Automatic mixing and dilution methods and apparatus for online characterization of equilibrium and non-equilibrium properties of solutions containing polymers and/or colloids

• US 20040004717A1
• Filed: 05/21/2003
• Published: 01/08/2004
• Est. Priority Date: 11/13/1996
• Status: Abandoned Application

• Current Assignee: Administrators Of The Tulane Educational Fund (ADMINISTRATORS|DAVID H. COY)
• Original Assignee: Administrators Of The Tulane Educational Fund (ADMINISTRATORS|DAVID H. COY)
• Inventors: Reed, Wayne F.
• Application Number:
  US10/442,676
• Publication Number:
  US 20040004717A1
• Time in Patent Office: Days
• US Class Current: 356/338
• CPC Class Codes: B01D 15/34 Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
• CPC Class Codes: G01N 15/0211 Investigating a scatter or diffraction pattern
• CPC Class Codes: G01N 15/075 by optical means
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 2011/0046 In situ measurement during mixing process
• CPC Class Codes: G01N 2015/0092 Monitoring flocculation or agglomeration
• CPC Class Codes: G01N 2015/0216 from fluctuations of diffraction pattern
• CPC Class Codes: G01N 2015/025 Methods for single or grouped particles
• CPC Class Codes: G01N 2015/0693 by optical means, e.g. by integrated nephelometry
• CPC Class Codes: G01N 2021/4711 Multiangle measurement
• CPC Class Codes: G01N 2021/4716 Using a ring of sensors, or a combination of diaphragm and sensors; Annular sensor
• CPC Class Codes: G01N 2021/4719 using a optical fibre array
• CPC Class Codes: G01N 2021/513 Cuvettes for scattering measurements
• CPC Class Codes: G01N 21/49 within a body or fluid
• CPC Class Codes: G01N 21/51 inside a container, e.g. in an ampoule G01N21/53 takes precedence
• CPC Class Codes: G01N 21/53 within a flowing fluid, e.g. smoke
• CPC Class Codes: G01N 2201/0218 Submersible, submarine
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/74 Optical detectors measurement of intensity, velocity, spectral content, polarisation, or phase of infrared, visible or ultraviolet light G01J
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: G01N 35/085 : Flow Injection Analysis
• CPC Class Codes: G01N 35/1097 : characterised by the valves valves in general F16K

Abstract

A method involving the automatic, online dilution of polymer and/or colloid solutions, such that, when the diluted polymer stream flows through suitable detectors, non-equilibrium processes, such as polymerization, degradation and aggregation, can be monitored. The dilution involves a reacting or stock solution of polymer and/or colloid, and at least one solvent. The online dilution technique can also be used to assess the effects of solvent quality and other solutes on polymer/colloid characteristics and reactions, and also permits equilibrium characterization of polymers/colloids by making a single stock solution of the polymer/colloid.

A device is developed that is capable of automatically and continuously extracting fluid from a polymer-containing vessel and mixing this with a solvent such that the final fluid is dilute enough that single particle light scattering, spectrophotometric and other measurements can be made on it. Whereas many sampling and dilution devices exist, the novelty of this invention consists in its ability to deal with very high viscosities, including those laden with bubbles, and to introduce only a short delay time between sampling and measurement. The device is ideally suited for situations where the viscosity of the polymer-containing vessel changes over a wide range during the course of a reaction; e.g. polymerization, polymer degradation, aggregation, and others. Furthermore, provision is made for modular conditioning stages, such as changing solvent conditions, evaporating monomer, filtering, etc. The amount of sample actually withdrawn for measurement is very low, normally on the order of 0.25 ml to 5 ml per hour. The device can also vary the dilution factor either automatically or manually during operation.

Claims

1. Apparatus for static light scattering for absolute macromolecular characterization, comprising:

a submersible probe for use with at least one photodetector and a computer electronically connected to the photodetector, the computer being programmed for analyzing data from static light scattering for performing absolute macromolecular characterization, the probe being submersible in a fluid to be sampled, the probe comprising;

a light source;

light detection means secured in a fixed position relative to the light source;

transmission means for transmitting light from the light detection means to the photodetector, the transmission means being of a sufficient length and flexibility to allow the submersible probe to be submersed in the fluid to be sampled without submersing the photodetector.

2. The apparatus of claim 1, further comprising the photodetector and the computer.

3. The apparatus of claim 1, comprising a plurality of photodetectors.

4. The apparatus of claim 1, wherein the probe comprises:

a ring member having a channel into which sample fluid enters upon immersion;

means for securing the light source in the ring member; and

means for securing the light detection means in the ring member.

5. The apparatus of claim 4, wherein the ring member includes a beam dump.

6. The apparatus of claim 4, wherein the ring member is made of an opaque material.

7. The apparatus of claim 1, wherein the light detection means comprises fiber optic light conduits.

8. The apparatus of claim 1, further comprising means for removably attaching the light transmission means to the photodetector.

9. The apparatus of claim 1, further comprising a harness for securing the light transmission means to minimize damage to the light transmission means.

10. The apparatus of claim 2, wherein the size range of detectability is about 20 Angstroms to about 100 microns.

11. The apparatus of claim 2, wherein the detectable range of particles is from about 500 g/mole to about 10¹⁴ g/mole.

31. A method of performing absolute macromolecular characterization with static light scattering using the apparatus of claim 1, the method comprising:

submersing the probe of claim 1 in a sample fluid;

optically connecting the probe of claim 1 with a photodetector, the photodetector being electronically connected to a computer, the computer being programmed for analyzing data from static light scattering for performing absolute macromolecular characterization, the probe being submersible in a fluid to be sampled;

using the computer, analyzing data from static light scattering for performing absolute macromolecular characterization of the sample fluid in which the probe is submersed.

12. Apparatus for static light scattering, comprising:

a submersible probe for use with at least one photodetector and a computer electronically connected to the photodetector, the computer being programmed for analyzing data from static light scattering for performing characterization of a fluid, the probe being submersible in a fluid to be sampled, the probe comprising;

a ring member;

a light source secured in the ring member;

light detection means secured in the ring member in a fixed position relative to the light source;

transmission means for transmitting light from the light detection means to the photodetector, the transmission means being of a sufficient length and flexibility to allow the submersible probe to be submersed in the fluid to be sampled without submersing the photodetector.

13. The apparatus of claim 12, further comprising the photodetector and the computer.

14. The apparatus of claim 12, comprising a plurality of photodetectors.

15. The apparatus of claim 12, wherein the probe comprises:

a ring member having a channel into which sample fluid enters;

means for securing the light source in the ring member; and

means for securing the light detection means in the ring member.

16. The apparatus of claim 15, wherein the ring member includes a beam dump.

17. The apparatus of claim 15, wherein the ring member is made of an opaque material.

18. The apparatus of claim 12, wherein the light detection means comprises fiber optic light conduits.

19. The apparatus of claim 12, further comprising means for removably attaching the light transmission means to the photodetector.

20. The apparatus of claim 12, further comprising a harness for securing the light transmission means to minimize damage to the light transmission means.

21. The apparatus of claim 13, wherein the size range of detectability is about 20 Angstroms to about 100 microns.

22. The apparatus of claim 13, wherein the detectable range of particles is from about 500 g/mole to about 10¹⁴ g/mole.

23. Apparatus for static light scattering, comprising:

a plurality of interchangeable probes for use with at least one light detector and a computer electronically connected to the photodetector, the computer being programmed for analyzing data from static light scattering for performing characterization of a fluid, each probe comprising;

a ring member;

a light source secured in the ring member;

light detection means secured in the ring member in a fixed position relative to the light source;

transmission means for transmitting light from the light detection means; and

means for removably connecting the transmission means to the photodetector.

24. The apparatus of claim 23, wherein the transmission means allows the probe to move relative to the photodetector when the transmission means is connected to the photodetector.

25. The apparatus of claim 23, further comprising the photodetector and the computer.

26. The apparatus of claim 23, wherein at least one of the probes is submersible.

27. The apparatus of claim 23, wherein at least one of the probes has connectors to allow fluid conduits to be attached thereto.

28. The apparatus of claim 23, wherein at least one of the probes has a receptacle for holding sample fluid.

29. The apparatus of claim 23, wherein at least one of the probes has means for receiving a receptacle for holding sample fluid.

30. The apparatus of claim 23, wherein at least one of the probes has a handle.

58. The apparatus of claim 23, further comprising a plurality of photodetectors.

32. A method of performing absolute macromolecular characterization with static light scattering, the method comprising:

optically contacting a probe with a sample fluid containing a substance being studied, the probe having a scattering volume containing a small enough number of large scattering particles to not prevent absolute macromolecular characterization of the substance being studied;

optically connecting the probe to a photodetector, the photodetector being electronically connected to a computer with an interface, the computer being programmed for analyzing data from static light scattering for performing absolute macromolecular characterization;

using the computer, analyzing data from static light scattering for performing absolute macromolecular characterization of the sample fluid in optical contact with the probe, while electronically separating out scattering bursts from large scattering particles in the sample fluid, allowing hence the large scattering particles to also be counted and characterized, wherein;

the photodetector and the interface operate at a rate fast enough to electronically resolve the bursts.

33. The method of claim 32, wherein the rate is at least 2 Hz.

34. The method of claim 32, wherein the number of large scattering particles is less than 100 per scattering volume.

35. The method of claim 32, further comprising the step of counting and characterizing the large scattering particles.

36. A method of conducting absolute macromolecular characterization in real time in a polymerization reaction, comprising:

(a) diluting a sample fluid to be sampled to a concentration of such that interparticle effects do not dominate the scattering behavior;

(b) irradiating the diluted sample fluid with incident light;

(c) measuring light scattered from the diluted sample fluid with at least one photodetector and a computer electronically connected to the photodetector, the computer being programmed for analyzing data from static light scattering for performing absolute macromolecular characterization.

37. The method of claim 36, wherein light is transmitted from the diluted sample fluid to the photodetector with a submersible probe comprising:

a light source for providing the incident light;

light detection means secured in a fixed position relative to the light source; and

transmission means for transmitting light from the light detection means to the photodetector, the transmission means being of a sufficient length and flexibility to allow the submersible probe to be submersed in the fluid to be sampled without submersing the photodetector.

38. The method of claim 36, wherein the dimensionless quantity 2A₂cM_w is not greater than 10.

39. The method of claim 36, wherein the dimensionless quantity 2A₂cM_w is not greater than 2.

40. The method of claim 36, wherein the photodetector has a scattering volume containing a small enough number of large scattering particles to not prevent absolute macromolecular characterization of the substance being studied;

using the computer, analyzing data from static light scattering for performing absolute macromolecular characterization of the sample fluid in optical contact with the probe, while electronically separating out scattering bursts from large scattering particles in the sample fluid, allowing hence the large scattering particles to also be counted and characterized, wherein;

the photodetector and the interface operate at a rate fast enough to electronically resolve the bursts.

59. The method of claim 36, wherein the diluting of the sample fluid occurs on-line.

60. The method of claim 59, wherein the dimensionless quantity 2A2cMw is not greater than 2.

41. Apparatus for static light scattering for absolute macromolecular characterization, comprising:

a probe for use with at least one photodetector and a computer electronically connected to the photodetector, the computer being programmed for analyzing data from static light scattering for performing absolute macromolecular characterization, the probe comprising;

a light source;

light detection means secured in a fixed position relative to the light source;

transmission means for transmitting light from the light detection means to the photodetector, the transmission means being of a sufficient length and flexibility to allow the probe to be used remote from the photodetector.

42. The apparatus of claim 41, wherein transmission means is 6 inches-100 feet long.

43. Apparatus for static light scattering, comprising:

an interchangeable probe for use with at least one light detector and a computer electronically connected to the photodetector, the computer being programmed for analyzing data from static light scattering for performing characterization of a fluid, the probe comprising;

a ring member;

a light source secured in the ring member;

light detection means secured in the ring member in a fixed position relative to the light source;

transmission means for transmitting light from the light detection means; and

means for removably connecting the transmission means to the photodetector.

44. The apparatus of claim 43, wherein the transmission means allows the probe to move relative to the photodetector when the transmission means is connected to the photodetector.

45. The apparatus of claim 43, further comprising the photodetector and the computer.

46. The apparatus of claim 43, wherein the probe is submersible.

47. The apparatus of claim 43, wherein the probe has connectors to allow fluid conduits to be attached thereto.

48. The apparatus of claim 43, wherein the probe has a receptacle for holding sample fluid.

49. The apparatus of claim 43, wherein the probe has means for receiving a receptacle for holding sample fluid.

50. The apparatus of claim 43, wherein the probe has a handle.

51. The apparatus of claim 43, wherein the light detection means are placed at scattering angles of from about 10°

to about 170°

.

52. The apparatus of claim 43, wherein the ring member includes a beam dump.

53. The apparatus of claim 43, wherein the ring member is made of an opaque material.

54. The apparatus of claim 43, wherein the transmission means is of a sufficient length and flexibility to allow the probe to be used remote from the photodetector.

55. The apparatus of claim 43, further comprising means for using the apparatus for absolute macromolecular characterization.

56. The apparatus of claim 55, further comprising means for counting particles simultaneously.

57. The apparatus of claim 43, wherein the ring member contains a sampling cavity that can be directly contacted with sample fluid, which cavity can hold sample fluid which is introduced via flow, pipeting, immersion or other means, and which fluid can either remain stationary in the ring or flow through it.

61. A method of making a real-time measurement of a reaction, interaction or process occurring in a solution containing polymers and/or colloids, comprising:

(a) automatically diluting and/or mixing online at least two separate solutions, at least a first solution containing polymers and/or colloids and at least a second solution containing a solvent or a solution containing other types of polymers and/or colloids, to create a diluted and/or mixed solution;

(b) measuring characteristics of the diluted and/or mixed solution;

(c) determining from the measurements made in step (b) characteristics of the reaction, interaction or process occurring in first solution containing polymers and/or colloids.

62. The method of claim 61, wherein at least three solutions are mixed together.

63. The method of claim 61, wherein a light scattering detector is used to determine the relative molecular mass of a polymer during a polymerization reaction.

64. The method of claim 61, wherein a suitable concentration detector is used to simultaneously measure the concentration of solutes in the mixed solution.

65. The method of claim 61, further comprising using a light scattering detector to determine, online, the absolute weight averaged molecular mass Mw of a polymer as it is produced in a polymerization reaction.

66. The method of claim 65, wherein a flow type viscometer is placed inline, so that reduced viscosity can be determined simultaneously with Mw, and a measure of polydispersity can hence be formed, online, by combining the values of reduced viscosity and Mw.

67. The method of claim 61, wherein a light scattering detector is used to monitor the relative degradation of a polymer solution caused by enzymes or other chemical agents, radiation, or heat.

68. The method of claim 67, wherein the absolute Mw of the degrading polymer solution is monitored.

69. The method of claim 68, wherein a concentration detector is also used to monitor the absolute Mw of the degrading polymer solution.

70. The method of claim 63, wherein the first and second solutions can interact at certain concentrations, and the interaction is detected online by the light scattering detector.

71. The method of claim 70, wherein the Mw of the interacting species is measured online.

72. The method of claim 71, wherein a concentration detector is also used to measure the Mw of the interacting species online.

73. The method of claim 70, wherein a viscometric detector is used to measure online changes in relative viscosity due to interactions.

74. The method of claim 71, wherein a viscometric detector is used to determine online the reduced viscosity.

75. The method of claim 72, wherein a viscometric detector is used to determine online the reduced viscosity.

76. The method of claim 63, wherein the first and second solutions can interact at certain concentrations, and the interaction is detected online by a turbidity sensing device.

77. The method of claim 61 wherein HTDSLS is used to recognize and characterize large scattering particles in the diluted and/or mixed solution, whether such particles be impurities or integral components of the polymer/colloid solution being characterized.

78. The method of claim 62, wherein:

the polymer/colloid concentration in the diluted and/or mixed solution is held constant,

the second solution contains a solvent and is produced by mixing of two or more additional reservoirs and is made to vary, and

the effects of the second solution on the polymers/colloids is measured online.

79. The method of claim 61, wherein:

the online dilution is used to dilute a polymer/colloid solution, and

absolute macromolecular characterization is performed using light scattering and concentration detectors.

80. The method of claim 79, further comprising using a viscometric detector to detect viscosity.

81. Apparatus including a device capable of automatically and continuously diluting and/or mixing a high viscosity fluid in at least two stages, wherein a first mixing of the high viscosity fluid and a diluent occurs to create a first mixed stream, and at least a second mixing occurs in which the first mixed solution is mixed with a diluent or diluents in one or more stages to create a mixed stream for measurement.

82. The apparatus of claim 81, wherein the fluid ranges in viscosity from 50 to 5,000,000 cP.

83. The apparatus of claim 81, wherein the viscosity of the fluid increases from less than 1 cP to over 5,000,000 cP during the course of a reaction, or decreases from over 5,000,000 cP to less than 1 cP.

84. The apparatus of claim 83, wherein the viscosity change occurs over an interval of no less than a minute and not more than 48 hours.

85. The apparatus of any one of claims 81-84, wherein the relative viscosity of the fluid increases anywhere from a factor of around 300 to a factor of around 10,000,000.

86. The apparatus of any one of claims 81-85, wherein the dilution factor is in the range of around 2 to 50,000.

87. The apparatus of any one of claims 81-86, wherein the diluted or mixed material is continuously withdrawn for analysis at a rate ranging from around 0.001 to 1000 ml/minute.

88. The apparatus of any one of claims 81-87, wherein the diluted or mixed material flows through a train of detectors in order to analyze the contents of the vessel containing the viscous liquid.

89. The apparatus of any one of claims 81-88, wherein the high viscosity fluid is contained in a polymerization reactor.

90. The apparatus of any one of claims 81-89, wherein the vessel containing the high viscosity fluid is a fermentation reactor.

91. The apparatus of any one of claims 81-90, wherein the high viscosity fluid is a biological or bioactive polymer, such as a protein, polysaccharide, pharmaceutical agent, etc.

92. The apparatus of any one of claims 81-91, further comprising a light scattering detector for analysis of the mixed stream for measurement.

93. The apparatus of any one of claims 81-92, further comprising a concentration detector for analysis of the mixed stream for measurement.

94. The apparatus of claim 93, wherein the concentration detector is from the group consisting of an ultraviolet/visible spectrometer, a refractometer, and an evaporative light scattering device.

95. The apparatus of any one of claims 81-94, further comprising a viscometer for analysis of the mixed stream for measurement.

96. The apparatus of any one of claims 81-95, further comprising a near infra-red detector for analysis of the mixed stream for measurement.

97. The apparatus of any one of claims 81-96, further comprising a solution conductivity detector for analysis of the mixed stream for measurement.

98. The apparatus of any one of claims 81-97, further comprising a primary pump and a liquid containing vessel containing high viscosity fluid, and wherein the primary pump of the device recirculates viscous liquid to the liquid containing vessel, and a fraction of this recirculating flow is diverted, either continuously, or at intervals, for mixing or diluting.

99. The apparatus of any one of claims 81-98, wherein the mixing or diluting of the viscous liquid takes place in a low pressure mixing chamber.

100. The apparatus of any one of claims 81-99, further comprising detectors and debubbling means, wherein any bubbles are exhaled in a mixing/diluting stage and are absent in the mixed stream for measurement reaching the detectors.

101. The apparatus of any one of claims 81-100, including an inlet and an outlet, and further comprising at least one sample conditioning stage between the inlet and outlet.

102. The apparatus of claim 101, wherein there are a plurality of sample conditioning stages between the inlet and outlet.

103. The apparatus of any one of claims 81-102, wherein the mixed stream for measurement is dilute enough to enable useful measurements to be taken by at least one measuring device from the group consisting of a light scattering detector, a concentration detector, an ultraviolet/visible spectrometer, a refractometer, an evaporative light scattering device, a viscometer, and a conductivity detector.

104. The apparatus of any one of claims 81-102, wherein the dilution factor can be held constant, or can be varied either manually or automatically during use.

105. The apparatus of any one of claims 81-104, wherein a microprocessor-containing device (e.g. a microcomputer) is used to control one or more pumps, such that the dilution factor and/or detector feed flow rate can be automatically controlled.

106. A method of automatically and continuously diluting and/or mixing high viscosity fluids comprising using the apparatus of any one of claims 81-105.

107. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases from a factor of 1,000 to a factor of 100,000.

108. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases a factor of at least 100.

109. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases a factor of at least 500.

110. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases a factor of at least 1,000.

111. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases a factor of at least 5,000.

112. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases a factor of at least 50,000.

113. The invention of any one of claims 81-106, wherein the relative viscosity of the fluid increases or decreases a factor of at least 500,000.

114. The invention of any one of claims 81-113, wherein at least two mixing stages occur after the first mixed stream is created.

115. The invention of any one of claims 81-113, wherein at least three mixing stages occur after the first mixed stream is created.

116. The invention of any one of claims 81-113, wherein at least four mixing stages occur after the first mixed stream is created.

117. The invention of any one of claims 81-113, wherein at least five mixing stages occur after the first mixed stream is created.

118. The inventions substantially as shown and described herein.