An Experimental Study On Biochemical Reagent For Battlefield First Aid With Micro Spectrometer

Objective:To meet the first aid needs of battlefield wounded persons (corresponding rate increment of serious injuries like shock, bleeding, infection, multiple organ failure and so on) in the high-technology partial war, and meet the requirements of convenience and automation of first-aid equipments in field war, we plan to develop biochemical detection equipments and matched reagents, which is based on low-voltage-start miniature optic spectrometers and can be used in field war. Detection indexes include hemoglobin, glucose and kalium, sodium, chloridion, carbon dioxide combining power. Combination of these six indexes will form an examining unit for first-aid biochemical functions, and one test gives six results. This research focuses on how to improve the six detection reagents and how to make them meet the requirements of miniature optic spectrometers. And it discusses the feasibility of simultaneous detection, and creates conditions to simplify operation, and cuts report time to fit in field detection, and provides technical support to enhance battlefield first-aid ability.Methods:1. Select a suitable method and reagent for each examining index in the wave length of miniature optic spectrometer (380nm～720nm).2. Make each selected index's liquid detection reagent into dry reagent respectively: select the dry method, and filter the additive categories and concentration for the improvement of reagent appearance and performance.3. Adjust the dried reagent dilution ratio and examining time and adjust and substitute the chromogenic agent, and then improve the element and examining method for the dried reagent.4. Observe the all examining indexes initially and test the performance of dried reagent: make a primary comment on the accuracy, repeatability and stability of the dried reagent. 5. Freeze-dry all examining reagents of the six indexes in self-made detection cup synchronously for the primary application by micro spectrometers, and at the same time evaluate examining reagent performanceperformances, and statistically analyze and deal with the detection results by the software of SPSS 11.0.6. Utilize and evaluate the equipment temperature control system and magnetic mixing system.Results:1. Hemoglobin examining reagent is made up of ferrate potassium cyanide, potassium cyanide, ionic surfactant and so on, and can be freeze-dry directly. Dried reagent has good solvability. When the dilution decreased to 125:1, the dried reagent not only has good solvability, but also gets better linearity range. Coefficient of variation in group of the reagent is 2.22%, and the average interassay variation coefficient is within 3.37%,. The reagent is stable in 3 months.2. Glucose examining reagent is made up of enzyme agent, self-made chromogen TBHBA of 0.5%, and albumin addictive of 10g/L. Increase the dilution multiple of the reagent to 200:1, and cut the reagent examining time to 10min. Coefficient of variation in groupof the reagent accuracy experiment is 2.15%, and the average interassay variation coefficient is within 3.76%. The reagent is stable in 3 months.3. Potassium ion examining reagent is made up of lithium tetraphenylborate proteolytic enzyme, sodium hydroxide, and xylitol addictive of 20g/L. Finished dried reagent has good solubility and reactivity. Coefficient of variation in group of the reagentof the reagent is 2.98%, and the average interassay variation coefficient is within 4.06%. The reagent is stable in 3 months.4. Sodium ion examining reagent is made up ofβ-galactosidase, 0-nitrophenol-galactoside, buffer solution and so on. Finished dried reagent has good solubility. Coefficient of variation in group of the reagentof the reagent is 2.10%, and the average interassay variation coefficientis within 4.34%. The reagent is stable in 3 months.5. Chloride examining reagent is made up of mercury thiocyanate, ferric nitrate, mercury nitrate, and dextran addictive of 40g/L. Adjust the dilution multiple of the reagent to 100:1, and get better examining performanceperformance. Coefficient of variation in group of the reagent of the reagent is 2.10%, and the average interassay variation coefficient is within 3.15%. The reagent is stable in 3 months.6. Examining reagent of carbon dioxide is made up of carbonic anhydrase, indicator, and addictive of polyethylene glycol of 20g/L. The accuracy experiment's bias coefficient the finished dried reagent is 3.27%, and the coefficient of variation in group of the reagent is 3.57%, and average interassay variation coefficient 5.41%. has good solubility and repeatability. The reagent is stable in 3 months.7. According to the freeze-drying method for examining reagents of the six indexes, freeze-dry them in the self-made cup directly, and use uniform diluent, and mix magnetically and heat automatically after loading, and test in designated wave length by the system after incubating, and output the report quickly by computer microprocessing. Examine results show that bias coefficient of each reagent is within 10%, within-batch and inter-batch repeatability (CV%) of each index reagent is less than 5%, except the reagent of carbon dioxide combining power more than 5%. All reagents are stable in 3 months. Relevance between the examining results and dry chemistry analyzer VITRO S-250 is more than 0.97.8. Air incubation system of miniature optic spectrometers can reach 37℃in 3min for reagent incubation, at the same time 92% of mixing system has good mixing status.Conclusions:1. By the improvement of the method and composition of the six first-aid biochemical detection of hexokinase, glucose and kalium, sodium, chloridion, carbon dioxide combining power, we developed the examining dry reagent for miniature optic spectrometers. The reagent is stable within 3 months with accurate and reliable results which can meet basic requirements in clinical examination. The reagent is easy for storage and carrying.2. Performance examining unit formed by the composite examination of multi-project first-aid biochemical indexes, can gain 6 examining results from one examination. It can provide necessary data for the on-site first aid of the wounded, and cut the reporting time which ensures the timely treatment.3. Low-voltage-start miniature optic spectrometers use dry batteries for energy, and can finish automate incubation and mixing without alternating current. The overall equipments have good portability and automation, which can meet the needs of battlefield first aid and on-site quick examination. 4. The development of matched miniature centrifuges can separate examining samples quickly and effectively. The equipments run by dry batteries, which breaks the environmental limitation and realized the purpose of on-site quick examination.In conclusion, the experimental research proves that the biochemical reagent performance for the battlefield first aid is reliable, and the simultaneous examining method is feasible on the basis of miniature optic spectrometers. The overall examining system is suitable for battlefield and can meet the portable and automate needs in battlefield examination, and realizes the purpose to provide quick and reliable information for battlefield first aid.