rapid detection tool for the concentration of catecholamines in the blood and find the diseases dependability on CA,then we can interfere with the stress disease by intervention messures which hamper the disease and protect the advantageous stress reaction. Flow-injection chemiluminescence detection as a new means is more and more widely used in medical field,which has the advantage such as simple equipment,high sensitivity,wide testing range,better selectivity, more easy controlling,etc. However, we set up a simple, rapid detection methods for the concentration of dopamine and epinephrine.Motheds: There are three CA: dopamine, epinephrine, norepinephrine.We have established methods to test dopamine, epinephrine. which is devid to two parts:Part one:The stock solution of Epinephrine :Epinephrine was dissolved in water to make a stock solution at a concentration of 2×10-4 mol L-1. The solution was stored at 4?C and diluted to working solutions with pure water. luminol solution: A 0.02 mol L-1 luminol solution was prepared by dissolving 0.886 g luminol in 7mL 1 mol L-1 NaOH and diluting with deionized water to 250 mL. Bis(hydrogen periodato)argentate(III) complex anion, [Ag(HIO6)2]5-, was synthesized according to the procedure described previously. Flow lines of The FI-CL System were inserted into the carrier flow, luminol, Ag(III) and sample solution, respectively. The pumps were started and the mixed solution flowed into the cell. The CL reaction occurred and was recorded. When the baseline was stable, the epinephrine solution was injected into the carrier stream, then merged with the reagent streams (Ag(III) and luminol mixture solution). The CL intensity vs. epinephrine concentration were used for the calibration. Calibration graphs were constructed by plotting the intensity (peak height minus base signal) of the CL signal against the concentration of epinephrine. Under the optimal conditions used for CL measurements,the FI with Ag(III)-luminol CL detection is used to determine the determination of adrenailne hydrochloride injection sample.Part two:The stock solution of dopamine:Dopamine hydrochloride was dissolved in distilled water to make a stock solution at a concentration of 0.1mg mL-1 (5.27×10-4 mol /L). The solution was stored at 4?C and diluted to working solutions with pure water step by step. A 0.02 mol /L luminol solution was prepared by dissolving 0.8860 g luminol in 7mL 1 mol L-1 NaOH and diluting with deionized water to 250 mL. Bis(hydrogen perioDopto)argentate(III) complex anion, [Ag(HIO6)2]5-, was synthesized according to the procedure described previously. Flow lines of The FI-CL System were inserted into the carrier flow, luminol, Ag(III) and sample solution, respectively. The pumps were started and the mixed solution flowed into the cell. The CL reaction occurred and was recorded. When the baseline was stable, the dopamine solution was injected into the carrier stream, then merged with the reagent streams (Ag(III) and luminol mixture solution). The CL intensity vs. dopamine concentration were used for the calibration. Calibration graphs were constructed by plotting the intensity (peak height minus base signal) of the CL signal against the concentration of dopamine. Under the optimal conditions used for CL measurements , the FI with Ag(III)-luminol CL detection is used to determine the determination of dopamine hydrochloride injection sample.Results:We use Flow-injection analysis with the Ag(III) Complex- luminol chemiluminescent system to test the epinephrine. Under the optimum conditions( the flow rate of 2.5ml/min, deionized water as carrier flow, concentration of luminol choose as 2.0×10-7 mol/L , alkaline concentration of luminol choose as 0.005 mol/L , concentration of Ag(III) chosen as 1.0×10-5 mol /L and alkaline concentration of Ag(III) was 0.040mol/L),The detection limit (3σ) was 8.0×10-10 mol /L, plots of the CL intensity versus concentration of epinephrine were linear in the range of 1.0×10-9 ? 1.0×10?7 mol /L.The regression equation was I = -88.4 + 1843.6×108 C (r = 0.9991). The RSD (n = 11) was 2.9 % for 1.5×10-8 mol/L epinephrine. We use Flow-injection analysis with the Ag(III) Complex-luminol chemiluminescent system to test the dopamine. Under the optimum conditions( the flow rate of 2.5ml/min, deionized water as carrier flow, concentration of luminol choose as 2.0×10-7mol mol/L , alkaline concentration of luminol choose as 0.0050 mol /L , concentration of Ag(III) chosen as 2.5×10-5 mol /L and alkaline concentration of Ag(III) was 0.040 mol/L),The detection limit (3σ) was 3.0×10-11 mol L?1, plots of the CL intensity versus concentration of dopamine were linear in the range of 1.0×10-10 ? 4.0×10?8 mol L?1.The regression equation wasI = 22.47 + 1139.42×109 C (r = 0.9996). The RSD (n = 11) was 2.3 % for 1.0×10-8 mol L?1 dopamine.Conclusions: The detection tool that we adopt includes flowing injection technique luminol chemoluminescence system and Ag(III) Complex is more sensitive than the high-efficiency liquid chromatography, fluorometric method, electrochemistry,other flowing injection techniques.The advantages of the detection tool are that simple equipment,cost less,lower detection limit,wider linear range,high speed reaction and reprocessable,so the method provides the technique support for us to study the relationship between catecholamine and trauma and it is more meaningful for us to take measures to prevent diseases about clinical medicine. |
