疫苗中蛋白质检测方案(定氮仪)

APPLICATION NOTEF&F-D-002-2017/A1 N/PROTEIN DETERMINATION IN VACCINEDUMAS COMBUSTION METHOD N/Protein Determination inVaccines according to the Dumascombustion method Reference： VELP Internal Procedure Tested with VELP Scientifica NDA 701 Dumas Nitrogen Analyzer (Code F30800070) Introduction A vaccine contains a part of a germ (bacteria or virus) that is called an antigen. The antigen has already been killed ordisabled before it's used to make the vaccine， so it can't make you sick. Antigens are substances， often a protein， thatstimulate the body to produce an immune response to protect itself against attacks from future actual disease exposure.In addition， vaccines contain other ingredients that make them safer and more effective， including preservatives， adjuvants，additives and residuals of the vaccine production process. Vaccines have reduced and， in some cases， eliminated many diseases that killed or severely disabled people just a fewgenerations ago. Vaccines， like any medication， are continually monitored for safety. Protein Determination in two different vaccine solutions The Dumas method starts with a combustion furnace (CF) to burn thesample， obtaining elemental compounds. Water is removed by a first physical trap (WT1-DriStep TM)， placed after thecombustion， and a second chemical one (WT2). Between the two， theelemental substances passed through a reduction furnace (RF). The auto-regenerative CO2absorbers (CO2) let pass only the elementalnitrogen that is detected by the LoGas TM innovative Thermal ConductivityDetector (TCD) with no requirement for a reference gas. The NDA 701 is controlled via PC through the intuitive DUMASoftTM. NDA 701 Preliminary Operations (daily) Follow the operating manual to start the NDA 701 and check that the following parameters are set： Temperature Combustion reactor (Code A00000158)： 1030°C Temperature Reduction reactor (Code A00000226)：650℃ Flow rate MFC1 He： 190 ml/min Flow rate MFC2 He： 220 ml/min Condition the system by testing 2 EDTA standard (Code A00000149) and 3 to 5 empty tin foils (Code A00000153) asCheck up. Verify the calibration curve with one or more tests as Standard by testing the same standard used for thecurve creation. Sample Preparation Two liquid vaccine samples have been analyzed： Dipthteria and Tetanus. Samples have been kept in vials at refrigeratedtemperature until the test. In order to homogenize the sample， shake the vial through VELP Vortex TX 4 (Code F202A0270). Put into the tin foil 50-70 mg of Super-Absorbent Powder (CodeA00000317). Pour into the tin foil around 200 pl(200-220 mg) of vaccine solution using a micropipette. Carefully close the tin foil， obtaining a capsule. Load the capsule into the autosampler. Analysis Procedure Fill the following fields in the database： Sample name， Weight， Method， Sample type， Calibration numberCreate a new customizable method with the following parameters： Protein factor： 6.25 O2 flow rate： 300 ml/min O2 factor： 0.7 ml/mg Press to start the analysis. Analysis time： from 3 minutes for one run. Typical Results on Vaccine samples Results have been obtained with the following calibration curve： in a range of 0 - 1.7 mg N with 5 measurements of Riceflour standard (N%=1.38) (Code A00000235). The data obtained are included in the tolerance admitted by the Rice flourcertificate. The table below shows the nitrogen/protein results， obtained by the DumasoftTM. Sample Sample quantity (mg) Nitrogen % Protein % Diphteria 220.90 0.100 0.628 217.00 0.099 0.619 216.90 0.091 0.570 219.00 0.100 0.624 215.70 0.098 0.612 Average±SD% 0.098±0.004 0.611±0.023 Tetanus 199.50 0.127 0.791 201.80 0.131 0.816 206.80 0.131 0.821 204.30 0.130 0.811 210.90 0.130 0.814 Average ± SD% 0.130±0.002 0.811±0.012 Protein Factor： 6.25 Conclusion Nowadays the attention on safety of vaccines has increased due to their potential adverse reactions after injection.Nitrogen/protein analysis according to the Dumas principle with the combustion apparatus is a credible alternative to theacid digestion method according to Kjeldahl. NDA 701 ensure reliable results in a easy and fast way. Despite low sample weights， excellent reproducibility are possibleand the data confirm the complete combustion of the sample with no memory effect observed. With high productivity and non-stop performances， NDA 701 Dumas combustion apparatus is ideal for high throughput，being fully automated and requiring just 3-4 minutes per analysis. Copyright C VELP Scientifica. All rights reserved.No part of this publication may be reproduced or transmitted in any form or for any purpose without the express permission of VELP.VELP Scientifica， Italy-Tel： + Fax： +ww.velp.com 利用VELP NDA 701杜马斯定氮技术分析疫苗产品中的蛋白质: 含量疫苗包含称为抗原的一部分细菌（细菌或病毒）。 该抗原在制作疫苗之前已经被杀死或禁用，因此不会使您生病。 抗原是一种物质，通常是一种蛋白质，可以刺激人体产生免疫反应，从而保护自身免受未来实际疾病的侵袭。 此外，疫苗还包含使其更安全有效的其他成分，包括防腐剂，佐剂，添加剂和疫苗生产过程中的残留物。疫苗已经减少，并且在某些情况下，消除了仅几代人以前就导致许多人死亡或重度残疾的疾病。像任何药物一样，疫苗也要不断进行安全性质控。