CN1173161C - A micro-sampling device for an on-line detection system of a biological reaction process - Google Patents
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Abstract
一种生物反应过程在线检测系统的取样稀释装置,具体涉及微量取样装置和级联稀释装置。取样稀释采用连续流动、间歇测量的方式。微量取样装置主要在取样探头末端放置一陶瓷膜过滤件,反应液在罐中膜件中原位过滤,错流渗透,渗透液由膜外导流夹套螺纹导流引出,实现从罐外对罐内自动微量取样。级联稀释装置由带溢流腔的垂直三级级联稀释板为主所组成,流体流动依赖流体自身重力,靠有机材料的微全流路结构,使样品和稀释液的流动保持稳定。本装置样品取用量很少,不会影响生物反应过程的优化控制,稀释比容易控制,操作方便、灵活,制造简单,价格低廉。主要构件均可制作为标准件,易于清洗和更换。样品代表性好,适用于生物反应过程在线检测和微全分析(μ-TAS)的应用。
The invention relates to a sampling dilution device of an online detection system of a biological reaction process, in particular to a micro sampling device and a cascade dilution device. Sampling dilution adopts the method of continuous flow and intermittent measurement. The micro-sampling device mainly places a ceramic membrane filter at the end of the sampling probe. The reaction liquid is filtered in situ in the membrane in the tank, cross-flow infiltrates, and the permeate is led out by the diversion jacket thread outside the membrane, so as to realize the transfer from the outside of the tank to the tank. Internal automatic micro sampling. The cascade dilution device is mainly composed of vertical three-stage cascade dilution plates with overflow chambers. The fluid flow depends on the gravity of the fluid itself, and the micro-full flow path structure of organic materials keeps the flow of samples and diluents stable. The device takes very little sample, does not affect the optimal control of the biological reaction process, is easy to control the dilution ratio, is convenient and flexible to operate, simple to manufacture, and low in price. The main components can be made as standard parts, which are easy to clean and replace. The sample is well represented and is suitable for the application of on-line detection of biological reaction process and micro-total analysis (μ-TAS).
Description
技术领域technical field
本发明涉及一种生物反应过程在线检测系统的取样稀释装置,与生物传感器和微机相配套以实现生物反应过程中物质浓度的在线检测,具体涉及包括微量取样装置和级联稀释装置。The invention relates to a sampling and dilution device for an online detection system of a biological reaction process, which is matched with a biosensor and a microcomputer to realize the on-line detection of substance concentration in the biological reaction process, and specifically relates to a micro sampling device and a cascade dilution device.
现有技术current technology
当今的发酵工业除了传统酿造之外,已向抗生素、氨基酸、核酸等的发酵生产方向发展。近年来,正在研究利用微生物重组和细胞培养等技术生产生物活性物质。伴随着上述的发展过程,发酵过程的控制和最优化也已从靠经验管理向用微机自动化管理的方向发展。In addition to traditional brewing, today's fermentation industry has developed towards the fermentation production of antibiotics, amino acids, nucleic acids, etc. In recent years, the use of technologies such as microbial recombination and cell culture is being studied to produce biologically active substances. Along with the above-mentioned development process, the control and optimization of the fermentation process has also developed from experience management to computer automation management.
各种各样的物理和化学传感器广泛地用于过程的控制中。检测温度、pH、溶解氧、一氧化碳、压力、流量、搅拌功能等参数的传感器和微机连接,构成了用于生化和发酵过程中的控制系统。但是它只能间接地对生化反应进行控制,并不能直接了解复杂的生化过程中物质的变化情况。而生物传感器解决了许多过去难以进行的物质分析,可以测定直接代表发酵过程进行状态的原料和发酵产物的浓度。经过20多年的研究开发,生物传感器已形成一个独特的门类,与物理传感器和化学传感器并列起来,看作是传感器的第三个类别。Various physical and chemical sensors are widely used in the control of processes. Sensors for detecting parameters such as temperature, pH, dissolved oxygen, carbon monoxide, pressure, flow, and stirring functions are connected with a microcomputer to form a control system for biochemical and fermentation processes. But it can only indirectly control biochemical reactions, and cannot directly understand the changes of substances in complex biochemical processes. The biosensor solves many substances that were difficult to analyze in the past, and can measure the concentration of raw materials and fermentation products that directly represent the state of the fermentation process. After more than 20 years of research and development, biosensors have formed a unique category, juxtaposed with physical sensors and chemical sensors, and are regarded as the third category of sensors.
生物传感器具有专一性好、酶可以多次重复使用、分析速度快、准确度高、操作系统比较简单等特点,为微机直接控制生物反应过程准备了可行的传感器条件。但是,至今还未见报道能在一般生物反应过程中实现在线检测生物、化学量的分析系统。生物、化学量无法实现在线检测,究其根源是生物传感器与复杂的生物反应过程的检测接口技术无法过关,这已成为生物传感器实现在线检测的“瓶颈”技术。Biosensors have the characteristics of good specificity, enzymes can be reused many times, fast analysis speed, high accuracy, and relatively simple operating system, which prepares feasible sensor conditions for direct control of biological reaction processes by microcomputers. However, no analysis system has been reported that can realize on-line detection of biological and chemical quantities in the general biological reaction process. Biological and chemical quantities cannot be detected online. The root cause is that the detection interface technology between biosensors and complex biological reaction processes cannot pass the test. This has become the "bottleneck" technology for biosensors to achieve online detection.
生物反应过程在线检测系统示意图如图1所示,包括微量取样器、稀释器、微型生物传感器组、生物工程数据处理分析软件和微机。而微量取样器和稀释器正是本发明要解决的技术问题。The schematic diagram of the online detection system for the biological reaction process is shown in Figure 1, including a micro sampler, a diluter, a micro biosensor group, bioengineering data processing and analysis software and a microcomputer. And the micro sampler and the diluter are the technical problems to be solved in the present invention.
在科研工作中,实验人员采用全自动小型发酵罐进行发酵工艺条件的优化,普遍使用2-30L体积的小罐,而取样体积高达15ml,多次取样造成发酵液体积减少,对发酵过程带来较大的影响,会导致过程优化的误差增大。所以迫切需要设计一种微量取样装置,减少取样体积,尽量降低取样对检测的干扰。In scientific research work, experimenters use fully automatic small fermenters to optimize the fermentation process conditions. Small tanks with a volume of 2-30L are generally used, and the sampling volume is as high as 15ml. Multiple samplings cause the volume of the fermentation broth to decrease, which brings great harm to the fermentation process. A larger impact will lead to an increase in the error of process optimization. Therefore, it is urgent to design a micro-sampling device to reduce the sampling volume and minimize the interference of sampling to detection.
目前已有的稀释装置,如机械臂,流动注射分析在线稀释技术,为高浓度样品的在线测定提供了有效的方法之一;然而需要时间控制器、计算机控制阀和精度较高的蠕动泵,因而机械加工要求高,价格昂贵,稀释比的控制不灵活,严重影响了生物传感器在生物反应过程的在线检测应用。Currently existing dilution devices, such as robotic arms and flow injection analysis online dilution technology, provide one of the effective methods for online determination of high-concentration samples; however, time controllers, computer-controlled valves and high-precision peristaltic pumps are required. Therefore, the requirements for mechanical processing are high, the price is expensive, and the control of the dilution ratio is inflexible, which seriously affects the online detection application of biosensors in the biological reaction process.
发明内容Contents of the invention
(1)要解决的技术问题(1) Technical problems to be solved
提供一种生物反应过程在线检测系统的微量取样装置,要求解决从罐外对发酵罐内进行微量取样的技术问题,既克服了生物传感器不耐热的特点,以及克服氧浓度和温度对生物电极的影响,又减少了取样体积,尽可能降低取样对检测的干扰,保持在线连续取样、间歇测量的需要。Provide a micro-sampling device for the on-line detection system of the biological reaction process, which requires solving the technical problem of micro-sampling the fermentation tank from the outside of the tank, which not only overcomes the heat-resistant characteristics of the biosensor, but also overcomes the impact of oxygen concentration and temperature on the bioelectrode. The impact of the sample volume is reduced, the interference of sampling to the detection is reduced as much as possible, and the need for continuous online sampling and intermittent measurement is maintained.
还提供一种生物反应过程在线检测系统的级联稀释装置,要求解决可以进行在线连续稀释的技术问题。采用缓冲液对样品进行几十到几百倍的稀释,灵活控制稀释率,以达到生物传感器检测的线性范围。稀释的另一作用是调节样品pH值,使之符合传感器工作pH值,减少干扰。Also provided is a cascade dilution device of an online detection system for a biological reaction process, which requires solving the technical problem that online serial dilution can be performed. The buffer solution is used to dilute the sample tens to hundreds of times, and the dilution rate is flexibly controlled to achieve the linear range of biosensor detection. Another function of dilution is to adjust the pH value of the sample to match the working pH value of the sensor and reduce interference.
(2)技术方案(2) Technical solution
微量取样装置micro sampling device
图2为微量取样装置结构示意图,它由一个取样探头和样品流量控制部分组成。探头末端为膜过滤部分,另一端为渗透液引出管和与罐连接密封部分。取样探头由探头圆柱体、膜过滤件、膜外导流夹套、导流接插件、引出管、套管组成。流量控制部分由针状阀、计量阀共同进行流量控制。取样探头、针状阀、计量阀之间由引出管连接。Fig. 2 is a schematic structural diagram of a micro-sampling device, which consists of a sampling probe and a sample flow control part. The end of the probe is the membrane filter part, and the other end is the permeate outlet pipe and the sealing part connected with the tank. The sampling probe is composed of a probe cylinder, a membrane filter, a diversion jacket outside the membrane, a diversion connector, a lead-out tube, and a casing. The flow control part is jointly controlled by the needle valve and the metering valve. The sampling probe, the needle valve and the metering valve are connected by an outlet pipe.
膜过滤件采用陶瓷膜件,膜过滤件为管状,膜层在内壁,支撑层在外,内径为6-10mm,外径为10-14mm。膜过滤件长8-12mm,膜孔径0.1-0.2μm。The membrane filter adopts a ceramic membrane, and the membrane filter is tubular, with a membrane layer on the inner wall and a support layer on the outside, with an inner diameter of 6-10mm and an outer diameter of 10-14mm. The length of the membrane filter is 8-12mm, and the membrane pore diameter is 0.1-0.2μm.
膜外导流夹套,在垂直放置的探头圆柱体的末端上水平钻孔,即为膜外导流夹套。在膜外导流夹套内壁做成螺纹状,用于导流,螺纹大径和小径之差小于2mm。膜过滤件置于探头圆柱体的膜外导流夹套钻孔中,膜外导流夹套内螺纹和膜过滤件外壁接触,膜外导流夹套和膜过滤件采用过盈配合,配合处两端用硅胶密封。探头圆柱体选用聚偏二氟乙烯或聚四氟乙烯材质。The outer membrane diversion jacket is horizontally drilled on the end of the vertically placed probe cylinder, which is the outer membrane diversion jacket. The inner wall of the diversion jacket outside the membrane is threaded for diversion, and the difference between the major diameter and the minor diameter of the thread is less than 2mm. The membrane filter is placed in the drill hole of the outer membrane diversion jacket of the probe cylinder. The inner thread of the outer membrane diversion jacket is in contact with the outer wall of the membrane filter. The outer membrane diversion jacket and the membrane filter adopt an interference fit. Both ends are sealed with silicone. The probe cylinder is made of polyvinylidene fluoride or polytetrafluoroethylene.
在探头圆柱体与套管相接的一端圆面中间开一锥形头圆柱形小孔,小孔圆柱部分有内螺纹,小孔与导流夹套中部相通,在小孔内置导流接插件。导流接插件为一带锥形头的小圆柱,小圆柱的圆柱部分设外螺纹,导流接插件和探头圆柱体以螺纹配合,导流接插件的锥形头紧靠膜外导流夹套以减少死体积。在导流接插件柱锥体中间开孔,插入引出管。导流接插件选用聚四氟乙烯材质。A small cylindrical hole with a conical head is opened in the middle of the circular surface at the end where the probe cylinder and the casing are connected. The cylindrical part of the small hole has internal threads. The small hole communicates with the middle part of the diversion jacket, and a diversion connector is built in the small hole. . The diversion connector is a small cylinder with a tapered head, the cylindrical part of the small cylinder is provided with external threads, the diversion connector and the probe cylinder are threaded, and the conical head of the diversion connector is close to the diversion jacket outside the membrane to reduce dead volume. Open a hole in the middle of the cone of the diversion connector, and insert the outlet tube. The diversion connector is made of polytetrafluoroethylene.
引出管为塑料弹性管,选用聚硅酮弹性管或聚偏二氟乙烯弹性管。The lead-out tube is a plastic elastic tube, and a polysilicone elastic tube or a polyvinylidene fluoride elastic tube is selected.
套管一头设有内螺纹和探头圆柱体一端的外螺纹以螺纹配合。套管选用不锈钢材质,以保护引出管。One end of the bushing is provided with an internal thread and the external thread at one end of the probe cylinder is threadedly matched. The casing is made of stainless steel to protect the outlet pipe.
取样采用错流过滤渗透技术,用在原位的过滤器让反应液流过膜件,直接在罐中过滤,利用膜件实现发酵液与固体的分离,膜件外采用螺纹导流,引出滤液后直接处理,连续过滤。用阀控制渗透液流量,稳定状态时膜的扩散量与渗透液流量相等,使渗透液连续引出。每隔一定时间取渗透液作为测量样品,即间歇测量,不作为测量样品的渗透液直接排掉。Sampling adopts cross-flow filtration and osmosis technology. The in-situ filter is used to allow the reaction liquid to flow through the membrane and filter directly in the tank. The membrane is used to separate the fermentation liquid from the solid. The outside of the membrane is guided by threads to lead out the filtrate. After processing directly, continuous filtration. The permeate flow is controlled by a valve, and the diffusion amount of the membrane is equal to the permeate flow in a steady state, so that the permeate is continuously drawn out. The permeate is taken as a measurement sample at regular intervals, that is, intermittent measurement, and the permeate that is not used as a measurement sample is directly discharged.
设计参数为:渗透液流量0.4ml/min,滞后时间小于1min,取样器管路体积≤0.7ml,取样量10-1000μl/min。The design parameters are: permeate flow rate 0.4ml/min, lag time less than 1min, sampler pipeline volume ≤0.7ml, sampling volume 10-1000μl/min.
级联稀释装置cascade dilution device
图3在线检测系统中三级级联装置的接口流程示意图。级联稀释装置由缓冲液溢流部分和带溢流腔的垂直三级级联稀释板组成。缓冲液溢流部分包括缓冲液储罐、恒流泵、三个缓冲液溢流槽,缓冲液流量由缓冲液流路上的缓冲液流量调节阀控制。图4为带溢流腔的垂直三级级联稀释板的结构示意图。结构为:由带溢流腔的样品流路和缓冲液流路汇合成稀释混合流路,作为一级稀释器,将三个一级稀释器垂直串联组成三级级联稀释板,将第一级稀释混合流路和第二级溢流腔接通,将第二级稀释混合流路和第三级溢流腔接通。将第一级稀释混合液作为第二级的样品继续稀释,将第二级的稀释混合液作为第三级的样品再稀释,将第三级的稀释混合液作为检测样品进行检测。溢流腔底部设置有排液阀,溢流腔顶部设置有通气孔。样品流量由样品流路上的样品流量调节阀控制,级联稀释板选用有机材料,如聚偏二氟乙烯或聚四氟乙烯。流体流路采用与微全分析系统(μ-TAS)相适应的微全流结构,充分缩短样品的停留时间,但又不致形成液滴,流体流动依赖流体自身的重力,既充分保持流体的静压,尽可能少产生动压,所有操作必须是连续稳定流动、连续取样、连续稀释、采用间歇测量。Figure 3 is a schematic diagram of the interface flow of the three-stage cascade device in the online detection system. The cascade dilution device consists of a buffer overflow section and a vertical three-stage cascade dilution plate with an overflow chamber. The buffer overflow part includes a buffer storage tank, a constant flow pump, and three buffer overflow tanks, and the buffer flow is controlled by a buffer flow regulating valve on the buffer flow path. Fig. 4 is a schematic structural diagram of a vertical three-stage cascaded dilution plate with an overflow chamber. The structure is: the sample flow path with an overflow chamber and the buffer flow path are merged into a dilution mixing flow path, as a primary diluter, three primary diluters are vertically connected in series to form a three-stage cascade dilution plate, and the first The first-stage dilution mixing flow path is connected to the second-stage overflow chamber, and the second-stage dilution mixing flow path is connected to the third-stage overflow chamber. The first-level diluted mixed solution is used as the second-level sample to continue diluting, the second-level diluted mixed solution is used as the third-level sample for further dilution, and the third-level diluted mixed solution is used as the test sample for detection. A drain valve is arranged at the bottom of the overflow chamber, and a vent hole is arranged at the top of the overflow chamber. The sample flow is controlled by the sample flow regulating valve on the sample flow path, and the cascade dilution plate is made of organic materials, such as polyvinylidene fluoride or polytetrafluoroethylene. The fluid flow path adopts a micro total flow structure compatible with the micro total analysis system (μ-TAS), which fully shortens the residence time of the sample, but does not form droplets. The fluid flow depends on the gravity of the fluid itself, which not only fully maintains the static state of the fluid Pressure, generate as little dynamic pressure as possible, all operations must be continuous and stable flow, continuous sampling, continuous dilution, and intermittent measurement.
流体流路管径为0.1-1mm,稀释混合流路长度为5-10cm,溢流腔腔深为3-6mm。单级稀释比控制在4-10,稀释误差不大于1%,三级稀释比最高为1000,稀释误差不大于2.5%。The diameter of the fluid flow path is 0.1-1mm, the length of the dilution and mixing flow path is 5-10cm, and the depth of the overflow cavity is 3-6mm. The single-stage dilution ratio is controlled at 4-10, and the dilution error is not greater than 1%, and the third-stage dilution ratio is up to 1000, and the dilution error is not greater than 2.5%.
(3)有益效果(3) Beneficial effect
本发明设计的微量取样装置、三级级联稀释装置最大的特点是微量取样、微量稀释,以供微型生物传感器的微量检出,其样品用量为10-100μL/min,发酵100小时,连续取样的样品损失量也仅为60-600ml,能满足小型发酵罐取样检测的要求。The biggest feature of the micro-sampling device and three-stage cascade dilution device designed in the present invention is micro-sampling and micro-dilution for micro-detection of micro-biosensors. The sample consumption is 10-100 μL/min, fermented for 100 hours, and continuously sampled The sample loss is only 60-600ml, which can meet the requirements of sampling and testing of small fermenters.
微量取样装置以自动取样方式取代手动取样方式,采用错流过滤技术,利用膜件实现发酵液与固体的分离,膜件外侧采用螺纹导流,只要反应液在膜件管内流过,就会由滤膜层渗透流入夹套螺纹内,形成错流渗透,实现液体的先进先出,即有反应液流进膜件管内,就会见到渗透液引出。The micro-sampling device replaces the manual sampling method with an automatic sampling method, adopts cross-flow filtration technology, and uses a membrane to separate the fermentation liquid from the solid. The filter membrane layer permeates into the jacket thread, forming a cross-flow permeation, and realizes the first-in-first-out liquid, that is, the reaction liquid flows into the membrane tube, and the permeate is drawn out.
采用在原位的过滤器,直接在罐中过滤,取出滤液作为样品,其优点是:不增加罐的复杂性,在罐安装探头处装过滤膜件,不会增加染菌机会,膜件和管路易于清洗,且均为标准件,可以及时更换。The in-situ filter is used to filter directly in the tank, and the filtrate is taken out as a sample. The advantages are: the complexity of the tank is not increased, and the filter membrane is installed at the probe of the tank, which will not increase the chance of bacterial contamination. The membrane and The pipelines are easy to clean, and all are standard parts, which can be replaced in time.
级联稀释装置设计了依赖流体自身重力的运行,即溢流来控制稀释样品和缓冲液流量的稳定性,无需时间控制器、计算机控制阀和高精度的蠕动泵,操作方便、灵活,适合生物反应过程在线检测的样品稀释要求。其三级稀释和流体流路的微全流结构,使样品形成稳定的连续流动,稀释率不随样品或缓冲液流量的改变而改变或变化很小。The cascade dilution device is designed to operate relying on the gravity of the fluid itself, that is, overflow to control the stability of the diluted sample and buffer flow, without the need for a time controller, computer-controlled valve and high-precision peristaltic pump. It is easy to operate and flexible, and is suitable for biological Sample dilution requirements for online detection during the reaction process. Its three-stage dilution and the micro-full-flow structure of the fluid flow path make the sample form a stable continuous flow, and the dilution rate does not change or changes little with the change of the sample or buffer flow.
在线检测系统采用连续自动取样、稀释、间歇测量的方法。连续取样稀释的流程是让样品依此流径取样管路和稀释管路,样品处于连续流动,其流量容易控制,不需要排空死样,且记忆效应小,对样品代表性影响小。连续取样、稀释系统制造简单、廉价、样品代表性好,非常适合于在线分析检测用。可进一步降低生物反应控制成本,提高生物传感器检测的精度。The online detection system adopts the methods of continuous automatic sampling, dilution and intermittent measurement. The process of continuous sampling and dilution is to let the sample flow through the sampling pipeline and the dilution pipeline in this way. The sample is in a continuous flow, and its flow rate is easy to control. There is no need to empty the dead sample, and the memory effect is small, which has little influence on the representativeness of the sample. The continuous sampling and dilution system is simple to manufacture, cheap, and has good sample representation, and is very suitable for on-line analysis and detection. The cost of biological reaction control can be further reduced, and the detection accuracy of the biosensor can be improved.
附图说明Description of drawings
图1生物反应过程在线检测系统示意图。Fig. 1 Schematic diagram of online detection system for biological reaction process.
图2微量取样装置结构示意图。(a)微样取样装置正视示意图:(b)取样探头部分侧视示意图。Figure 2 Schematic diagram of the structure of the micro-sampling device. (a) Schematic diagram of the front view of the micro-sampling device; (b) Schematic diagram of the side view of the sampling probe.
图3在线检测系统中三级级联装置的接口流程示意图。Figure 3 is a schematic diagram of the interface flow of the three-stage cascade device in the online detection system.
图4带溢流腔的垂直三级级联稀释板的结构示意图。Fig. 4 Schematic diagram of the structure of a vertical three-stage cascaded dilution plate with an overflow chamber.
附图中1探头圆柱体,2膜过滤件,3膜外导流夹套,4导流接插件,5引出管,6套关,7针状阀,8计量阀,9溢流腔,10样品流路,11样品流量调节阀,12排液阀,13通气孔,14缓冲液流量调节阀,15缓冲液流路,16稀释混合流路。In the accompanying drawings, 1 probe cylinder, 2 membrane filter, 3 membrane outer diversion jacket, 4 diversion connector, 5 outlet pipe, 6 set of valves, 7 needle valve, 8 metering valve, 9 overflow chamber, 10 Sample flow path, 11 sample flow regulating valve, 12 drain valve, 13 air vent, 14 buffer flow regulating valve, 15 buffer flow path, 16 dilution mixing flow path.
具体实施方式Detailed ways
实施例1Example 1
微量取样装置:膜过滤件采用某公司的陶瓷膜件。膜件为管状,膜层在内壁,支撑层在外,内径d1=8mm,外径d2=12mm。过滤时压强降约为10KPa,膜孔径为0.2μm,要求滤液流量为0.4ml/min,取膜件长为10mm,根据相关公式计算,能满足渗透液流量要求。Micro-sampling device: The membrane filter adopts the ceramic membrane of a certain company. The membrane is tubular, the membrane layer is on the inner wall, the support layer is outside, the inner diameter d1=8mm, and the outer diameter d2=12mm. The pressure drop during filtration is about 10KPa, the membrane pore size is 0.2μm, the filtrate flow rate is required to be 0.4ml/min, and the length of the membrane piece is 10mm. According to the calculation according to the relevant formula, it can meet the permeate flow requirement.
实施例2Example 2
膜过滤件的膜孔径为0.1μm,其余条件同实施例1,也能满足渗透液流量要求。The membrane pore diameter of the membrane filter element is 0.1 μm, and other conditions are the same as those in Example 1, which can also meet the requirement of the flow rate of the permeate.
实施例3Example 3
三级级联稀释板流体流路总长为90mm,流路管径为1mm,则总体积为0.09ml,以0.1ml/min流量考虑其停留时间仅为0.9min,取样10-1000L/min,单级稀释比控制在4-10,单级稀释误差不大于1%,三级稀释比最高为1000,稀释误差不大于2.5%。样品用葡萄糖,其原始浓度为278.54g/l,进行三级稀释试验,其稀释精度测试结果为:The total length of the three-stage cascaded dilution plate fluid flow path is 90mm, and the flow path diameter is 1mm, so the total volume is 0.09ml. Considering the flow rate of 0.1ml/min, the residence time is only 0.9min, and the sampling rate is 10-1000L/min. The level dilution ratio is controlled at 4-10, the single-level dilution error is not greater than 1%, the third-level dilution ratio is up to 1000, and the dilution error is not greater than 2.5%. Glucose was used for the sample, and its original concentration was 278.54g/l, and a three-stage dilution test was carried out, and the results of the dilution precision test were:
表1 三级级联稀释精度的测试结果 Table 1 Test results of three-stage cascaded dilution accuracy
一级稀释 二级稀释 三级稀释 总稀释比 误差率% Primary Dilution Secondary Dilution Tertiary Dilution Total Dilution Ratio Error Rate %
平均浓度 52.23 10.89 2.45Average concentration 52.23 10.89 2.45
稀释倍数 5.33 4.80 4.44 113.69 0.6Dilution factor 5.33 4.80 4.44 113.69 0.6
平均浓度 47.42 6.65 0.94Average concentration 47.42 6.65 0.94
稀释倍数 5.87 7.13 7.07 296.32 1.2Dilution factor 5.87 7.13 7.07 296.32 1.2
平均浓度 28.83 3.30 0.45Average concentration 28.83 3.30 0.45
稀释倍数 9.66 8.74 7.33 618.98 1.8Dilution factor 9.66 8.74 7.33 618.98 1.8
平均浓度 28.83 2.88 0.28Average concentration 28.83 2.88 0.28
稀释倍数 9.66 10.00 10.35 1000 2.5Dilution factor 9.66 10.00 10.35 1000 2.5
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