WO2009135334A1

WO2009135334A1 - Process and apparatus for measuring the concentration of composition during fermentation process

Info

Publication number: WO2009135334A1
Application number: PCT/CN2008/000900
Authority: WO
Inventors: 蔡浩原; 杨宏伟; 郭旻; 贺伯特·格里布; 卓越; 库特·贝腾豪森
Original assignee: 西门子公司
Priority date: 2008-05-05
Filing date: 2008-05-05
Publication date: 2009-11-12

Abstract

An apparatus (100) for measuring the concentration of composition during fermentation process comprises a sample line (104), a reagent line (105), the configuration of which is so designed that the both lines at least partly overlap each other to form a chamber, a dialysis membrane (103) mounted between the sample line (104) and the reagent line (105), allowing the sample to be analysed in the fermentation liquor sample to diffuse from said sample line (104) to said reagent line (105), to make said sample to be analysed and said reagent reaction, and an operating device (114) mounted in said reagent line to measuring optical absorption degree of reagent after response. A process for measuring the concentration of composition during fermentation process can be finished only by a small quantity of fermentation liquor sample and reagent.

Description

Apparatus and method for measuring component concentration in a fermentation process

The present invention relates to the field of monitoring of fermentation processes, and more particularly to a device for measuring the concentration of components in a fermentation process and a method for measuring the concentration of components in a fermentation process. Background technique

Fermentation is a very important process in the field of biochemistry, especially in the pharmaceutical analysis, food, beverage, energy and environmental industries. In order to optimize the fermentation, it is necessary to monitor and control the fermentation during the fermentation process. Typically, the fermentation process is monitored by measuring the fermentation process component concentration (FCC) during the fermentation process.

Traditional methods for measuring FCC include chemical titration, high performance liquid chromatography, and the like. These traditional methods all extract a large amount of fermentation broth for off-line measurement, so it is not possible to measure the crusting in real time and apply it to the control of the fermentation process. In addition, since these conventional methods require a large amount of fermentation broth to be extracted from the fermenter, these methods have a very adverse effect on the fermentation process.

In view of the shortcomings of the above conventional methods, techniques for measuring FCC online have been developed. For example, European Patent Application EP0445675 discloses a flow splitting device based on flow injection analysis (FIA) technology. However, this patent application requires multiple peristaltic pumps and multiple valves when implemented in an FIA system, resulting in a very complicated structure of the flow analysis device, increasing the operational complexity of the entire analysis process, and also resulting in the flow analysis device. The working condition is unstable. In addition, FIA systems are not flexible enough to be used in a variety of chemical reactions.

Based on the FIA, a sequential injection analysis (SIA) technique was proposed in the 1990s. A flow analysis device for in-line titration based on FIA technology is also disclosed in U.S. Patent No. 6,096,274. The apparatus includes a pump, a multi-channel valve, a storage member, a beaker for use as a reaction vessel, a mass sensor, and a conduit connecting them. During the measurement process, the pump is first sucked into the storage member through the multi-channel valve, then pushed into the beaker through the multi-channel valve, then the reagent is taken through the multi-channel valve and filled with the multi-channel valve, and then the reagent is passed through the multi-channel valve. Pushed into the beaker, samples and reagents are in The beaker is stirred for reaction and the measuring sensor is measured in the beaker. Compared to the solution in EP ^{04 4} 5675, this solution provides a simple structure that can be used in a variety of applications. However, the reaction vessel used in this protocol is a beaker, so a large amount of sample and reagent is required for each measurement. On the other hand, it is difficult to thoroughly clean the on-line measurement using the flow analysis device, and residual samples or reagents may affect the next measurement. Summary of the invention

In view of this, it is an object of the present invention to provide an apparatus for measuring the concentration of components in a fermentation process. Another object of the present invention is to provide a method of measuring the concentration of components in a fermentation process.

Accordingly, the present invention provides an apparatus for measuring the concentration of a component of a fermentation process, the apparatus comprising:

a sample flow path for flowing a sample of the driven fermentation broth therein; a reagent flow path for flowing the driven reagent therein; the shape of the reagent flow path being designed such that Forming a cavity along the sample flow path at least partially overlapping the sample flow path and the reagent flow path;

a dialysis membrane positioned between the sample flow path and the reagent flow path to allow analytes in the fermentation broth sample to diffuse from the sample flow path into the reagent flow path for the analyte to The reagent reacts;

A monitoring chamber is disposed on the reagent flow path for measuring the light absorbance of the reagent after the reaction.

In the above technical solution, the sample flow path is a groove formed on an upper part, the reagent flow path is a groove formed on a lower part, and the upper part and/or the lower part is made of poly Made of decyl acrylate (PMMA), glass or silicon.

Alternatively, the sample flow path and the reagent flow path may each be a half tube which is cut in the longitudinal direction.

Preferably, the sample flow path and the reagent flow path are twisted and twisted to extend the flow path in a limited space.

A downstream end of the sample flow path is provided with a fermentation liquid outlet, and a downstream end of the reagent flow path is provided with a reagent outlet. The upper member and the lower member are respectively provided with positioning mechanisms that cooperate with each other, and when the upper member and the lower member are closed, the sample flow path and the reagent flow path are aligned with each other to form a cavity. .

A temperature measuring element and a temperature control element are disposed in the sample flow path, and the temperature control element is used to adjust the temperature of the fermentation liquid sample in the sample flow path.

Preferably, the dialysis membrane may fill the area where the sample flow path is located.

The monitoring chamber is located downstream of the reagent flow path and outside the portion where the reagent flow path overlaps the sample flow path.

Further, one side of the monitoring room is connected to one input fiber, and the other side is connected with an output fiber, the input fiber and the output fiber are aligned with each other; the input fiber is connected to a light source, and the output fiber is Connected to a detector.

Preferably, the optical path length between the input fiber and the output fiber of the monitoring chamber is adjustable.

Further, two slots are disposed in the lower member for respectively fixing the input fiber and the output fiber. '

To incorporate the SIA system, the device is coupled to a sequential injection analysis flow system comprising: a pump, a multi-channel valve, and a reservoir, wherein the pump is coupled to one end of the reservoir The multi-channel valve has a plurality of ports respectively connected to the other end of the liquid storage tube, the inlet of the sample flow path, the inlet of the reagent flow path, the outlet of the reagent container, and an outlet of the container of the fermentation liquid to be measured.

The multi-channel valve further includes: a port connected to the container of the calibration solution and/or a port for discharging the waste liquid.

The invention also proposes a method for measuring the concentration of components in a fermentation process, comprising: inputting a sample of the fermentation broth into a sample flow path, inputting the reagent into a reagent flow path; utilizing between the sample flow path and the reagent flow path Dialyzing the membrane, passing the analyte in the fermentation broth sample through the dialysis membrane into the reagent flow path, and reacting with the reagent; passing light through the post-reaction reagent in the reagent flow path, and measuring the The light absorbance of the reagent after the reaction;

The analyte concentration in the fermentation broth sample is calculated based on the light absorbance.

The method further comprises: adjusting before the measuring the light absorbance of the reagent after the reaction The length of the optical path in the reagent after the reaction.

The method further comprises: replacing the fermentation broth sample with a calibration solution, and measuring an analyte concentration in the calibration solution; analyzing the standard value and the measured concentration of the analyte concentration in the calibration solution The concentration of the analyte, the concentration of the analyte in the sample of the fermentation broth is calibrated.

The method further includes: introducing a cleaning fluid into the sample flow path and/or the reagent flow path to clean the sample flow path and/or the reagent flow path.

As can be seen from the above scheme, since the device of the present invention adopts a unique structure, it is not necessary to use a large-capacity container for the reaction, and only a small amount of the fermentation broth sample and the reagent can be used for the measurement. , reducing the adverse effects on the fermentation process. The device of the present invention can be directly rinsed with a cleaning solution, and the cleaning process is simple, and the fermentation liquid sample and reagent are not easily left, thereby avoiding the problem that the residual fermentation liquid sample or reagent affects the next measurement. The sensitivity of the measurement can be varied by adjusting the length of the light path in the reagent after the reaction. With the technical solution of the present invention, it is not necessary to perform a pretreatment step of separating the fermentation broth sample and the reagent, thereby simplifying the measurement step. DRAWINGS

1 is an exploded perspective view of an apparatus for online measurement of a component concentration in a fermentation process according to an embodiment of the present invention;

Figure 2 is a schematic view of the device shown in Figure 1 as a whole;

Figure 3 is a schematic view of the upper part when the apparatus of Figure 1 further includes a temperature control element and a temperature measuring element;

Figure 4 is a schematic view showing the structure of an SIA system connected to the apparatus shown in Figure 1. detailed description

In order to make the objects, technical solutions and advantages of the present invention more comprehensible, the present invention will be further described in detail below.

The technical solution proposed by the present invention can be used to measure the concentration of components in the fermentation process during fermentation, such as the concentration of the substrate or the concentration of the product. Since glucose is an important carbon source in many fermentation processes, glucose is used as the analyte in the following description. The solution of the present invention is exemplified, but the present invention is not limited to measuring the glucose concentration, and other analytes such as glutamic acid, lactic acid, ethanol and the like can also be measured by selecting the corresponding reagent.

The detection methods of analytes such as glucose can be roughly classified into two types, one is an enzyme-based electrochemical (EC) method, and the other is a photometric method. The present invention mainly relates to photometric methods.

Since the light absorbance of the solution after the reaction of the glucose and the reagent is proportional to the concentration of glucose, the photometric method obtains the concentration of glucose by measuring the light absorbance of the solution after the reaction of the glucose and the reagent. The photometric method can be very accurate where the temperature and pH of the solution remain stable. Also, at room temperature, a typical measurement can be done quickly in less than a minute, allowing measurements to be used to control the fermentation process in real time.

As shown in the chemical formulas (1) and (2), glucose is oxidized by glucose oxidase to form gluconic acid and hydrogen peroxide, while hydrogen peroxide is reacted with peroxylase and 4-amino-anthracene. Bilin (4-APP) and N-ethyl-N-sulfopropyl-m-mercaptoaniline (TOPS) react to form quinone. The quinone imine has a color and has an absorption peak for light having a wavelength of 545 nm.

Glucose + H ₂ O + 0 ₂ —— > Gluconic acid + H ₂ 0 ₂ ( 1 )

2 H ₂ 0 ₂ + 4-AAP + TOPS —— > S ketimine + 4 H ₂ 0 ( 2 ) Since the reagent solution has an absorption peak at 545 nm after the reaction, and the absorption of the reagent solution for light after the reaction The concentration of glucose in the fermentation broth is directly proportional, and thus embodiments of the present invention can utilize this feature to measure the concentration of glucose in the fermentation broth. The relationship between light absorbance and glucose concentration follows the Lambert-Beer law as shown in equation (3).

A = sLC ( 3 ) where A represents the light absorbance, the coefficient represents the light absorbance of the solution per unit mole (mol), L is the optical path length in the solution, and C is the concentration of the analyte, in this embodiment C For the glucose concentration. According to formula (3), when and L are determined, by measuring A, the glucose concentration can be calculated (:.

Based on the above principle, an embodiment of the present invention provides a device for measuring the concentration of components in a fermentation process on-line, the device measuring the light absorbance of the reagent solution after the reaction, The concentration of glucose in the fermentation broth is obtained, so that the state of fermentation can be known.

As shown in FIG. 1, the apparatus 100 for measuring the concentration of components in a fermentation process includes an upper member 101, a lower member 102, a dialysis membrane 103, an input fiber 106, an output fiber 107, a light source 108, and a detector 109. .

The upper member 101 includes a sample flow path 104 through which the fermentation broth sample flows. The lower member 102 includes a reagent flow path 105 through which the reagent flows. The upper member 101 and the lower member 102 are joined together such that the sample flow path 104 and the reagent flow path 105 at least partially overlap each other, thereby forming a cavity along the sample flow path 105. For example, the sample flow path may be a groove formed in the upper member 101, and the reagent flow path 105 may be a groove formed in the lower member 102. In this case, the upper member 101 and the lower member 102 may be respectively provided with positioning mechanisms that cooperate with each other such that when the upper member 101 and the lower member 102 are closed, the sample flow path 104 and the reagent flow path 105 are overlapped with each other. Form a cavity. The upper member 101 and/or the lower member 102 may be made of a material such as polymethyl methacrylate (PMMA), glass or silicon.

Additionally, the device 100 can be constructed in a more compact configuration. For example, the sample flow path 104 and the reagent flow path 105 are respectively a half pipe which is longitudinally cut, thereby eliminating the upper member 101 and the lower member 102.

Further, the sample flow path 105 and the reagent flow path 105 may be arranged in a meandering manner, for example, arranged in a "Z" shape, an "S" shape, a sinusoidal function shape, or a shape as shown in FIG. 1, thereby extending the flow path in a limited space. In order to facilitate the reaction of the analyte and the reagent.

The dialysis membrane 103 is located at least between the sample flow path 104 and the reagent flow path 105, and the area of the dialysis membrane 103 may be larger than a portion where the fermentation sample flow path 104 and the reagent flow path 105 overlap, for example, the area where the sample flow path 104 is filled. It is even possible to cover the entire portion where the upper part 101 and the lower part 102 are in contact, which facilitates assembly. The dialysis membrane 103 separates the sample flow path 104 from the reagent flow path 105 to form two independent flow paths. The width of the two flow paths can be tens of microns to several centimeters.

The sample flow path 104 has a sample inlet 110 and a sample outlet 11 1, the sample inlet 1 10 for the fermentation liquid sample to flow in, and the sample outlet 1 11 for the fermentation liquid sample to flow out. The reagent flow path 105 has a reagent inlet 1 12 and a reagent outlet 1 13, and a reagent inlet 112 for The reagent flows in, and the reagent outlet 113 supplies the reagent out. In addition, the dialysis membrane 103 located between the sample flow path 104 and the reagent flow path 105 can allow analytes such as glucose in the sample of the fermentation liquid in the sample flow path 104, and small molecules such as salt ions to pass, that is, glucose is allowed to be The analyte diffuses from the fermentation broth sample into the reagent of the reagent flow path 105, and those large or large particles having a diameter larger than the pore diameter of the dialysis membrane 103 are blocked by the dialysis membrane 103.

The dialysis membrane 103 may be a cellulose membrane, a polycarbonate membrane or the like. The pore diameter of the dialysis membrane 103 needs to be larger than the diameter of the analyte such as glucose. Preferably, the diameter of the pores of the dialysis membrane 103 can be less than 0.22 μm, so that it is possible to block bacteria and the like while transmitting an analyte such as glucose. Since the dialysis membrane 103 separates the fermentation broth sample and the reagents and allows the glucose to permeate and blocks the large particles, the centrifugation of the fermentation broth sample and the reagent pretreatment are not required before the measurement, thereby shortening the measurement time. If it is used for a long period of time, it is possible to reduce the dialysis efficiency of the dialysis membrane 103. The apparatus 100 of the embodiment of the present invention can inject a calibration solution into the sample flow path and obtain a calibration solution measurement result, and use the measurement result and calibration. The standard value of the solution can be used to calibrate the measurement of the actual fermentation broth sample.

A monitoring chamber 114 is disposed in the reagent flow path 105. The monitoring chamber 1 14 can be located anywhere in the reagent flow path 105, preferably downstream of the reagent flow path 105, to ensure that there is a fully reacted post-reaction reagent in the monitoring chamber 114. Alternatively, the monitoring chamber 114 may be located within the portion of the reagent flow path 105 that is aligned with the sample flow path 104, or may be located outside of the portion of the reagent flow path 105 that is aligned with the sample flow path 104. In the example shown in the drawings, the monitoring chamber 114 is located outside the portion of the reagent flow path 105 that is aligned with the sample flow path 104.

The monitoring chamber 114 is connected to an input fiber 106 and an output fiber 107. The length L of the optical path in the monitoring chamber 114, i.e., the distance between the input fiber 106 and the output fiber 107, can be designed to be adjustable. It can be seen from the formula (3) that by adjusting the optical path length of the monitoring chamber 144, the sensitivity of the measurement can be adjusted very easily, and thus applied to different fermentation broth samples. The input fiber 106 and the output fiber 107 may be misaligned or aligned. In the case of misalignment, the output optical fiber 107 receives less light intensity than that received under alignment, so the present invention preferably employs two The structure of the alignment. Two aligned or misaligned guiding grooves can be formed at both ends of the monitoring chamber 114 in the lower member 102, and the input optical fiber 106 and the output optical fiber 107 can be respectively fixed in the guiding groove, so that it can be both accurate and convenient. The input fiber 106 and the output fiber 107 are fixed. The incident light input from the input fiber 106 passes through the post-reaction reagent solution in the monitoring chamber 114 and is received by the output fiber 107. The light absorbance of the reagent solution after the reaction can be calculated based on the input light intensity and the output light intensity, and then the glucose concentration in the fermentation liquid can be calculated according to the formula (3).

The light source 108 is used to generate incident light. A laser diode, a light emitting diode or the like can be used. The output spectral range of the light source 108 includes an absorption peak of an analyte such as glucose. It should be noted that the incident light generated by the light source 108 is not necessarily a laser, but may be other types of light such as white light. The detector 109 can be a photodiode, a photomultiplier tube or the like for measuring the intensity of light passing through the reagent solution after the reaction. The light absorbance A in the formula (3) can be calculated by using the intensity of the light output from the light source 108 and the intensity of the light measured by the detector 109.

The combination of the structures shown in Fig. 1 can form a device 100 as shown in Fig. 2, which may be referred to as a small on-line photometer (MOP) integrated chip. The apparatus 100 can be fabricated using a fine processing or microelectromechanical system (MEMS) processing process, and the material can be made of polymethyl methacrylate, glass or silicon or the like according to various needs.

The process of measuring the concentration of components in the fermentation process using the apparatus shown in Figures 1 and 2 can include the following steps:

Step S1 1 , a sample of the fermentation broth is extracted from a fermentation broth container such as a fermenter, and a sample of the fermentation broth is introduced into the sample flow path 104 through the sample inlet 110.

In step S12, the reagent is introduced into the reagent flow path 105 through the reagent inlet 112.

The above steps S1 1 and S12 do not have a strict sequence, and may be performed simultaneously, or step S11 or S 12 may be performed first.

In step S13, since the concentration of the analyte in the fermentation liquid sample in the sample flow path 104 and the reagent flow path 105 is different, the analyte in the fermentation liquid sample passes through the pore of the dialysis membrane 103 into the reagent. And chemically reacting with related substances in the reagent, for example, the reactions shown by the chemical formulas (1) and (2), forming a post-reaction reagent including the reaction product, and the reagent is colored after the reaction.

Step S14, it is possible to wait for a period of time so that the reaction proceeds sufficiently. Then, light is output by the light source 108, and the light enters the monitoring chamber 113 through the input fiber 106. After absorption in the monitoring chamber 113, the reagent is absorbed by the output fiber 107 and transmitted to the detector 109. In step S15, the detector 109 measures the intensity of the light absorbed by the reagent after the reaction. Step S16, the light absorbance A of the reagent after the reaction is calculated based on the intensity of the light output from the light source 108 and the intensity of the light measured by the detector 109.

Step S17, based on the calculated light absorbance and the optical path length L in the coefficient solution, the concentration C of the analyte is calculated according to the formula (1), so that the fermentation state can be known.

Step S18, the fermentation broth sample is output from the sample outlet 1 1 1 , and the reagent is output from the reagent outlet 1 13 .

The method may further include: step S19, inputting the cleaning liquid from the sample inlet 110 and outputting from the sample outlet 111, and inputting the cleaning liquid from the reagent inlet 112 and outputting from the reagent outlet 113, thereby cleaning the device 100 for the next Used for secondary measurements.

The method may also include a calibration process: performing the above steps S11 to S18 with the calibration solution instead of the fermentation liquid, thereby measuring the concentration of the analyte in the calibration solution; using the standard value of the analyte concentration in the calibration solution and The resulting analyte concentration is measured to calibrate the measured concentration in the measured fermentation broth sample.

It can be seen from the formula (3) that the smaller the L, the higher the measured sensitivity. Therefore, for different fermentation broths, the above method can also adjust the optical path length L in the detection chamber 114 to change the sensitivity of the measurement. For example, increasing the length L of the optical path reduces the sensitivity of the measurement; reduces the length L of the optical path to increase the sensitivity of the measurement.

Fig. 3 is another embodiment of the apparatus for online measurement of the concentration of components in a fermentation process in the present invention. The apparatus shown in FIG. 3 adds a temperature control for controlling the temperature of the fermentation broth sample in the sample flow path 104 in the sample flow path 104 of the upper member 101 on the basis of the apparatus 100 shown in FIGS. 1 and 2. A component, such as a Peltier line 201. By applying a current in a different direction to the Peltier line 201, the temperature of the fermentation broth sample in the sample flow path 104 can be controlled. For example, when a current is applied in one direction, the Peltier line 201 absorbs heat, thereby lowering the temperature of the fermentation liquid sample, and when the current is applied in the other direction, that is, in the opposite direction, the Peltier line 201 emits heat, thereby increasing the fermentation liquid. The temperature of the sample.

The apparatus shown in Fig. 3 can further provide a temperature measuring element for measuring the temperature of the fermentation liquid sample in the medium sample flow path 104 in the sample flow path 104 of the upper member 101, such as a temperature sensor 202, a thermometer, or the like. When the above apparatus 100 includes both the temperature control element and the temperature measuring element, closed loop control of the temperature of the fermentation liquid in the sample flow path 104 can be achieved. Taking the Peltier line 201 and the temperature sensor 102 as an example, the temperature of the fermentation broth sample is measured by the temperature sensor 202. When the temperature is too high, the temperature of the fermentation broth sample is lowered by applying a current to the Peltier line 201. When low, the temperature of the fermentation broth sample is increased by applying a current to the Peltier line 201. In this way, the temperature of the fermentation broth sample can be controlled within a suitable range.

Figure 4 shows an embodiment in which a device for measuring the concentration of components in a fermentation process is combined with an SIA flow system 300. The apparatus for measuring the concentration of components in the fermentation process on-line in Fig. 4 may employ the apparatus 100 shown in Fig. 1, Fig. 2 or Fig. 3, and will not be described again.

The SIA flow system 300 shown in Figure 4 includes a pump 301, "- a multi-channel valve 302, a reservoir 303, and a line connecting them. The pump 301 is coupled to one end of the reservoir 303. The reservoir 303 The other end, the inlet 110 of the sample flow path 104, the inlet 112 of the reagent flow path 105, the fermentation liquid container (e.g., fermentor, not shown), and the reagent container 304 are respectively connected to one port of the multi-channel valve 302. The end to which the pump 301 and the liquid storage tube 303 are connected is also connected to a cleaning liquid container 305. Also, the multi-channel valve 302 has a discharge port for discharging waste liquid. The above measurement fermentation can be completed by the SIA flow system 300. The process of process component concentration.

During the measurement, the pump 301 is connected to the fermentation liquid container through the liquid storage tube 303, the multi-channel valve 302, and the sample of the fermentation liquid in the fermentation liquid container is pumped into the liquid storage tube 303 by the pump 301, and then the pump 301 is passed through the storage. The liquid tube 303, the multi-channel valve 302 is connected to the sample inlet 110 of the sample flow path 104, and the fermentation liquid sample in the liquid storage tube 303 is pushed to the sample inlet 110 by the pump 301, thereby inputting the fermentation liquid sample into the sample flow path 104. The reservoir 303 is purged by the pump 301 to purge the reservoir 303, and the purge is discharged from the discharge port of the multi-channel valve 302. The pump 301 is connected to the reagent container 304 through the reservoir 303, the multi-channel valve 302, the reagent in the reagent container 304 is pumped into the reservoir 303 by the pump 301, and then the pump 301 is passed through the reservoir 303, the multi-channel valve 302 is connected to the reagent inlet 112 of the reagent flow path 104, and the reagent in the liquid storage tube 303 is pushed by the pump 301 to the reagent inlet 112 of the reagent flow path 105, thereby inputting the reagent into the reagent flow path 105. By the above operation, the fermentation broth sample and the reagent can be separately input into the sample flow path 104 and the reagent flow path 105 of the apparatus 100. At this time, in the sample flow path 104, the analyte such as glucose in the fermentation liquid sample is diffused into the reagent of the reagent flow path 105 through the dialysis membrane 103, and reacts with the reagent. If you need to, you can wait a while for the reaction to be more adequate. By measuring the post-reaction reagent in the monitoring chamber 114 downstream of the reagent flow path 105, the light absorption rate A is obtained, and then the glucose-glucose concentration can be calculated according to the formula (3), so that the glucose concentration in the fermentation liquid can be known. .

Finally, the cleaning liquid in the cleaning liquid container 305 is pumped by the pump 301, and the cleaning liquid is pushed to the sample flow path 104 and the reagent flow path 105 through the liquid storage tube 303 and the multi-channel valve 302, respectively, to clean the two flow paths. The cleaned liquid can be directly discharged from the sample outlet 111 of the sample flow path 104 and the reagent outlet 113 4 of the reagent flow path 105.

As shown in Figure 4, the multi-channel valve 302 in the SIA flow system 300 can also have a port that is coupled to a calibration solution container 306 containing a calibration solution. The concentration of the analyte in the calibration solution is measured by the above process, and the analyte concentration of the measured fermentation broth sample can be calibrated using the measured analyte concentration and the actual analyte concentration of the calibration solution.

The above is only the preferred embodiment of the present invention, and is not intended to limit the present invention. Any modifications, equivalents, improvements, etc., which are included in the spirit and scope of the present invention, should be included in the present invention. Within the scope of protection.

Claims

1. A device (100) for measuring the concentration of a component of a fermentation process, the device (100) comprising:

a sample flow path (104) for flowing a sample of the driven fermentation broth therein; a reagent flow path (105) for flowing a driven reagent therein; said reagent flow path (105) The shape is such that the sample flow path (104) and the reagent flow path (105) at least partially overlap each other to form a cavity along the sample flow path (105);

a dialysis membrane (103) located between the sample flow path (104) and the reagent flow path (105), allowing analytes in the fermentation broth sample to diffuse from the sample flow path (104) to the a reagent flow path (105) for reacting the analyte with the reagent; a monitoring chamber (114) disposed on the reagent flow path for measuring the light absorbance of the reagent after the reaction.

2. Apparatus (100) according to claim 1, wherein said sample flow path (104) is a groove formed in an upper part (101), said reagent flow path (105) being formed in a lower portion A groove on the component (102), the upper component (101) and/or the lower component (102) is made of polymethyl methacrylate (PMMA), glass or silicon material.

The apparatus (100) according to claim 1, wherein the sample flow path (104) and the reagent flow path (105) are each a half tube cut longitudinally.

The apparatus (100) according to any one of claims 1 to 3, wherein the sample flow path (104) and the reagent flow path (105) are arranged in a meandering manner to extend the flow path in a limited space.

The device (100) according to any one of claims 1 to 3, wherein a downstream end of the sample flow path (104) is provided with a fermentation liquid outlet, and a downstream end of the reagent flow path (105) is provided. There is a reagent outlet.

The device (100) according to claim 2, wherein the upper member (101) and the lower member (102) are respectively provided with positioning mechanisms that cooperate with each other, the upper member and the lower member When closed, the sample flow path (104) and the reagent flow path (105) are aligned with each other to form a cavity. The device (100) according to any one of claims 1 to 3, wherein a temperature measuring element (202) and a temperature control element (201) are disposed in the sample flow path (104), the temperature A control element (201) is used to adjust the temperature of the fermentation broth sample in the sample flow path (104).

8. Apparatus (100) according to claim 1, wherein said dialysis membrane (103) can fill the area of said sample flow path (104).

9. The apparatus (100) according to claim 1, wherein the monitoring chamber (114) is located downstream of the reagent flow path (105) and is located in the reagent flow path (105) and the sample flow Road (104) is outside the overlap.

10. Apparatus (100) according to claim 1 or 9, wherein said monitoring room

An input fiber (106) is connected to one side of (114), and an output fiber (107) is connected to the other side, and the input fiber (106) and the output fiber (107) are aligned with each other; (106) is coupled to a light source (108) that is coupled to a detector (109).

11. Apparatus (100) according to claim 1, wherein the length of the optical path between the input fiber (106) and the output fiber (107) of the monitoring chamber (114) is adjustable.

12. Apparatus (100) according to claim 10, wherein said lower part (102) is provided with two slots for securing said input fiber (106) and output fiber (107), respectively.

13. Apparatus (100) according to claim 1, wherein the apparatus (100) is coupled to a sequential injection analysis flow system (300), the sequential injection flow system

(300) comprising: a pump (301), a multi-channel valve (302) and a liquid storage tube (303), wherein the pump (301) is connected to one end of the liquid storage tube (303), the multi-channel The valve (302) has a plurality of ports respectively connected to the other end of the liquid storage tube (303), the inlet of the sample flow path (104), the inlet of the reagent flow path (105), the outlet of the reagent container (304), and An outlet of the container of the fermentation broth to be measured.

14. Apparatus (100) according to claim 13, wherein said multi-channel valve (302) further comprises: a port connected to the container of the calibration solution and/or a port for discharging the waste liquid.

15. A method of measuring the concentration of a component of a fermentation process, comprising:

Enter the fermentation broth sample into a sample flow path ( 104) and enter the reagent into a reagent stream Road (105);

Using the dialysis membrane (103) between the sample flow path and the reagent flow path, the analyte in the fermentation broth sample is passed through the dialysis membrane (103) into the reagent flow path (105), and Reagent reaction;

Passing light through the post-reaction reagent in the reagent flow path (105), and measuring the light absorbance of the reagent after the reaction;

16. The method according to claim 15, wherein the method further comprises: adjusting the optical path length in the reagent after the reaction before the measuring the light absorbance of the reagent after the reaction.

17. The method according to claim 15, wherein the method further comprises: replacing the fermentation broth sample with a calibration solution, and measuring an analyte concentration in the calibration solution;

The analyzed concentration in the fermentation broth sample is calibrated based on the standard value of the analyte concentration in the calibration solution and the measured analyte concentration.