WO2007069005A1

WO2007069005A1 - Automatic sampling and bioluminometric analysis device

Info

Publication number: WO2007069005A1
Application number: PCT/IB2006/001644
Authority: WO
Inventors: Giorgio Bellini
Original assignee: Star Ecotronics S.R.L.
Priority date: 2005-12-16
Filing date: 2006-06-07
Publication date: 2007-06-21
Also published as: ITMI20052407A1; EP1960755A1

Abstract

A device (100) for automatizing fluid sampling and bio- luminometric analysis comprises a console (1) carrying its own tools, a luminometer (2) , a horizontally sliding handling set to which a vertically sliding head (200) is associated, said vertically sliding head (200) lodging on it a metering syringe, sterilizing and cleaning means for said metering syringe (19) , storing, withdrawing and location means suitable to receive the samples to be analyzed, a multifunction cylinder (4) provided with a recirculation path and a withdrawing well for the fluid samples, and a number reagent withdrawing wells, and electronic means for processing data from said luminometer.

Description

Title: "Automatic sampling and bioluminometric analy¬

sis device"

The present invention has as an object a device for automatizing fluid sampling and bioluminometric

analysis.

As known, bioluminescence is the most used method to determine sterility or to assess the bacterial charge of a fluid. Such method relies on measuring the amount of light emitted by the reaction occurring when a reagent, namely the luciferin-luciferase, comes into contact with the ATP (adenosine triphosphate) released by the bacteria contained in the fluid being analysed.

The light emission is measured by a photonic count system based on a photo-multiplier and a digital count line.

In prior art systems for the bioluminometric analysis of fluids, the fluid sample is manually withdrawn by a person in charge and carried to the

analysis lab or, at best, it is analysed in situ by means of a portable bioluminometer .

Subsequently, in dependence of the result ob-

tained by the analysis, the bacterial charge values of the analysed fluid are adjusted in a second step.

However, it should be noted that the operations performed according to the above-said procedure do not allow an optimal running of the entire work cycle related to such analysis.

Therefore, an object of the present invention is

to solve the above-said problems by producing a device for automatizing sampling and bioluminometric analysis of fluids, which device allows to automati- cally control the bacterial charge of a fluid.

A further object of the present invention is to

produce an automatic sampling and bioluminometric analysis device that, in dependence of the values obtained by the luminometric analysis, automatically acts on fluid by metering biocides as a corrective action.

A further object of the present invention is to

produce an automatic sampling and bioluminometric analysis device that provides alarm signals to the operators upon reaching predetermined values .

Another object of the present invention is to produce an automatic fluid sampling and bioluminomet¬

ric analysis device that automatically modifies the fluid sampling rate according to the values obtained by previously performed analysis. Finally, an object of the present invention is to produce an automatic fluid sampling and bioluminomet- ric analysis device that assesses the bacterial charge of the analyzed fluid in order to compare it with a predetermined minimum threshold value.

Such objects are reached by a device for automatizing fluid sampling and bioluminometric analysis according to claim 1, to which we refer for brevity.

The invention is hereafter described in details, in an exemplary and not limiting way, with reference to the drawings attached, wherein:

Fig. 1 shows a front view of the automatic fluid sampling and bioluminometric analysis device accord¬

ing to the present invention; and Fig. 2 shows a front view of a multifunction recirculation cylinder pertaining to the device according to the present invention.

The automatic fluid sampling and bioluminometric

analysis device of the present invention is desig- nated as a whole, in an exemplary embodiment, by the reference number 100.

The automatic fluid sampling and bioluminometric analysis device 100, as shown in fig. 1, is comprised of a console or cabinet 1 preferably made of steel, which supports the number of components forming the device 100 .

In the upper part of the console 1 there are

lodged a luminometer 2, some horizontal ball guides 3 on which a head set 200 slides, and a splined shuttle (not marked in the figure) on which are fastened a cylinder provided with a recirculation path and a the

fluid sample withdrawing well 4, a container for the frozen reagent vials 5, a variable number of reagent withdrawing wells 6, 6', and a container for sterile tubes 7, from which said tubes are taken away and placed in a rotating tube carrier 20.

The head set 200, which is horizontally slidable, also provides a vertically sliding head 51 and a metering syringe 19 on it . A metering syringe slider 53 and a tube gripper 11 are also associated with the handling tower 21. A multifunction rod is associated to the head set 200 too.

In the underside of the console 1, a container 8 for collecting used tubes is lodged, which container is connected to the console 1 by means of a discharge channel 9 ending at an inlet 10 on which used tubes are dropped by a gripper 11.

In the underside of the console 1 there are also lodged an intake pipe 12 for the fluid to be ana- lyzed, a discharge pipe 13 for the fluid to be ana- lyzed, a priming pump 14, a pipe for admitting washing fluid 15, a pipe for discharging washing fluid and air 16, a pipe for admitting pressurized air 17, and a tank 18 containing a substance suited for ster- ilization.

In the backside of the console 1 (not shown in

the figure) some electric connectors to feed the device 100 and provide process interfacing, and fitments for the various pipes are lodged. The electronic components provided for feeding the device 100 and interfacing/driving motors and

feeding the electrovalves to handle the members of device 100 and the PC set are inside the console 1.

Inside the console 1 are also lodged a washing pulse pump, a number of electrovalves belonging to the hydraulic and pneumatic circuits, and a system

suited to move a horizontal handling tower 21 of the head set 200 horizontally.

The multifunction recirculation cylinder 4 per- taining to the device according to the present invention is shown in fig. 2.

In it is provided an inlet 30 in which a conduit 41 is provided to admit the sterilizing liquid from the tank 18.

Said inlet leads to a clear cylinder 36 in which are provided a conduit 40 for the liquid to be analyzed, an upper valve 33 with its own motor 31 and a pre-filter valve 34 with its own motor 32.

A filter 35 to block suspended particles is pro-

vided between said valves 33, 34.

A recirculation valve for the sample to be ana¬

lyzed is also shown in fig. 2, which valve is shown at positions Hi 38 and Lo 37.

The operation of the automatic sampling and bio- luminometric analysis device is the following.

When the device is in stand-by state, a metering syringe 19 is on the handling tower 21 of the head

set 200, which syringe, at the end of its previous operation was subjected to a complete sterilization and cleaning cycle.

In that position, syringe 19 is enclosed into a sterile room which is protected from dust and inadvertent shocks .

The priming pump 14 acting on the sewage dis- charge well 50 and connected to the intake pipe 12 causes continuous circulation of the water to be analyzed.

Two compartments are provided in the multifunction cylinder 4. In the lower part having circular

section, the water (continuously passing) moves as a whirlpool (which movement is induced by the intake pipe, which is tangential to the compartment body) thus avoiding deposition of solid agglomerates. Also

the pipes are smooth (above all the feed ones) and have a constant caliber to circumvent the same prob¬

lem.

The upper compartment of the cylinder forms the withdrawing well for the sample to be analyzed. The well communicates with the lower compartment of the

cylinder through a stainless steel mesh filter 35

(provided with 0.1 mm openings) and a conical valve that hermetically divides the two compartments.

When not in use, the withdrawing well has both the conical valve and the upper communication port of the syringe closed.

The handling tower 21 places at the multifunction cylinder 4 in order to withdraw the sample to be analyzed.

The movement of said tower is performed by an electric motor provided with an adapter and horizon¬

tal ball guides. The position references are absolute, with an acceleration and deceleration motor

range in both directions of movement.

With tower 21 placed at the multifunction cylin-

der 4, the multifunction rod extends, opening the up- per plug of the well, and the descending head 51 introduces the metering syringe 19 which, with three- five plunger travels withdraws the sample.

Then, the head 51 rises, the multifunction rod retracts and the multifunction cylinder plug closes.

The tower 21 is placed at the rotating tube car¬

rier 20.

The tube carrier 20 rotates 90° so that its inlet is directed towards a communicating ejection hole of the vial container 7.

By means of an extractor or similar means, a tube is charged on the rotating carrier 20, which is rotated so that the tube is in a vertical position.

The head 51, when going down, introduces the sy- ringe 19 into the tube (the external diameter of the syringe is smaller than the internal diameter of the tube) , and by suitably feeding the motor for a sliding movement of the syringe plunger, a slow ejection

of the sample is provided. By way of example only, one can proceed ejecting the first 100 μL of sample, such an ejection being just above the predicted total filling level of 400 μL, in order not to form air bubbles or drops scattered on the tube walls.

The sampling of the subsequent IOO μL is per- formed avoiding the multifunction cylinder sample filling first step.

Once the withdrawal is concluded, the well is

completely filled with washing/sterilization fluid through an intake pipe from the pulse pump, and the exceeding fluid, through a siphon communicating with the well at that section between the filter and the conical valve, enters a bottomless discharge pipe, allowing a perfect cleaning of the mesh filter 35 too.

Once concluded the pouring into the tube of the desired amount of sample to be analyzed, the head 51 rises and thus the tower 21 is placed back at the multifunction cylinder 4.

Then the head comes down, the cylinder is opened and a sterilization and drying step of the syringe 19 and the cylinder 4 itself is performed.

At this point, the amounts (for example 100 μL)

of one or more reagents contained into compartments 6 and required for the subsequent luminometric analysis are injected into the tube, optionally performing a sterilization and drying step of the syringe 19 after every withdrawal, as needed.

During this reaction step of the reagents into the tube, a sterilization and drying step of the sy- ringe into the cylinder 4 can be performed, always taking into consideration the reagent permanence times into the tube before the luminometric analysis, which must be constant . At the end of the sample and reagent preparation step, the tube is ready for being analyzed.

The tower 21 is placed at the location of the rotating tube carrier 20 in order to take the tube by means of the gripper 11. Then, the head 51 rises again taking the tube with it, and the tower 21 moves to the luminometer 2 and the port thereof .

The multifunction rod is withdrawn, the port of the luminometer 2 is opened and the multifunction rod enters a hole provided in the insertion carrier. By closing the port the luminometric measurement is automatically started, which allows to assess the sample bacterial concentration.

Once the analysis is concluded, the port is opened, the tube is gripped, the port of luminometer

2 is closed and the tube is then dropped through the inlet 10 of the used tube container 8.

In dependence of the analysis results, a relays outlet can, if necessary, activate biocide introduc- tion or trigger an alarm indicating the critical threshold.

Thereafter, the tower 21 of the head set 51 is placed on the location of the multifunction cylinder 4 and when the work is finished, a complete sterili- zation and cleaning cycle is performed.

It is important to note that the described device has highly adaptable characteristics which allows different uses according to the requirements of the analysis one wants to perform. Particularly, it is noted that, on the one hand the use of the device can be directed to verify that, in a production process, the bacterial charge of the final product, of one component or of one intermediate product, is below a preset limit. It is specifically provided the opportunity to meter biocides as a corrective action, to provide alarm signals to the operators, and automatically modify the analysis frequency for a more and more ac¬

curate observation. On the other hand, the device of the invention can be used to control a bacterial presence being greater than a minimum amount into an environment or

a product, which assures for example the proper operation of a purification plant or of a fermentation process. It has also been found that with the device of the invention, the following operations are automatically performed in a beneficial way:

- remote withdrawal of the sample to be analyzed by means of an immersion and continuous recirculation pump;

- sterilization/cleaning of the withdrawing well and filter thereof;

- handling of the tubes and the extractable com- partment of the luminometer;

- sample and reagent metering into the tube;

- bioluminometric analysis of the sample bacterial charge;

- timing of cycles, reagent injection and meter- ing;

- arrangement/displaying of data/statistics;

- setting of successive analysis frequency as a function of bacterial charge level;

- arrangement of external feedback for bacteri- cide introduction and critical threshold alarm;

- cleaning, sterilization and storing of the metering syringe .

From the above, it is clear that the inventive concepts described are not limited to the applicative examples shown, but they can been beneficially adapted to other similar application.

The use range of the invention is not limited to the examples shown in the above description.

The present invention can be therefore subjected

to various modifications and changes all included into the inventive concept expressed in the attached

claims, while technical details can change as needed.

Claims

1. A device for automatizing fluid sampling

and bioluminometric analysis, characterized in that it comprises a console for carrying its own tools, a luminometer, an horizontally sliding handling set to which a vertically sliding head is associated, said

vertically sliding head lodging on it a metering syringe, sterilizing and cleaning means for said metering syringe, storing, withdrawing and location means suitable to receive the samples to be analyzed, a multifunction cylinder provided with a recirculation path and a withdrawing well for the fluid samples, and a number of reagent withdrawing wells, and electronic means for processing data from said luminome- ter.

2. A device for automatizing fluid sampling and bioluminometric analysis according to claim 1, char¬

acterized in that said vertically sliding head includes at least a slider for said metering syringe, a gripper for said tubes, and that in order to charge the tubes into the luminometer, a multifunction rod is associated to said handling set.

3. A device for automatizing fluid sampling and

bioluminometric analysis according to claims 1 and 2, characterized in that a container for collecting used tubes is lodged in the underside of said console, which container is connected to said console by means of a discharge channel ending at an inlet on which used tubes are dropped by said gripper.

4. A device for automatizing fluid sampling and bioluminometric analysis according previous claims, characterized in that it provides at least an intake pipe for the fluid to be analyzed, a discharge pipe for the fluid to be analyzed, a priming pump, an pipe for admitting washing fluid, a pipe for discharging washing fluid and air, an pipe for admitting pressurized air, and a tank containing a substance suited for sterilization.

5. A device for automatizing fluid sampling and bioluminometric analysis according previous claims,

characterized in that it provides electric connectors to feed said device, means for process interfacing, and fitments for the various pipes, as well as elec¬

tronic elements to feed said device and drive the mo- tors and electrovalves to handle the device members.

6. A device for automatizing fluid sampling and bioluminometric analysis according previous claims, characterized in that said multifunction recirculation cylinder provides an inlet in which a pipe for

admitting sterilization fluid is lodged, and in which said inlet leads to a cylinder where a pipe for the liquid to be analyzed, an upper valve with its own motor and a pre-filter valve with its own motor are

lodged.

7. A device for automatizing fluid sampling and bioluminometric analysis according claim 6, charac¬

terized in that between said upper valve and said pre-filter valve it is provided a filter to block suspended particles .

8. A device for automatizing fluid sampling and

bioluminometric analysis according claim 6 or 7, characterized in that said multifunction cylinder provides a recirculation valve for the sample to be analyzed.

9. A process for fluid sampling and biolumi¬

nometric analysis, performed by the use of the device according to previous claims, characterized in that it provides a step in which the metering syringe is

subjected in advance to a sterilization and cleaning cycle and enclosed into a sterile room which is protected from dust and inadvertent shocks, a step in which the liquid to be analyzed is continuously circulated, a step in which the sample to be analyzed is withdrawn, a step in which a tube is taken and lo- cated to receive the sample to be analyzed, a step in which the well is cleaned, a step in which the syringe and cylinder are sterilized and dried, one or more steps in which the reagents required for the

next luminometric analysis are introduced into the tube, optionally performing a sterilization and drying step of said syringe after every withdrawal, and a sample luminometric analysis step after said tube has been placed into said luminometer.