Study Of Endotoxin Removal In Kidney Dialysis Patient By Amide Membrane Package

Objective: The research gathers the amide membrane package to dialysis patient in vivo, in vitro endotoxin removeing , The counter- seep, the supercarbonate dialysis fluid (the A fluid), 204 dialysis fluids (the B fluid), the pipeline, the albumin, the blood plasma and so on under the different speed of flow, the density, the temperature condition the influence effect which removes to the endotoxin, in order to improve the clinical dialysis process and reduces patient's economical burden.Methods:1, The amide membrane package of preparation: Prepares Nylon-NH-CHOH-(CH₂)₃- CHOH-NH-(CH₂)₆-NH-CH₂-CHOH-CH₂-Histidin (histidine) ,Causes agarose to hand over the association and to activate, the connection gap arm(hexanediamine), causes product AH-SepharoseCL-4B. Again causes the genin solid truncation and the medium regeneration.2, Disinfection method:(1)Glass appliance disinfection: 300℃under disinfects 40 minutes in the silicon-controlled rectifier electric stove, after natural cooling, the same day to use.(2)Pipeline and membrane separator disinfection: The young rubber tube uses boils 15 minutes disinfection methods.3, Standard endotoxin solution configuration: Matches the different density the endotoxin working standard with the endotoxin inspection water .4, Adsorption method:(1)Static adsorption method : takes the dry weight 15mg membrane, sets the endotoxin inspection water used to 2ml, the interrupted vibration, lays aside for 2 hours, attracts the serum, the determination endotoxin density, through adsorption around density computation removeing rate.(2) Dynamic adsorption law: Gathers the amide membrane 30 to load in the membrane separator, with wriggles pumps to contains the endotoxin solution to flow by four kind of speeds of flow afterthe membrane separator, a collection membrane package of around sample, fixed time, decides under the warm condition, the determination endotoxin content, determined the membrane package removes the situation to the endotoxin.5, The membrane package to the dialysis fluid in endotoxin removeing: (1) A, B fluid ion content determination: Determines in the solution with the calcium purplish red element indicator CaCl₂; Determines in the solution with the Erichrome black T indicator MgCl₂6H₂O; Determines in the solution with the tetrabromophenol sulfonphthalein indicator KCl; Instructs in the fluid determination solution with the silver acetate NaCl; Determines in the solution with the phenolphthalein indicator the sodium acetate; Uses in the optical rotation law determination solution the glucose content. (2) membrane package to the dialysis fluid in the fixed ion, the pH condition eliminates the endotoxin: With reagent law examination membrane package to dialysis fluid in different speed of flow (5, 10, 15, 20ml/min) under condition to endotoxin removeing rate and various ions change situation.6, Pipeline endotoxin determination method: According to the national standard, soaks raises the medium, the bacterium endotoxin inspection water used, pipeline and the vessel class appliance soaks the appliance cavity with the bacterium endotoxin inspection water used, does not surpass for the trying storage for 2 hours.7, The method of removing the endotoxin with the membrane package to dialogue protein in: The person with the albumin take 10 time of dilutions, wraps in the speed of flow after the membrane as 5ml/min, under 25℃conditions has filtered, the examination albumin dams the current the situation, and around the endotoxin removes the situation.8,The method of removing the endotoxin with the membrane package to the blood plasma in: Takes the patient blood plasma, by 10 time of dilutions, contrasts the membrane package of around endotoxin content under the different speed of flow, computation receiving rate and the blood ion change, after and examination membrane albumin content.9,Endotoxin quantitative determination (1) endotoxin content standard curve preparation: Carries on the determination density using the BET-16 bacterium endotoxin cryoscope is 7.0, 1.75, 0.875, 0.438, 0.219, 0.109, 0.055, 0.014EU/ml endotoxin standard series watery solution. The instrument may act according to agglutinates the time logarithm (lgT) (lgC) to formulate the standard curve to the bacterium endotoxin density logarithm. (2) reagent sensitivity reexamines the experiment: Before eachbatch of reagents primary use, must carry on to its indication sensitivity reexamines. Qualified may supply the bacterium endotoxin examination.Results:1. When the speed of flow respectively is 5ml/min, 10ml/min, 15ml/min, 20ml/min, the endotoxin elimination rate of counter-seep solution (including endotoxin 4.000 EU/ml) respectively is 82.3%,80.6%,79.8%,79.2%. When speed of flow is 5ml/min elimination rate is highest, along with the speed of flow increase, the endotoxin elimination rate has the drop tendency. When the endotoxin density is 4.000 EU/ml blood plasmas under speed of flow 5ml/min, 10ml/min, 15ml/min, 20ml/min condition, the endotoxin elimination rate respectively is 87.6%,86.1%,83.4%,79.3%。When 5ml/min removes the effect to be best, along with the speed of flow increase, the endotoxin elimination rate has the drop tendency.2. Under the condition of temperature respectively is 15℃, 25℃, 37℃, the endotoxin elimination rate respectively is 88.8%,89.5%,89.9%,Obviously the influence of temperature to the endotoxin elimination is not obvious , along with the temperature ascension, the elimination rate has the rise slightly.3. When the speed of flow is 5ml/min, the temperature for 25℃To does not contain in the albumin (HAS) water-soluble fluid, the endotoxin elimination rate were 89.6% for static adsorption and 98.4% for dynamic adorption. Joins in the HAS water-soluble fluid the endotoxin elimination rate were 81.2% for static adsorption and 86.6% for dynamic adorption. Obviously the albumin has affected in the water-soluble fluid the endotoxin elimination. Protein returns-ratios were 85.8% for static method and 93.8% for dynamic adsorption, The adsorbability of the amide to HAS is very small .4. Pipeline which the patient hemodialysis liquid flow passes through, keeps warm for 1 hour according to the national standards under condition of 37℃. collection filtrate, the endotoxin value is: The vein manages 5.36EU; The artery manages 3.855EU. Conforms to the national standards.Conclusion: The amide membrane package, take the histidine as the genin, synthesized take agarose as the matrix membrane medium, and has the certain length the gap arm. This medium may effectively remove the endotoxin of liquid and so on in water, dialysis fluid, blood plasma and albumin, also may the higher retention each kind of matter effective component.