Automatic analysis method and apparatus for enzyme reaction

Claim 2

1 . An automatic analysis method of enzyme reaction for automatically carrying out all the steps of weighing operation of a sample, adding operation of solvent to the sample, dissolving operation of the sample in the solvent, adding operation of the solvent into the sample solution, injecting operation of the diluted sample solution into a reaction tube, adding operation of enzymes into the reaction tube to produce a reaction mixture, the moving operation of the reaction tube onto preservation vessel, filtering operation of the reaction mixture into the reaction tube to produce a filtrate, concentrating operation of the filtrate, injecting operation of the filtrate into the analysis apparatus, and processing operation of the analysis data with respect to the filtrate.

Claim 3

An automatic analysis method of enzyme reaction, in accordance with Claim 1, wherein the weighing of a sample, adding of a solvent, dissolution are automatically carried out by all the steps of grasping a sample tube retained in a rack with a chuck provided on a movable arm of an automatic robot for moving the sample tube to an electronic balance, a solvent dispenser, a dissolution vessel, adding the solvent predetermined in accordance with the weighing value of sample after weighing operation of the substance by the electronic balance, placing the sample tube into a dissolution vessel after the addition of a solvent to dissolve the sample, moving the sample tube with the dissolved sample in the rack for dilution provided in a reaction apparatus of the next step, the addition of solvent, injection into the reaction tube, addition of the enzyme reaction mixture are carried out by the steps of grasping each nozzle of a diluted solution dispenser, a sampling pipetter and an enzyme reaction mixture dispenser with the chuck of the automatic robot and moving the nozzle and injecting into the sample tube retained in the rack for dilution use and the reaction tube retained in the rack placed in the incubator for performing injecting operation, adding dilution solvent from a diluted solution dispenser into the sample tube in a dilution rack to perform the diluting operation into given times, injecting diluted solution of the sample tube from the dispenser to inject it into the reaction tube, sequentially injecting the enzymes and factors necessary for enzyme reaction into the reaction tube at the intervals of given time to start the reaction, grasping the reaction tube by the robot to move onto the preservation vessel after the reaction, grasping the reaction tube retained in the preservation vessel by the robot to place the reaction tube in the position of a reaction mixture sampling needle disposed in a filtration/concentration/injection (FCI) apparatus, filtering reaction mixture through a filtration unit after sampling operation of reaction mixture with a sampling needle, feeding the eluate into a column for concentration use after the adsorption of a filtrate onto a column for the concentration use to inject into an analytical column of a high performance liquid chromatograph so that the obtained data is processed with a processing apparatus, and the operation of the robot is automatically controlled in accordance with a program input into the computer to automatically perform all the steps from weighing to analysis.

Claim 4

An automatic analysis method of enzyme reaction in accordance with Claim 1, wherein the enzyme is lipoxygenase.

Claim 5

4 . An automatic analysis method of enzyme reaction in accordance with Claim 1, wherein the enzyme is cyclooxygenase.

Claim 6

5 . An automatic analysis method of enzyme reaction in accordance with Claim 1, wherein solution for enzyme reaction of the cell including the lipoxygenase or the cyclo - oxygenase, the arachidonic acid, and the ethanol is provided by adding sequentially, at the intervals of the given time.

Claim 7

An automatic weighing and dissolving method for samples, comprising the steps of retaining in a rack a plurality of sample tubes for free taking-out operations in different positions on the periphery of an automatic robot, an electronic balance, a solvent adding apparatus, a dissolution vessel, so that the sample tube in the rack into which a sample is thrown is freely grasped with a chuck disposed on a movable arm of the automatic robot to sequentially move it automatically onto the electronic balance, the solvent adding apparatus and the dissolution vessel, and employing a computer for controlling the operation of said robot, setting a solvent adding amount calculated in accordance with the weight of the sample weighed by an electronic balance, adding the solvent into the sample tube, for automatically controlling each of the apparatuses such as dissolution vessels into which the sample tube after the addition of solvent is placed.

Claim 8

An automatic weighing and dissolving method for samples, comprising the steps of retaining in a rack a plurality of sample tubes for free taking-out operations in different positions on the periphery of an automatic robot, an electronic balance, a solvent adding apparatus, a dissolution vessel, so that the sample tube in the rack into which a sample is thrown is freely grasped with a chuck disposed on a movable arm of the automatic robot to sequentially move it automatically onto the electronic balance, the solvent adding apparatus and the dissolution vessel, and employing a computer for controlling the operation of said robot, setting a solvent adding amount calculated in accordance with the weight of the sample weighed by an electronic balance, adding the solvent into the sample tube, for automatically controlling each of the apparatuses such as dissolution vessels into which the sample tube after the addition of solvent is placed.

Claim 9

7 . An automatic dilution reacting method comprising the steps of retaining in a rack with a plurality of sample tubes, a dilution dispenser, a sampling pipetter, a incu- b ator having a plurality of reaction tubes disposed in the incubator, a nozzle washing vessel disposed on positions different on the periphery of an automatic robot, nozzles mounted respectively through the flexible tubes on each of the dispensers, these nozzles being detachably placed on the nozzle stand provided within the operation range of the robot while each of the nozzles is freely grasped with a chuck provided on a movable arm of the robot so that the nozzle may move into the nozzle washing vessel, the opening of the sample tube in a rack or into the opening of the reaction tube in an incubator, and automatically controlling, by a controller, the robot in a manner that the diluted sample solution is pipetted into the sample tube in a rack from a dilution dispenser and a diluted sample is pipetted from the sample tube by the sampling pipetter to be pipetted into the reaction tube in the incubator, and the reaction mixture is pipetted from a reaction mixture sampling pipetter into a dilution sample in the incubator.

Claim 10

7 . An automatic dilution reacting method comprising the steps of retaining in a rack with a plurality of sample tubes, a dilution dispenser, a sampling pipetter, a incu- b ator having a plurality of reaction tubes disposed in the incubator, a nozzle washing vessel disposed on positions different on the periphery of an automatic robot, nozzles mounted respectively through the flexible tubes on each of the dispensers, these nozzles being detachably placed on the nozzle stand provided within the operation range of the robot while each of the nozzles is freely grasped with a chuck provided on a movable arm of the robot so that the nozzle may move into the nozzle washing vessel, the opening of the sample tube in a rack or into the opening of the reaction tube in an incubator, and automatically controlling, by a controller, the robot in a manner that the diluted sample solution is pipetted into the sample tube in a rack from a dilution dispenser and a diluted sample is pipetted from the sample tube by the sampling pipetter to be pipetted into the reaction tube in the incubator, and the reaction mixture is pipetted from a reaction mixture sampling pipetter into a dilution sample in the incubator.

Claim 11

8 . An automatic filtration/concentretion/injecting ( FCI ) method comprising the steps of feeding by a syringe pump, sampling a mixture into a sampling solution pipetter from a sample tube with a sampling needle, filtering the pipetted sample mixture by a filtering unit of a filter-paper continuous winding type, controlling a valve with a sampling loop thereon to feed the filtered sample mixture and a valve with a concentration column thereon, feeding by a water pump the sample solution in the loop of the sampling loop valve, together with water, into a concentration column of the valve with a concentration column on it, and feeding through a valve feeding the eluate into the concentration column to inject into an analytical column the sample solution located within the concentration column.

Claim 12

8 . An automatic filtration/concentretion/injecting ( FCI ) method comprising the steps of feeding by a syringe pump, sampling a mixture into a sampling solution pipetter from a sample tube with a sampling needle, filtering the pipetted sample mixture by a filtering unit of a filter-paper continuous winding type, controlling a valve with a sampling loop thereon to feed the filtered sample mixture and a valve with a concentration column thereon, feeding by a water pump the sample solution in the loop of the sampling loop valve, together with water, into a concentration column of the valve with a concentration column on it, and feeding through a valve feeding the eluate into the concentration column to inject into an analytical column the sample solution located within the concentration column.

Claim 13

An automatic analysis apparatus of enzyme reaction, comprising a means for weighing a sample, a means for adding solvent to the sample, a means for dissolving the sample in the solvent, a means for adding solvent to the dissolved sample solution, a means for injecting diluted sample solution into a reaction tube, a means for adding enzymes into the reaction tube, a means for moving the reaction tube onto a preservation vessel, a means for filtering the reaction mixture in the reaction tube, a means for concentrating the filtrate, a means for injecting into an analysis apparatus, and a means processing the analysis data.

Claim 14

An automatic analysis apparatus of enzyme reaction, comprising a means for weighing a sample, a means for adding solvent to the sample, a means for dissolving the sample in the solvent, a means for adding solvent to the dissolved sample solution, a means for injecting diluted sample solution into a reaction tube, a means for adding enzymes into the reaction tube, a means for moving the reaction tube onto a preservation vessel, a means for filtering the reaction mixture in the reaction tube, a means for concentrating the filtrate, a means for injecting into an analysis apparatus, and a means processing the analysis data.

Claim 15

An automatic analysis method of enzyme reaction, in accordance with Claim 9, wherein the means for performing weighing operation, adding solvent and performing dissolution comprises a rack for retaining many sample tubes on the different periphery of a robot to be automatically controlled, an electronic balance, a solvent dispenser, a dissolution vessel with an ultrasonicator apparatus thereon so that the sample tube in the rack is grasped with chuck disposed on the movable arm of a robot to move it onto the electronic balance, the solvent adding means, the dissolution vessel, the dilution reaction means for a resolved sample comprises a rack for dilution use for retaining many sample tubes in the different position on the periphery of the robot to be automatically controlled, a incubator provided with the rack for retaining many reaction tubes, a dilution dispenser, a sampling pipetter, a plurality of dispensers for injecting enzymes, a preservation vessel provided with a rack for retaining the reaction tube, so that a nozzle of each injector is grasped with a chuck of the robot to move it onto the sample tube and the reaction tube to perform the injection, the reaction tube in the reaction vessel is grasped with the chuck of the robot to move it onto the rack of the preservation vessel, the filtration/concentration/injection (FCI) apparatus comprises a sampling needle for sampling the reaction mixture from the reaction tube through the operation by a syringe pump, a filtration unit of a filter paper continuous winding type for filtering the sampled reaction mixture, a concentration column for concentrating the filtrate, a valve for feeding the eluate into the concentration column to inject it into an analytical column, each of these apparatuses being so disposed that the filtration, concentration, injection is performed on one line.

Claim 16

An automatic weighing and dissolution apparatus for samples comprising a rack for retaining a plurality of sample tubes for free taking-out operations in the different positions on the periphery of an automatic robot, an electronic balance, a solvent adding apparatus, a dissolution vessel, so that the sample tube in the rack into which the sample is thrown is freely grasped with a chuck disposed on a movable arm of the automatic robot to sequentially move it automatically onto the electronic balance, the solvent adding apparatus and the dissolution vessel, and a computer for controlling the operation of said robot, setting the solvent adding amount calculated in accordance with the weight of the sample weighed by the electronic balance, adding the solvent into the sample tube, for automatically controlling each of the apparatuses such as dissolution vessels into which the sample tube after the addition of solvent is placed.

Claim 17

An automatic weighing and dissolution apparatus for samples comprising a rack for retaining a plurality of sample tubes for free taking-out operations in the different positions on the periphery of an automatic robot, an electronic balance, a solvent adding apparatus, a dissolution vessel, so that the sample tube in the rack into which the sample is thrown is freely grasped with a chuck disposed on a movable arm of the automatic robot to sequentially move it automatically onto the electronic balance, the solvent adding apparatus and the dissolution vessel, and a computer for controlling the operation of said robot, setting the solvent adding amount calculated in accordance with the weight of the sample weighed by the electronic balance, adding the solvent into the sample tube, for automatically controlling each of the apparatuses such as dissolution vessels into which the sample tube after the addition of solvent is placed.

Claim 18

An automatic weighing and dissolving apparatus for samples in accordance with Claim 11, wherein a horizontal multi-joint type robot is used as the automatic robot.

Claim 19

An automatic weighing and dissolving apparatus for samples in accordance with Claim 11, wherein a plural kinds of solvent is set for the solvent adding apparatus.

Claim 20

An automatic weighing and dissolving apparatus for samples in accordance with Claim 11, wherein a heater is mounted in the dissolution vessel with an ultrasonicator apparatus on it so that the heater is adapted to be controlled by the computer.

Claim 21

An. automatic weighing and dissolving apparatus for samples in accordance with Claim 11, wherein the taking-in or taking-out of the sample tube with respect to the electronic balance is adapted to be controlled by the computer.

Claim 22

An automatic weighing and dissolving apparatus for samples in accordance with Claim 11, wherein the amount of solvent and diluted solution into the sample is added in response to the weight of the sample weighed by the electronic balance.

Claim 23

An automatic dilution reacting apparatus comprising a rack for retaining a plurality of sample tubes, a dilution dispenser, a sampling pipetter, an incubator having a plurality of reaction tubes disposed in the vessel, a nozzle washing vessel disposed on the positions different on the periphery of an automatic robot, nozzles mounted respectively through flexible pipes on each of the dispensers, these nozzles being detachably placed on a nozzle stand provided in the operation range of the robot while each of the nozzles is freely grasped with a chuck provided on a movable arm of the robot so that it may move into the opening of the sample tube in the nozzle washing vessel and a rack or into the opening of the reaction tube in the incubator, and a controller for automatically controlling the robot so provided that the diluted sample is pipetted from the sample tube by the sampling pipetter to be injected into the reaction tube in the incubator, and the reaction mixture is injected from the reaction mixture dispenser into the diluted sample in the incubator.

Claim 24

An automatic dilution reacting apparatus comprising a rack for retaining a plurality of sample tubes, a dilution dispenser, a sampling pipetter, an incubator having a plurality of reaction tubes disposed in the vessel, a nozzle washing vessel disposed on the positions different on the periphery of an automatic robot, nozzles mounted respectively through flexible pipes on each of the dispensers, these nozzles being detachably placed on a nozzle stand provided in the operation range of the robot while each of the nozzles is freely grasped with a chuck provided on a movable arm of the robot so that it may move into the opening of the sample tube in the nozzle washing vessel and a rack or into the opening of the reaction tube in the incubator, and a controller for automatically controlling the robot so provided that the diluted sample is pipetted from the sample tube by the sampling pipetter to be injected into the reaction tube in the incubator, and the reaction mixture is injected from the reaction mixture dispenser into the diluted sample in the incubator.

Claim 25

An automatic dilution reacting apparatus in accordance with Claim 17, wherein a preservation vessel is provided on the periphery of the robot, and the reaction tube is adapted to be grasped with the chuck of the robot after the reaction by the incubator to be moved into the preservation vessel.

Claim 26

19 . An automatic weighing and dissolving apparatus for samples in accordance with Claim 17, wherein a horizontal multi-joint type robot is used as the automatic robot.

Claim 27

20 . An automatic dilution reacting apparatus in accordance with Claim 17, wherein a digital diluter pipetter is used as each of the dispensers, and a coil-shaped teflon tube is used as a coupling flexible pipe so that the nozzle is integrally formed at the tip end of the tube.

Claim 28

21 . An automatic dilution reacting apparatus in accordance with Claim 17, wherein a dissolution vessel with a magnetic stirrer on it is used as the incubator.

Claim 29

22 . An automatic dilution reacting apparatus in accordance with Claim 17, wherein a plurality of dispensers for the reaction mixture different in kind are provided as the reaction mixture sampling pipetter, and the reaction mixture different in kind are sequentially injected into one reaction tube at the intervals of the given time to cause plural kinds of reactions.

Claim 30

An automatic dilution reacting apparatus in accordance with Claim 22, wherein at least enzyme solution is used as the reaction mixture.

Claim 31

An automatic filtration/concentration/injection ( FCI ) apparatus comprising a syringe pump for feeding sampling solution pipetted from a sample tube with a sampling needle, a filter unit for filtering by a filter paper continuous winding type the pipetted sample solution, a valve with a sampling loop on it to which the filtered sample solution is fed, a valve with a concentration column on it, a water pump for feeding the sample solution in the loop of the valve with a sampling loop, together with water, into a concentration column of a valve with the concentration column thereon, and a valve for feeding eluate into the concentration column to inject into an analytical column for sample solution in the concentration column.

Claim 32

An automatic filtration/concentration/injection ( FCI ) apparatus comprising a syringe pump for feeding sampling solution pipetted from a sample tube with a sampling needle, a filter unit for filtering by a filter paper continuous winding type the pipetted sample solution, a valve with a sampling loop on it to which the filtered sample solution is fed, a valve with a concentration column on it, a water pump for feeding the sample solution in the loop of the valve with a sampling loop, together with water, into a concentration column of a valve with the concentration column thereon, and a valve for feeding eluate into the concentration column to inject into an analytical column for sample solution in the concentration column.