Information delivery methods, analysis systems, and programs

Abstract

According to the present invention, an information processing device acquires (step S10) the weight of a metabolite identified in a liquid culture in a container, and acquires (step S20) the weight of a microorganism identified in the liquid culture in the container. In addition, the information processing device derives (step S30), by using the inverse number of the weight of the microorganism and the weight of the metabolite, an index concerning the weight of the metabolite, and outputs (step S40) the index.

Description

Technical Field

[0001] The present disclosure relates to a method for providing information on metabolites, an analysis system, and a program.

Background Art

[0002] As disclosed in Patent Document 1, an apparatus for culturing cells such as microorganisms by adjusting the dissolved oxygen concentration while stirring a culture solution in a container is known.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] The weight of metabolites of a microorganism may be used for the analysis of the microorganism. In such a case, there has been a need for a technique for a researcher to handle the weight of the metabolite in consideration of the environment in which the metabolite was produced.

[0005] The present disclosure has been made to solve such problems, and an object thereof is to provide a technique for handling the weight of a metabolite in a state where the environment in which the metabolite was produced is taken into consideration.

Means for Solving the Problems

[0006] A first aspect of the present disclosure is a method for providing information on metabolites, including steps of obtaining the weight of a specific metabolite in a culture solution in a container, obtaining the weight of a specific microorganism in the culture solution in the container, deriving an index regarding the weight of the metabolite using the weight of the metabolite and the reciprocal of the weight of the microorganism, and outputting the index.

[0007] A second aspect of the present disclosure relates to an analytical system for providing information on the weight of microbial metabolites, comprising an information processing device, the information processing device being configured to obtain the weight of identified metabolites in a microbial culture medium in a container, obtain the weight of identified microorganisms in the culture medium, derive an index relating to the weight of metabolites using the weight of the metabolites and the reciprocal of the weight of the microorganisms, and output the index.

[0008] A third aspect of the present disclosure relates to a program executed by a computer that provides information on metabolites, which causes the computer to perform the steps of: obtaining the weight of a metabolite identified in a culture medium in a container; obtaining the weight of a microorganism identified in a culture medium in a container; deriving an index relating to the weight of a metabolite using the reciprocal of the weight of the metabolite and the weight of the microorganism; and outputting the index. [Effects of the Invention]

[0009] This disclosure provides information for handling the weight of metabolites in a manner that takes into account the environment in which the metabolites were produced. [Brief explanation of the drawing]

[0010] [Figure 1] This is a diagram showing the schematic configuration of the automated preprocessing system. [Figure 2] This is a flow path diagram showing the flow path configuration of the sampling device. [Figure 3] This is a block diagram illustrating the schematic configuration of the control device. [Figure 4] This figure shows the skim for obtaining the weight of metabolites and the weight of microorganisms in the culture medium of cell culture device 100. [Figure 5] This is a flowchart for the process of outputting an index of metabolite weights. [Figure 6]This figure shows the volume of liquid supplied to the tubes for gravimetric analysis of microorganisms and the volume of liquid supplied to the tubes for gravimetric analysis of metabolites in the analysis system 700. [Modes for carrying out the invention]

[0011] This embodiment will be described in detail with reference to the drawings. Note that identical or corresponding parts in the drawings are denoted by the same reference numerals, and their descriptions will not be repeated in principle. <Outline of the analysis system configuration> Figure 1 is a block diagram illustrating the schematic configuration of the analysis system 700. The analysis system 700 includes a liquid chromatograph-mass spectrometer 3, an automated sample preparation system 10, and an information processing device 70. The automated sample preparation system 10 is a device for automatically performing sample preparation on the analyte for the liquid chromatograph-mass spectrometer 3. An example of an analyte is cultured microorganisms. Another example is microbial metabolites. The information processing device 70 can communicate with the automated sample preparation system 10 and the liquid chromatograph-mass spectrometer 3 and controls the analysis system 700 as a whole.

[0012] The information processing device 70 includes a CPU (Central Processing Unit) 51, memory 72, communication interface 73, input device 74, and display 75. The memory 72 is composed of, for example, ROM (Read Only Memory) and RAM (Random Access Memory), and can store control programs as well as various other data non-temporarily.

[0013] The communication interface 73 is implemented, for example, by a network card, and enables the information processing device 70 to communicate with the automated preprocessing system 10 and the liquid chromatograph mass spectrometer 3. The input device 74 inputs data from the user to the CPU 71 and is implemented, for example, by a keyboard and / or mouse. The display 75 displays the calculation results of the CPU 71 and is implemented, for example, by a liquid crystal display.

[0014] The automatic pretreatment system 10 includes a sampling device 1 and a pretreatment device 2. In the automatic pretreatment system 10, metabolites of the microorganism are extracted from the microorganism after pretreatment. The extracted metabolites are supplied to a liquid chromatograph mass spectrometer 3. The liquid chromatograph mass spectrometer 3 is merely an example of an analyzer for analyzing an analyte. It is also possible to analyze the analyte using other analyzers.

[0015] The sampling device 1 is a device for sampling liquid from a container (culture container). For example, microorganisms are cultured in a culture solution containing a culture medium in a container called a bioreactor. In the bioreactor, for example, a stirring member rotated using magnetic force, an oxygen concentration sensor for detecting the concentration of dissolved oxygen, etc. are provided. By adjusting the dissolved oxygen concentration while stirring the culture solution containing the culture medium and microorganisms in the bioreactor, the microorganisms are cultured in the sampling device 1.

[0016] The pretreatment device 2 performs pretreatment on the microorganisms contained in the culture solution (culture sample) sampled from inside the bioreactor. In the sampling device 1, the culture solution is accommodated in a tube. The pretreatment device 2 includes a centrifugation mechanism 4, a liquid removal mechanism 5, a reagent supply mechanism 6, a stirring mechanism 7, and an extraction mechanism 8. These respective mechanisms sequentially perform pretreatment on the culture solution in the tube.

[0017] The centrifugation mechanism 4 applies centrifugal force to the culture solution in the tube. As a result, the culture solution in the tube separates into a solid component that sinks to the bottom of the tube with the solid-liquid interface as the boundary, and a liquid component that floats on top of the solid component. The solid component is the culture (for example, the cultured microorganisms). The liquid component that floats on top of the solid component is the supernatant separated from the culture solution.

[0018] The liquid removal mechanism 5 sucks the supernatant from the tube. As a result, the liquid in the tube is removed, and the microorganisms remain in the tube.

[0019] The reagent supply mechanism 6 supplies a reagent for extracting metabolites of microorganisms into the tube. Thereby, a mixed solution of microorganisms and the reagent is generated in the tube.

[0020] The stirring mechanism 7 stirs the mixed solution. By stirring the mixed solution, a suspension containing metabolites of microorganisms can be obtained.

[0021] The extraction mechanism 8 extracts a part of the suspension as an extract. The extract is supplied to the liquid chromatograph mass spectrometer 3.

[0022] <Schematic configuration of the sampling device> FIG. 2 is a flow path diagram showing the flow path configuration of the sampling device 1. In the sampling device 1, a culture solution containing microorganisms in a cell culture device 100 called a bioreactor is sampled. The cell culture device 100 includes a stirrer 111 as a stirring member rotated using magnetic force inside it.

[0023] The cell culture device 100 is held by a holding part 12 provided in the sampling device 1. In the present embodiment, three cell culture devices 100 can be held by one holding part 12, and a plurality (for example, four) of such holding parts 12 are provided. The holding part 12 may be configured to be provided only one. The holding part 12 may also be configured to hold two or less or four or more cell culture devices 100.

[0024] The cell culture device 100 can perform culturing in a heated state by a heater (not shown) provided in the holding part 12. A motor 13 for rotating a magnet (not shown) is connected to the holding part 12. By rotating this motor 13, the magnet can be rotated, and the stirrer 111 in each cell culture device 100 can be rotated by the magnetic force.

[0025] The sampling device 1 can perform cell culture by controlling the temperature of the culture medium in the cell culture apparatus 100 and stirring the culture medium with the stirring bar 111. The sampling device 1 samples the culture medium containing the cultured microorganisms into the tube 14 at any desired timing.

[0026] The sampling device 1 is equipped with a culture medium sampling mechanism 20 for sampling the culture medium into a tube 14, and a reagent sampling mechanism 30 for sampling the reagent into a tube 14. An example of a reagent is a reagent (quencher) for stopping the metabolic reaction of microorganisms in the culture medium. The tube 14 contains a mixture of the culture medium and the reagent, is sealed with a cap (not shown), and then transported to the pretreatment device 2. Alternatively, a tube 14 containing only the culture medium (without mixing with the reagent) may be transported to the pretreatment device 2.

[0027] The culture medium sampling mechanism 20 is equipped with a pump 21 and a number of valves 22 and 23. Each valve 23 has, for example, one pair of common ports and five pairs (10 in total) of selective ports. The flow path can be switched by arbitrarily selecting one pair of selective ports and connecting it to one pair of common ports.

[0028] The pump 21 and valve 22 are installed in a flow path 41 that connects a pair of common ports. The valve 22 constitutes a flow path switching unit (first flow path switching unit) for switching whether or not to guide the liquid in the flow path 41 to a branch flow path 42 that branches off from the flow path 41. In other words, the valve 22 can switch between a state in which liquid flows between the pair of common ports via the flow path 41, or a state in which the liquid in the flow path 41 is guided to the branch flow path 42.

[0029] Of the five pairs of selection ports, one pair is connected to an outlet 43 and an inlet 44, respectively, which communicate with one cell culture device 100. The outlet 43 is a channel for draining the culture medium from within the cell culture device 100. The inlet 44, on the other hand, is a channel for introducing the culture medium, which is drained from the cell culture device 100 via the outlet 43 and circulated via the channel 41, back into the cell culture device 100. Another pair of selection ports is connected to an outlet 45 and an inlet 46, respectively, which communicate with another cell culture device 100. Yet another pair of selection ports is connected to an outlet 47 and an inlet 48, respectively, which communicate with yet another cell culture device 100.

[0030] In the sampling device 1, any of the outlet passages 43, 45, or 47 and their corresponding inlet passages 44, 46, or 48 are connected via the flow path 41, and the pump 21 is driven in this state, thereby circulating the culture medium within each cell culture device 100. In other words, the flow path 41, each outlet passage 43, 45, or 47, and each inlet passage 44, 46, or 48 constitute a circulation flow path (first circulation flow path) for circulating the culture medium within each cell culture device 100.

[0031] The pump 21 discharges the culture medium from each cell culture device 100 into the first circulation channel and introduces the culture medium into each cell culture device 100 from the first circulation channel, thereby constituting a circulation mechanism (first circulation mechanism) that circulates the culture medium within each cell culture device 100 via the first circulation channel.

[0032] Each outlet 43, 45, and 47 has its tip immersed in the culture medium within the corresponding cell culture device 100. On the other hand, each inlet 44, 46, and 48 has its tip positioned above and spaced apart from the culture medium within the corresponding cell culture device 100. The culture medium discharged from the cell culture device 100 via each outlet 43, 45, and 47 and circulating through the channel 41 is introduced into the cell culture device 100 by falling from the tips of each inlet 44, 46, and 48.

[0033] In the sampling device 1, at least the portion of the flow path 41 connecting a pair of common ports that is equipped with a pump 21 is made of a flexible tube. The pump 21 is, for example, a tubing pump, which can pump the liquid inside the tube by deforming (compressing and relaxing) the flexible tube.

[0034] By switching the valve 22, which serves as a first channel switching unit located in the middle of channel 41, the culture medium circulating into each cell culture device 100 via channel 41 can be discharged into branch channel 42. At this time, the tip of branch channel 42 is located inside tube 14, and the culture medium is sampled into tube 14 via branch channel 42.

[0035] Of the two pairs of selection ports other than the three pairs to which each of the outlets 43, 45, 47 and each of the inlets 44, 46, 48 are connected, one pair of selection ports is connected to the cleaning solution tank 26 and the waste liquid tank 27, respectively. The remaining pair of selection ports is connected to the filter 25 and the waste liquid tank 27, respectively. The cleaning solution tank 26 contains a cleaning solution for cleaning the flow path of the culture medium.

[0036] After sampling the culture medium from one of the cell culture devices 100 into tube 14, the valve 23 is switched to connect the washing solution tank 26 and the waste liquid tank 27 to the flow path 41. When the pump 21 is then driven, the washing solution in the washing solution tank 26 is discharged into the waste liquid tank 27 via the flow path 41. This allows the flow path 41 and valve 22, etc., to be cleaned with the washing solution.

[0037] After cleaning with the cleaning solution, the valve 23 is switched to connect the filter 25 and the waste liquid tank 27 to the flow path 41. When the pump 21 is then driven, air is introduced into the flow path 41 via the filter 25 and discharged into the waste liquid tank 27 along with any remaining moisture in the flow path 41. This removes moisture from the flow path 41 and the valve 22, etc.

[0038] The reagent sampling mechanism 30 is equipped with a pump 31 and several valves 32, 33. Each valve 33 has, for example, one common port and several selective ports, and the flow path can be switched by arbitrarily selecting one of the selective ports and connecting it to the common port.

[0039] Pump 31 and valve 32 are installed in a flow path 49 that communicates with the reagent tank 34 at both ends. The reagent tank 34 contains reagents to be mixed with the culture medium sampled in the tube 14. Flow path 49 constitutes a circulation flow path (second circulation flow path) for circulating the reagents in the reagent tank 34. Pump 31 discharges reagents from the reagent tank 34 into the second circulation flow path and introduces reagents into the reagent tank 34 from the second circulation flow path, thereby constituting a circulation mechanism (second circulation mechanism) for circulating the reagents in the reagent tank 34 via the second circulation flow path.

[0040] In the sampling device 1, at least the portion of the flow path 49, which is connected at both ends to the reagent tank 34, that contains the pump 31 is made of a flexible tube. The pump 31 is, for example, a tubing pump, and by deforming (compressing and relaxing) the flexible tube, it can deliver the reagent inside the tube.

[0041] Valve 32 constitutes a flow path switching unit (second flow path switching unit) for switching whether or not to direct the liquid in flow path 49 to branch flow path 50 which branches off from flow path 49. In other words, valve 32 can switch between a state in which the reagent in reagent tank 34 is circulated via flow path 49, or a state in which the reagent in flow path 49 is directed to branch flow path 50.

[0042] In this way, by switching the valve 32, which acts as a second flow path switching unit located in the middle of the flow path 49, the reagent circulating into the reagent tank 34 via the flow path 49 can be made to flow out into the branched flow path 50. The branched flow path 50 is connected to the common port of the valve 33, and one of the selectable ports of the valve 33 is connected to the inside of the tube 14. Therefore, by connecting the selectable port connected to the inside of the tube 14 to the common port, the reagent flowing out from the flow path 49 into the branched flow path 50 can be sampled into the tube 14.

[0043] <Outline configuration of the control device> Figure 3 is a block diagram showing the schematic configuration of the control device 60. The sampling device 1 is equipped with the control device 60. The control device 60 includes, for example, a CPU 61 and a memory 62. The memory 62 is composed of, for example, ROM and RAM, and can store various data non-temporarily in addition to control programs. The CPU 61 can control the operation of the motor 13, pumps 21, 31 and valves 22, 23, 32, 33, etc., by executing the control programs stored in the memory 62.

[0044] The control device 60 can circulate the culture medium in any of the cell culture devices 100 by driving the pump 21 at a constant liquid delivery rate while any of the outlet passages 43, 45, 47 and the corresponding inlet passages 44, 46, 48 are connected via the flow path 41. Based on the control program, the control device 60 can switch the valve 22 for a predetermined time, connecting the flow path 41 and the branched flow path 42, thereby sampling the culture medium in the flow path 41 into the tube 14.

[0045] The control device 60 can control the amount of culture medium sampled by controlling the time it takes to switch the flow path with the valve 22. In other words, if the pumping speed of the pump 21 is known in advance, the desired amount of culture medium can be accurately sampled into the tube 14 by adjusting the time it takes to connect the flow path 41 and the branched flow path 42.

[0046] The control device 60 can circulate the reagents in the reagent tank 34 by driving the pump 31 at a constant liquid delivery rate while the flow path 49 is connected from one end to the other. Based on the control program, the control device 60 switches valve 32 for a predetermined time to connect the flow path 49 and the branch flow path 50, and also switches valve 33 to connect the branch flow path 50 to tube 14, thereby sampling the reagents in flow path 49 into tube 14.

[0047] The control device 60 can control the amount of reagent sampled by controlling the time it takes to switch the flow path with the valve 32. In other words, if the liquid delivery speed of the pump 31 is known in advance, the desired amount of reagent can be accurately sampled into the tube 14 by adjusting the time it takes to connect the flow path 49 and the branched flow path 50. <Obtaining the weight of metabolites and microorganisms> Figure 4 shows the skims used to obtain the weights of metabolites and microorganisms from the culture medium of the cell culture device 100. Figure 4 shows skim 1000 for obtaining the weights of metabolites and skim 2000 for obtaining the weights of microorganisms. Skim 1000 and skim 2000 will be described below.

[0048] (Skim 1000: Obtaining the weight of metabolites) In skim 1000, step SA1 registers the ID assigned to tube 14A. In one implementation example, this registration is performed in the information processing device 70. More specifically, when the user inputs the ID assigned to tube 14A into the input device 74 of the information processing device 70, information associating that ID with the tube to be processed (tube 14A) is registered in the memory 72.

[0049] In step SA2, a mixture of culture medium and quencher is generated in tube 14A. More specifically, in the sampling device 1, the culture medium from the cell culture device 100 and the quencher (metabolic reaction cessation solution) from the reagent tank 34 are supplied to tube 14A. Step SA2 is an example of a quenching process to stop the metabolic reaction of microorganisms in the culture medium. After that, tube 14A is transported to the pretreatment device 2.

[0050] In step SA3, centrifugal force is applied to the culture medium (mixture with quencher) in tube 14A by the centrifugation mechanism 4 of the pretreatment device 2.

[0051] In step SA4, the liquid removal mechanism 5 detects the solid-liquid interface in tube 14A and removes the supernatant.

[0052] In step SA5, the reagent supply mechanism 6 supplies reagents for extracting microbial metabolites into tube 14. Subsequently, the stirring mechanism 7 stirs the mixture in tube 14A.

[0053] In step SA6, the extraction mechanism 8 heats and shakes the mixture in tube 14A, and then a portion of the mixture is extracted.

[0054] In step SA7, the extracted mixture is centrifuged to obtain the supernatant, which is then centrifuged again in step SA8. After centrifugation in step SA8, the supernatant in tube 14A is transferred to tube 14X and sent to the liquid chromatograph-mass spectrometer 3. In the information processing device 70, the ID associated with tube 14A is transferred to tube 14X.

[0055] In step SA9, mass spectrometry is performed on the solution in tube 14X using the liquid chromatograph-mass spectrometer 3, thereby quantitatively analyzing the metabolites in the culture medium introduced into tube 14A.

[0056] The information processing device 70 uses the quantitative analysis results of metabolites in the culture medium introduced into tube 14A to determine the weight of metabolites in the culture medium within tube 14A. In one implementation example, the weight of metabolites in the culture medium within tube 14A is determined based on the ratio of the peak area corresponding to the metabolite to the peak area corresponding to an internal reference substance (e.g., 2-isopropylmalic acid) in the mass chromatogram obtained for the culture medium in tube 14A (the solution in tube 14X). The information processing device 70 registers the weight of the metabolite in memory 72, associating it with the ID of tube 14A.

[0057] (Skim2000: Obtaining the weight of microorganisms) In Skim 2000, step SB1 registers the ID assigned to tube 14B. In one implementation example, this registration is performed in the information processing device 70. More specifically, when the user inputs the ID assigned to tube 14B into the input device 74, the information processing device 70 registers information in memory 72 that associates that ID with the tube to be processed (tube 14B).

[0058] In memory 72, the ID of tube 14B is associated with the ID of tube 14A. More specifically, tube 14B is supplied with culture medium from the same cell culture device 100 that supplied the culture medium to tube 14A. In the analysis system 700, the IDs of tube 14A and tube 14B, which are supplied with culture medium from the same cell culture device 100, are associated with each other.

[0059] In step SB2, the weight of tube 14B is measured and registered in memory 72. Subsequently, culture medium is supplied to tube 14B in the sampling device 1. The culture medium supplied to tube 14B is from the same cell culture device 100 as the culture medium supplied to tube 14A in step SA1 of skim 1000. In other words, culture medium is supplied to both tube 14A and tube 14B from the same cell culture device 100. After that, tube 14B is transported to the pretreatment device 2.

[0060] In step SB3, centrifugal force is applied to the culture medium in tube 14B by the centrifugation mechanism 4 of the pretreatment device 2.

[0061] In step SB4, the liquid removal mechanism 5 detects the solid-liquid interface in tube 14B, and the supernatant is transferred to another tube (tube 14Y). A different ID is registered for tube 14Y than for tube 14B, and the supernatant transferred to tube 14Y is subjected to culture medium component analysis (step SB6).

[0062] In step SB5, the weight of the pellets remaining in tube 14B after the transfer of the supernatant in step SB4 is measured. In one implementation, the user measures the weight of tube 14B containing the pellets and inputs the measurement result to the information processing device 70 via the input device 74. The information processing device 70 obtains the weight of the pellets by subtracting the weight of tube 14B registered in step SB2 from the input weight. The information processing device 70 then registers the weight of the pellets as the weight of the microorganisms in memory 72, associated with the ID of tube 14B. In another implementation, the information processing device 70 has a mechanism (not shown) for measuring the weight of tube 14B containing the pellets, measures the weight of tube 14B using this mechanism, obtains the weight of the pellets by subtracting the weight of tube 14B registered in step SB2 from the measured weight, and then registers the weight of the pellets as the weight of the microorganisms in memory 72, associated with the ID of tube 14B.

[0063] <Indicators related to the weight of metabolites> In the analysis system 700, the information processing device 70 obtains the weight of metabolites for each tube and outputs an index for the weight of the metabolites. The index for the weight of metabolites is an index that represents the amount of metabolites produced per unit weight of microorganisms.

[0064] To derive this index, the information processing device 70 supplies the same amount (volume) of culture medium from a cell culture apparatus 100 into two tubes, identifies the weight of metabolites in the culture medium supplied to one tube, identifies the weight of microorganisms in the culture medium supplied to the other tube, and calculates the product of the weight of metabolites and the reciprocal of the weight of microorganisms. The information processing device 70 obtains the calculated product value as the above index.

[0065] Figure 5 is a flowchart of the process for outputting an index of metabolite weight. In one implementation example, the process shown in Figure 5 is performed by the CPU 71 executing a given program in the information processing device 70. The process flow is described below.

[0066] In step S10, the information processing device 70 reads the weight of a metabolite associated with a certain tube (tube 14A) from the memory 72. In one implementation example, the weight of the metabolite may be obtained according to the skim 1000 in Figure 4 and registered in the memory 72.

[0067] In step S20, the information processing device 70 reads the weight of the microorganism associated with tube 14B from the memory 72. In step S20, tube 14B is associated with tube A in step S10 in the memory 72. That is, the weight of the metabolite read in step S10 and the weight of the microorganism read in step S20 relate to the same culture medium provided from the cell culture device 100.

[0068] In step S30, the information processing device 70 calculates an index as the product of the weight of the metabolite read in step S10 and the reciprocal of the weight of the microorganism read in step S20.

[0069] In step S40, the information processing device 70 displays the index calculated in step S30 on the display 75. After that, the information processing device 70 terminates the process shown in Figure 5.

[0070] In the process shown in Figure 5 described above, the index for metabolite weight is derived as the product of the weight of the metabolite identified in the culture medium in the container (cell culture device 100) and the reciprocal of the weight of the microorganism identified in the said culture medium. This index corresponds to the value derived by normalizing the metabolite weight by the weight of the microorganism. Furthermore, this index represents the amount of metabolite produced per unit weight of microorganism. Therefore, this index can be said to be equivalent to information for handling the weight of a metabolite while taking into account the environment in which the metabolite was produced (the amount or number of microorganisms in the culture medium).

[0071] In the process described with reference to Figure 5, the index is calculated and displayed. Note that "display" is just one example of how the index can be output. The information processing device 70 may also output the calculated index by transmitting it to an external device.

[0072] <Amount of liquid supplied for gravimetric analysis of microorganisms and amount of liquid supplied for gravimetric analysis of metabolites> Figure 6 shows the volume of liquid supplied to the tubes for gravimetric analysis of microorganisms and the volume of liquid supplied to the tubes for gravimetric analysis of metabolites in the analysis system 700. The vertical axis in Figure 6 represents the volume of liquid supplied to the tubes.

[0073] The graph in Figure 6 shows the results for each of the 10 groups (A1 to E2). For each group, the liquid was supplied to the tube six times. The graph in Figure 6 shows the maximum and minimum volumes of liquid supplied to the tube for each group. The results in Figure 6 are for the case where pure water was used as an example of the liquid.

[0074] Each group has a different target volume for the liquid supplied to the tube. The target volume for groups A1 and A2 is 2 mL, for groups B1 and B2 it is 1 mL, for groups C1 and C2 it is 0.5 mL, for groups D1 and D2 it is 0.2 mL, and for groups E1 and E2 it is 0.1 mL.

[0075] The numbers assigned to each group indicate different purposes for supplying liquid from the cell culture device 100 to the tubes in the analysis system 700. "1" indicates that liquid was supplied from the cell culture device 100 to the tube (tube 14B) for the purpose of weighing microorganisms (skim 2000 in Figure 4). "2" indicates that liquid was supplied from the cell culture device 100 to the tube (tube 14B) for the purpose of weighing metabolites (skim 1000 in Figure 4).

[0076] In the graph in Figure 6, group A1 includes a maximum value of 2.00g and a minimum value of 1.95g. Group A2 also includes a maximum value of 2.00g and a minimum value of 1.93g. In other words, the maximum and minimum values ​​are almost the same for group A1 and group A2. From this, it can be said that the volume of liquid supplied to tube 14B for gravimetric measurement of microorganisms for group A1 is approximately the same as the volume of liquid supplied to tube 14A for gravimetric measurement of metabolites for group A2.

[0077] Furthermore, in the graph in Figure 6, the maximum and minimum values ​​are almost identical between group B1 and group B2. In addition, the maximum and minimum values ​​are almost identical between group C1 and group C2, group D1 and group D2, and group E1 and group E2.

[0078] Therefore, in the analysis system 700, if the target volume is the same, there is a correlation between the volume of liquid supplied to tube 14B for measuring the weight of microorganisms and the volume of liquid supplied to tube 14A for measuring the weight of metabolites, which are approximately the same. Thus, in the analysis system 700, the weight of metabolites is effectively normalized by the weight (or reciprocal of the weight) of microorganisms measured using the same culture medium supplied from the same cell culture device 100 used to measure the weight of the metabolites.

[0079] In one implementation, in the analysis system 700, in order to make the amount of liquid supplied to the tubes uniform, the control device 60 may stop stirring by the stirring bar 111 (or reduce the rotational speed for stirring) for a given period immediately before supplying the culture medium from the cell culture apparatus 100 to the tubes 14 (tubes 14A, 14B). In another implementation, the information processing device 70 may instruct the control device 60 to stop such stirring (or reduce the rotational speed for stirring), and the control device 60 may stop stirring (or reduce the rotational speed) in accordance with the instruction. If the given period is too long, it may cause uneven culture in the culture medium supplied to the tubes, and if it is too short, it may hinder the uniformity of the amount due to the inclusion of bubbles in the culture medium supplied to the culture medium to supply oxygen. In this sense, the given period is, in one example, about 2 to 10 minutes, preferably 3 to 7 minutes, and more preferably 3 minutes.

[0080] [Aspect] Those skilled in the art will understand that the above-described exemplary embodiments are specific examples of the following embodiments.

[0081] (Clause 1) An information provision method according to one embodiment is a method for providing information on metabolites, comprising the steps of: obtaining the weight of a metabolite identified in the culture medium in a container; obtaining the weight of a microorganism identified in the culture medium in the container; deriving an index relating to the weight of the metabolite using the weight of the metabolite and the reciprocal of the weight of the microorganism; and outputting the index.

[0082] According to the information provision method described in paragraph 1, information is provided for handling the weight of metabolites in a manner that takes into account the environment in which the metabolites were produced.

[0083] (Paragraph 2) In the information provision method described in Paragraph 1, the step of deriving the indicator may include deriving the indicator by calculating the product of the weight of the metabolite and the reciprocal of the weight of the microorganism.

[0084] According to the information provision method described in paragraph 2, the above indicators can be easily calculated. (3) In the information provision method described in paragraph 1 or 2, the weight of the metabolite may be specified for the culture medium after it has been subjected to a quenching treatment to stop the metabolic reaction of the microorganism.

[0085] According to the information provision method described in paragraph 3, it is possible to avoid the production of metabolites proceeding after the culture medium has been separated from the container for the purpose of determining the weight of the metabolites.

[0086] (Clause 4) An analytical system according to one embodiment is an analytical system that provides information on the weight of microbial metabolites, and comprises an information processing device, the information processing device being configured to obtain the weight of identified metabolites in a microbial culture medium in a container, obtain the weight of identified microorganisms in the culture medium, derive an index relating to the weight of the metabolites using the weight of the metabolites and the reciprocal of the weight of the microorganisms, and output the index.

[0087] According to the analysis system described in paragraph 4, information is provided for handling the weight of metabolites in a manner that takes into account the environment in which the metabolites were produced.

[0088] (Clause 5) In the analytical system described in paragraph 4, the index may be derived by calculating the product of the weight of the metabolite and the reciprocal of the weight of the microorganism.

[0089] According to the analysis system described in Section 5, the above indicators can be easily calculated. (Clause 6) In the analytical system described in paragraph 4 or 5, the weight of the metabolite may be specified with respect to the culture medium after quenching treatment to stop the metabolic reaction of the microorganism.

[0090] According to the analytical system described in Section 6, the production of metabolites can be avoided after the culture medium has been separated from the container for the purpose of determining the weight of the metabolites.

[0091] (Clause 7) A program according to one embodiment is a program executed by a computer that provides information on metabolites, the computer may be instructed to perform the steps of: obtaining the weight of a metabolite identified in the culture medium in a container; obtaining the weight of a microorganism identified in the culture medium in the container; deriving an index relating to the weight of the metabolite using the weight of the metabolite and the reciprocal of the weight of the microorganism; and outputting the index.

[0092] According to the program described in paragraph 7, information is provided for handling the weight of metabolites in a manner that takes into account the environment in which the metabolites were produced.

[0093] (Clause 8) In the program described in paragraph 7, the step of deriving the index may include deriving the index by calculating the product of the weight of the metabolite and the reciprocal of the weight of the microorganism.

[0094] According to the program described in Section 8, the above indicators can be easily calculated. (Clause 9) In the program described in paragraph 7 or 8, the weight of the metabolite may be specified with respect to the culture medium after a quenching treatment has been performed to stop the metabolic reaction of the microorganism.

[0095] According to the program described in paragraph 9, the production of metabolites can be avoided after the culture medium has been separated from the container for the purpose of determining the weight of the metabolites.

[0096] The embodiments disclosed herein should be considered in all respects to be illustrative and not restrictive. The scope of this disclosure is indicated by the claims rather than by the description of the embodiments above, and all modifications within the meaning and scope of the claims are intended to be included. [Explanation of Symbols]

[0097] 1 Sampling device, 2 Pretreatment device, 3 Liquid chromatograph mass spectrometer, 4 Centrifugal separation mechanism, 5 Liquid removal mechanism, 6 Reagent supply mechanism, 7 Stirring mechanism, 8 Extraction mechanism, 10 Pretreatment system, 12 Holding unit, 13 Motor, 14 Tube, 20 Culture medium sampling mechanism, 21, 31 Pump, 22, 23, 32, 33 Valve, 25 Filter, 26 Washing solution tank, 27 Waste liquid tank, 30 Reagent sampling mechanism, 34 Reagent tank, 41, 42, 49, 50 Flow channels, 43, 45, 47 Outlet channels, 44, 46, 48 Inlet channels, 60 Control device, 70 Information processing device, 100 Cell culture device, 700 Analysis system.

Claims

1. A method for providing information on metabolites, The steps include supplying the culture medium in the container to the first tube, The steps include obtaining the weight of metabolites identified by a liquid chromatograph-mass spectrometer in the culture medium supplied to the first tube, The steps include supplying the culture medium in the container to a second tube different from the first tube, The method comprises the step of obtaining the weight of microorganisms identified by the liquid chromatograph-mass spectrometer in the culture medium supplied to the second tube, The weight of the metabolite is determined for the culture medium after it has been quenched to stop the metabolic reaction of the microorganism. The weight of the microorganism is determined for the culture medium that has not undergone the quenching treatment. The method of providing the aforementioned information further includes, The steps include: deriving an index relating to the weight of the metabolite using the reciprocal of the weight of the metabolite and the weight of the microorganism; The step of outputting the aforementioned indicator, The step of deriving the aforementioned index includes deriving the index by calculating the product of the weight of the metabolite and the reciprocal of the weight of the microorganism.

2. The information provision method according to claim 1, wherein the second tube is supplied with the same amount of culture medium as the amount supplied to the first tube.

3. The information-providing method according to claim 1 or 2, wherein the quenching treatment includes mixing a drug that stops the metabolic reaction of microorganisms in the culture medium with the culture medium.

4. An analytical system that provides information on the weight of microbial metabolites, Information processing device and A sampling device, Equipped with a liquid chromatograph-mass spectrometer, The sampling device is The culture solution of microorganisms in the container is supplied to a first tube and a second tube different from the first tube. The aforementioned information processing device is The culture medium supplied to the first tube, after being subjected to a quenching treatment to stop the metabolic reaction of the microorganisms, is used to obtain the weights of metabolites identified by the liquid chromatograph-mass spectrometer. The culture medium supplied to the second tube, which has not undergone the quenching treatment, is used to obtain the weight of microorganisms identified by the liquid chromatograph-mass spectrometer. Using the weight of the metabolite and the reciprocal of the weight of the microorganism, an index relating to the weight of the metabolite is derived. It is configured to output the aforementioned indicator, The aforementioned index is derived by calculating the product of the weight of the metabolite and the reciprocal of the weight of the microorganism, in an analytical system.

5. The analytical system according to claim 4, wherein the second tube is supplied with the same amount of culture medium as the amount supplied to the first tube.

6. The analytical system according to claim 4 or 5, wherein the quenching treatment includes mixing a drug that stops the metabolic reactions of microorganisms in the culture medium with the culture medium.

7. A program executed by a computer that provides information about metabolites, To the aforementioned computer, The steps include supplying the culture medium in the container to the first tube, The steps include obtaining the weight of metabolites identified by a liquid chromatograph-mass spectrometer in the culture medium supplied to the first tube, The steps include supplying the culture medium in the container to a second tube different from the first tube, The process involves performing the step of obtaining the weight of microorganisms identified by the liquid chromatograph-mass spectrometer in the culture medium supplied to the second tube, The weight of the metabolite is determined for the culture medium after it has been quenched to stop the metabolic reaction of the microorganism. The weight of the microorganism is determined for the culture medium that has not undergone the quenching treatment. The aforementioned computer further, The steps include: deriving an index relating to the weight of the metabolite using the reciprocal of the weight of the metabolite and the weight of the microorganism; The step of outputting the aforementioned indicator is performed, A program that derives the index by calculating the product of the weight of the metabolite and the reciprocal of the weight of the microorganism.

8. The program according to claim 7, wherein the second tube is supplied with the same amount of culture medium as the amount supplied to the first tube.

9. The program according to claim 7 or 8, wherein the quenching process includes mixing a drug into the culture medium to stop the metabolic reactions of microorganisms in the culture medium.

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