Fluorometric titrator

• US 4,476,095 A
• Filed: 04/12/1974
• Issued: 10/09/1984
• Est. Priority Date: 04/12/1974
• Status: Expired due to Term

• Current Assignee: Charles A. Bell, Charles S. Jacobs Jr, Robert L. Scott, Samuel Jr. Nejame
• Original Assignee: Charles A. Bell, Charles S. Jacobs Jr, Robert L. Scott, Samuel Jr. Nejame
• Inventors: Scott, Robert L., Bell, Charles A., Nejame, Samuel Jr., Jacobs, Charles S. Jr.
• Primary Examiner(s): Marcus, Michael S.
• Application Number:
  US05/460,297
• Time in Patent Office: 3,833 Days
• Field of Search: 436/172, 436/163, 436/51, 422/75
• US Class Current: 422/75
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 2021/6443 Fluorimetric titration
• CPC Class Codes: G01N 21/79 Photometric titration
• CPC Class Codes: G01N 35/1095 for supplying the samples to flow-through analysers for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00
• CPC Class Codes: Y10T 436/116664 with automated titrator

Abstract

A fluorometric titrator has means to transfer a pipette between first and second positions and a buret for a chelating agent has a reversible drive powered either by a pulse generator or an oscillator. Valve controlled means are operable to place the buret in communication with the pipette or a reservoir. In the first position of the pipette, a precise sample volume is aspirated into it by the buret, then driven by the oscillator and in its second position and with the buret drive reversed but still powered by the oscillator, the sample volume is discharged from the pipette into a cuvette in the fluorometer. On the resulting fluorescence the fluorometer output drives the pulse generator to dispense chelating agent and to operate a counter until a selected end point is reached. The pipette is then returned to its second position with the buret drive again operated by the oscillator and during the return, the buret is refilled. Both before the sample volume is aspirated, and before that volume is discharged, backlash is removed from the buret drive. The effect of hysteresis on the photocell of the fluorometer is minimized by light control also enabling the response time of the titrator to be accelerated.

Claims

1. A fluorometric titrator comprising a device operable to aspirate and subsequently discharge a sample, reversible means to transfer said device between first and second positions, sample control means to aspirate a precise sample volume into said device when in said first position and to discharge that volume in said second position, a fluorometer including a cuvette to receive the discharged volume and to hold a reagent that fluoresces when said sample volume is discharged therein, means to deliver chelating agent into said cuvette, a pulse generator controlled by the output of said fluorometer in response to resulting fluorescence to operate the delivery means for the chelating agent at a decreasing rate as the fluorescence is reduced and to produce an indication at the value of a chemical contained in the sample.

2. The fluorometric titrator of claim 1 in which the end point detector is also operable to control the return of the device to its first position.

3. The fluorometric titrator of claim 1 in which the fluorometer has a chamber opening both upwardly and downwardly, a holder detachably attached to the chamber closes the bottom of the opening and has a recess, and the cuvette is shaped and dimensioned to fit the recess thereby to be held in a predetermined position within the chamber.

4. The fluorometric titrator of claim 1 and means operable to render the end point of the detector substantially constant, at least after the first of a series of titrations.

5. The fluorometric titrator of claim 4 and means operable, at least after the first of a series of titrations to provide for each subsequent titration starting point having a substantially constant relation to the previous end point.

6. The fluorometric titrator of claim 5 in which the output circuit of the fluorometer includes the end point detector and means to increase the intensity of the lamp as the fluorescence is quenched to maintain the luminosity on the sensor of the fluorometer substantially constant.

7. The fluorometric titrator of claim 6 in which the means provide that the detector circuit always has a predetermined initial voltage at the start of a titration.

8. The fluorometric titrator of claim 1 in which the sample control means includes a pump and a reversible drive to operate said pump in either an aspirating or a delivering direction and means operable before the aspiration of a precise sample volume and before the discharge thereof to remove backlash from said pumping means.

9. The fluorometric titrator of claim 8 in which the pump and drive for the sample control means is also the delivery means for the chelating agent.

10. The fluorometric titrator of claim 9 in which the pump drive includes a reversible stepper motor and a transmission, and the means powered by the output of the fluorometer includes a pulse generator to drive the stepper motor during a titration and the sample control means includes an oscillator as an alternative drive for the stepper motor and means to time the operation of the oscillator in both the aspiration and the discharge of the precise sample volume.

11. The fluorometric titrator of claim 9 and means isolating the device from the pumping means during the operation of the backlash-removing means.

12. The fluorometric titrator of claim 11 and means operable during the return of said device and while it is connected to the pumping means to aspirate, into the device, a predetermined volume of air to provide a bubble as a barrier between the chelating agent and a sample and operable during the discharging stage to discharge said;

bubble after the discharge of the sample volume.

13. The fluorometric titrator of claim 1 in which the delivery means is a pump operable both to aspirate and discharge fluids and to contain a sufficient volume of the chelating agent for at least one titration and has a drive including a reversible stepper motor, and there is a reservoir for the chelating agent, conduit means effecting communication between the delivery means, the device, and the reservoir and including a valve and reversible means to shift the valve between first and second positions, in the first, the device in communication with the pump and in the second, the reservoir and the pump in communication with each other, an oscillator as an alternative drive for said motor, and first and second timers as alternative means to power said oscillator, the sample control means includes the first timer, and means to energize said second timer to operate said pumping means to remove backlash from the delivery means before the sample concontrol means is operated in either position and also to aspirate chelating agent into the delivery means in the second valve position.

14. The fluorometric titrator of claim 13 and a chamber having top, front, side and end walls joined in a leakproof manner, the bottom of said chamber having at least one port, the pump, valve and device are located in the chamber, and the reversible drives therefor each supported by an appropriate one of said walls on the outside of said chamber.

15. The fluorometric titrator of claim 13 and a control circuit including switching means providing, after the operation of the sample control means to aspirate a precise sample volume and with the stepper motor set to operate the delivery means in a dispensing direction and the valve in its first position, three stages of operation, in the first, the transfer means operated to transfer the device to its second position and with the valve in its first position at the second position of the device and the sample control means then operated, in the second, the stepper motor driven by the pulse generator until the end point is detected, the direction of the motor then reversed to operate the delivery means in an aspirating direction and the valve is set in its second position, and in the third stage, energizing the stepper motor to aspirate the chelating agent from the reservoir, and then return the valve to its first position at least when the device is in its first position.

16. The fluorometric titrator of claim 15 in which the transfer means and the valve shifting means each include first and second switches for its first and second positions, respectively, and the delivery means also includes a switch operable when the pump contains an insufficient volume of the chelating agent, means operable during said first stage to set the valve in its second position with its second switch then operative to effect the energization of the second timer thereby to pump back to the reservoir enough chelating agent to remove backlash from the delivery means, the first switch of the pumping means then operable to effect the reverse operation of the valve shifting means before the second switch of the transfer means is closed, at the end of said second stage, the end point detector effects the direction of the stepper motor to drive the delivery means in an aspirating direction and the shifting of the valve into its second position thereby to energize the second timer unless and until the delivery means switch is operated, and the first switch of the transfer means, when closed, stopping the operation of the titrator.

17. The fluorometric titrator of claim 16 in which the delivery means also includes a second switch operable when the pumping means contains a predetermined maximum volume of the chelating agent, said second switch, when operated effects the setting of the valve in its first position, the first switch of the valve shifting means effecting the energization of the second timer whereby air is aspirated into the device as a barrier between the chelating agent and the next sample volume to be aspirated, the first switch of the transfer means when closed holding the second timer inoperative while the device is in its first position, and the second timer includes means operable when the device is in its second position to operate the second timer at the end of the interval provided by the first timer.

18. The fluorometric titrator of claim 16 in which the first position of the valve includes means, operable when the stepper motor is driven to effect the operation of the delivery means in an aspirating direction, to halt the transfer means between its two positions while the second timer is energized.

19. The fluorometric titrator of claim 16 and additional switch controlled means operable, when the switch thereof is closed, to set the direction the stepper motor will operate to drive the delivery means in an aspirating direction when energized and set the valve in its first position.

20. The fluorometric titrator of claim 1 in which value producing means is a readout responsive to the operation of the fluorometer to provide a value for the titration of a predetermined volume of a known value of a sample and means operable should the readout value be different than the known value to vary the operation of the sample control means so that it aspirates to vary the aspirated sample volume to provide a readout value to match said known value.

21. The fluorometric titrator of claim 20 and second means operable to vary the size of the aspirated sample volume independently of the means enabling readout and known values to be matched.

22. A fluorometric titrator comprising a device operable to aspirate and subsequently discharge a sample transferrable between first and second positions, sample control means operable to aspirate a precise sample volume into said device when in said first position and to discharge that volume with the device in said second position, a fluorometer including a cuvette to receive the discharged volume and to hold a reagent that fluoresces when the sample volume is discharged therein, means to deliver chelating agent into said cuvette, a pulse generator controlled by the output of said fluorometer in response to resulting fluorescence to operate the delivery means for the chelating agent at a decreasing rate as the fluorescence is reduced and to produce an indication of the value of a chemical contained in the sample.

23. The fluorometric titrator of claim 22 in which the sample control means includes pumping means in communication with the device, a reversible drive in control of said pumping means thereby to provide both for the aspiration of the sample volume and its discharge, an oscillator the output pulses of which said drive, and adjustable means to time the operation of the drive by the oscillator first with the drive operable to activate the pumping means in an aspirating direction and then in a discharging direction.

24. The fluorometric titrator of claim 23 and means to convert the output pulse output of the oscillator as determined by the timing means to at least one other and predetermined different number of output pulses whereby a different precise sample volume is aspirated and delivered without changing the timing means.

25. A fluorometric titrator comprising a device operable to aspirate and subsequently discharge a sample to contain a selected sample volume, reversible means to transfer said device between first and second positions, the first position the sample receiving position, sample control means to discharge that volume when the device is in said second position, a fluorometer including a cuvette to receive the discharged volume and to hold a reagent that fluoresces when said sample volume is discharged therein, means to deliver chelating agent into said cuvette, a pulse generator controlled by the output of said fluorometer in response to resulting fluorescence to operate the delivery means for the chelating agent at a decreasing rate as the fluorescence is reduced and to produce an indication of the value of a chemical contained in the sample.

26. The fluorometric titrator of claim 25 in which the reversible transfer means includes an arm and a reversible drive including a shaft in support of one end of the arm and operable to swing the arm through an arc from one position to the other and the device is supported by the other end of the arm to extend downwardly as the arm swings between the two positions of the device, and the transfer means includes means providing a predetermined angular disposition of said device at each of said positions.

27. The fluorometric titrator of claim 26 in which the device positioning means includes a gear train mounted on said arm, the device connected to the last gear of said train to turn therewith, a fixed gear coaxial with the shaft, and the first gear of the train in mesh with the fixed gear.

28. A fluorometric titrator comprising a fluorometer which yields an output, a cuvette for a fluorometric indicator solution, pumping means for a chelating agent including a delivery conduit in communication with the cuvette and operable to deliver a chelating agent thereto after a sample has been introduced into the cuvette to cause fluorescence, and delivery means controlled by the output of the fluorometer in response to the resulting fluorescence to operate the pumping means at a decreasing rate as the fluorescence is reduced, said pump means including a cylinder having an outlet at one end, a slidable piston having an axial bore opening towards the other end of the cylinder and including a fixed nut located at the open end of said bore, and the delivery means includes a motor, a drive shaft rotated thereby and including a threaded portion threaded through said nut and extending into said bore, the nut and shaft threads being of the micrometer type.

29. The fluorometric titrator of claim 28 in which the pumping means has a cylinder for the chelating agent and the piston includes an end portion that is a free fit in the cylinder and is of a uniform diameter throughout its cylinder-entering length thereby to function on a positive displacement basis.

30. A fluorometric titrator comprising a fluorometer which includes a sensor provided with an output circuit, a cuvette, a stirrer in the cuvette, the cuvette for a fluorescent indicator solution, pumping means for a chelating agent including a conduit in communication with the cuvette and operable to deliver the chelating agent thereto after a sample has been introduced into the cuvette that causes fluorescence, a drive for said pumping means including a stepper motor, the output circuit of the sensor of the fluorometer including a pulse generator operable to drive said motor and operating at a pulse-producing rate that decreases as the fluorescence is quenched and a detector opening said circuit when the fluorescence is reduced to a value selected as the end point of the test, said circuit also including means operable to adjust said end point to compensate for end point drift at least after the first of a series of titrations.

31. The fluorometric titrator of claim 30 in which the circuit includes voltage regulating means operable to increase the intensity of the fluorometer lamp as the fluorescence is quenched to maintain the luminosity on the sensor substantially constant.

32. The fluorometric titrator of claim 30 in which the end point detector is a voltage comparator, the output circuit of the fluorometer is connected thereto, a lead including a capacitor and an amplifier is connected to the comparator to provide the standard voltage with which the fluorometer output is compared with the comparator ending the titration when the voltages match, and switch means operable at the end of a titration to connect the output circuit to the capacitor to charge it whereby the next titration will have the voltage standard of the previous end point including any change or drift in the previous end point.

33. A fluorometric titrator comprising a fluorometer which yields an output and including a cell, a lamp and a cuvette between said cell and said lamp to receive a selected sample volume and adapted to hold a reagent that fluoresces when said sample volume is received therein, means to deliver chelating agent into said cuvette, a pulse generator controlled by the output of said fluorometer in response to resulting fluorescence to operate the delivery means for the chelating agent at a decreasing rate as the fluorescence is reduced, and means in control of said lamp and operable to maintain the cell luminosity at a constant level at least between the titrations.

34. The titrator of claim 33 in which the light control means is also operable in response to the decreasing cell luminosity to increase the lamp voltage to maintain, during a titration, the luminosity constant as the fluorescence decreases.

35. The titrator of claim 33 and means operable by the pulse generator to produce an indication of the value of a chemical contained in the sample.

36. In combination, a fluorometer having a chamber that opens downwardly and includes a holder detachably attached to the chamber to close the chamber opening and formed with a flat-surfaced recess, and a cuvette molded from a transparent material and dimensioned to fit the chamber and within the recess and be held in a predetermined position therein by the holder.