US3753925A - Cerebrospinal fluid control standard - Google Patents

Cerebrospinal fluid control standard Download PDF

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Publication number
US3753925A
US3753925A US00237905A US3753925DA US3753925A US 3753925 A US3753925 A US 3753925A US 00237905 A US00237905 A US 00237905A US 3753925D A US3753925D A US 3753925DA US 3753925 A US3753925 A US 3753925A
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Prior art keywords
cerebrospinal fluid
control standard
per
glucose
determination
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US00237905A
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A Louderback
A Fontana
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Bayer Corp
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Baxter Laboratories Inc
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Assigned to TECHNICON INSTRUMENTS CORPORATION, 511 BENEDICT AVENUE, TARRYTOWN, NEW YORK 10591-6097, A CORP. OF DE. reassignment TECHNICON INSTRUMENTS CORPORATION, 511 BENEDICT AVENUE, TARRYTOWN, NEW YORK 10591-6097, A CORP. OF DE. ASSIGNMENT OF ASSIGNORS INTEREST. Assignors: COOPER LABORATORIES, INC.,
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/76—Assays involving albumins other than in routine use for blocking surfaces or for anchoring haptens during immunisation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2496/00—Reference solutions for assays of biological material
    • G01N2496/05—Reference solutions for assays of biological material containing blood cells or plasma
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/66—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood sugars, e.g. galactose
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/84—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving inorganic compounds or pH
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
    • Y10T436/104998—Glucose, ketone, nitrate standard or control
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00—Chemistry: analytical and immunological testing
    • Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
    • Y10T436/105831—Protein or peptide standard or control [e.g., hemoglobin, etc.]

Definitions

  • This invention relates to a synthetic control standard for use in the determination of cerebrospinal fluid proteins and other constituents.
  • Cerebrospinal fluid is a clear, colorless fluid which is contained in the subarachnoid spaces of the brain and spinal cord and the ventricles of the brain. It contains a few lymphocytes and has a protein content of about 15 to 45 mg. per 100 ml. (mg. percent). Under certain patient conditions, for example, brain damage, brain or spinal tumors, it is desired to assay the cerebrospinal fluid for total protein content and other parameters.
  • cerebrospinal fluids are precious specimens.
  • cerebrospinal fluid protein As with other biological fluid materials, it is desirable to employ control standards of known composition for comparison with the patients sample.
  • these control standards have comprised carefully collected and processed human cerebrospinal fluid.
  • the product is costly.
  • cerebrospinal (fluid is collected in such small quantities the material tends to deteriorate over the normal storage period required for collection of a suitable quantity for sale such as a liter or more. Enzymes in the fluid will tend to react with other constituents and product break-down products. Because of the numerous small samples which make up any collection of significant volume, the risk of bacterial contamination is greater than in ordinary blood serum collection.
  • the human cerebrospinal fluid material is diflicult to handle in processing since it does not lyophilize readily. Because its mass is relatively small, it forms a fine powder which tends to collect on the upper walls and closure of the sample bottles with a consequent loss of material upon opening the closure and reconstitution with water prior to use.
  • a synthetic control standard for the determination of cerebrospinal fluid is prepared from normal blood serum by dilution with a reagent comprising glucose and chloride ion in aqueous solution to a level whereby the total protein concentration ranges from about 30 to about 145 mg. percent. It is preferred that the diluted serum contains from about 40 to about 200 mg. percent glucose and from about to about 130 meq. per liter of chloride ion.
  • the chloride ion can be provided by use of an aqueous solution of an alkali metal chloride, preferably sodium chloride.
  • the chloride salt and the glucose preferably are analytical or reagent grade materials.
  • pre-alburnin is a special fraction obtained from blood serum and is described, for example, by Got, et al., Isolement et caractrisation dune pralbumine du serum
  • the aqueous solution is lyophilized to a dry material.
  • the lyophilized material is improved by the addition of a small amount of gum acacia prior to lyophilization, for example, on the order of about 0.1%. This improved lyophilized, dry material can be readily reconstituted with water prior to use without product loss on the container closure.
  • the synthetic cerebrospinal fluid control standard made as described herein has been found to produce much sharper bands in the electrophoretic pattern in actual use than obtained with the normal human cerebrospinal fluid. This is an added advantage of the invention in practice and enables the technician to improve the accuracy of the control procedures.
  • EXAMPLE 1 A synthetic control standard for use in the determination of cerebrospinal fluid is prepared as follows:
  • Human blood serum from which the clotting factors have been removed is diluted with an aqueous solution of glucose and sodium chloride containing 6.38 grams of NaCl and 600 mg. of glucose per liter.
  • the serum is thereby diluted to a total protein concentration of 110 mg. per ml. by admixing 65 parts by volume of the diluent solution with one part by volume of the serum.
  • To one liter of the diluted serum is then added 10 ml. of a 10% solution of gum acacia in water.
  • the resulting solution is then filled in 3 ml. aliquots into 5 ml. bottles and lyophilized.
  • the lyophilized, dry material is stored at 2 to 8 C. until required for use. In use, the dry material is reconstituted by dilution with 3 ml. of distilled water per bottle.
  • control standards prepared in Examples 1 and 2, above are used in the same manner as an unknown cerebrospinal fluid for comparison with the patients sample to provide a check on the analytical determination.
  • the method of making a synthetic control standard for use in the determination of cerebrospinal fluid constituents comprising diluting normal blood serum with an aqueous diluent containing from about to about 200 mg. per 100 ml. of glucose and from about to about 130 milliequivalents per liter of chloride ion to a level at which the total protein concentration ranges from about 30 to about 145 mg. per ml.
  • the method of claim 1 including the additional steps of admixing the liquid control standard with about 0.1% gum acacia and lyophilizing to a dry, powdered material.
  • a synthetic control standard for use in the determination of cerebrospinal fluid comprising normal blood serum diluted with an aqueous diluent containing from about 40 to about 200 mg. per 100 ml. of glucose and from about 80 to about milliequivalents per liter of chloride ion to a level at which the total protein concentration ranges from about 30 to about mg. per 100 ml.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Hematology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

A SYNTHETIC CONTROL STANDARD FOR THE DETERMINATION OF CERBROSPINAL FLUID CONSTITUENTS IS PROVIDED BY DILUTING NORMAL BLOOD SERUM WITH AN AQUEOUS SOLUTION OF GLUCOSE AND SALINE TO A TOTAL PROTEIN CONCENTRATION OF 30 TO 145 MG. PER 100 ML.

Description

United States Patent "ce 3,753,925 CEREBROSPINAL FLUID CONTROL STANDARD Allan L. Louderback, Temple City, and Anthony J.
Fontana, Glendora, Calif., assignors to Baxter Laboratories, Inc., Morton Grove, Ill. No Drawing. Filed Mar. 24, 1972, Ser. No. 237,905 Int. Cl. G01n 31 /00 US. Cl. 252-408 3 Claims ABSTRACT OF THE DISCLOSURE A synthetic control standard for the determination of cerebrospinal fluid constituents is provided by diluting normal blood serum with an aqueous solution of glucose and saline to a total protein concentration of 30 to 145 mg. per 100 ml.
This invention relates to a synthetic control standard for use in the determination of cerebrospinal fluid proteins and other constituents.
Cerebrospinal fluid is a clear, colorless fluid which is contained in the subarachnoid spaces of the brain and spinal cord and the ventricles of the brain. It contains a few lymphocytes and has a protein content of about 15 to 45 mg. per 100 ml. (mg. percent). Under certain patient conditions, for example, brain damage, brain or spinal tumors, it is desired to assay the cerebrospinal fluid for total protein content and other parameters.
The normal adult total volume of cerebrospinal fluid is about 125 and 150 ml. and it is renewed about every 3 to 4 hours. Normally, a spinal cord puncture will supply a specimen of only about 5 to ml. and repeat punctures are usually to be avoided. In an average 6 ml. specimen, 4 ml. is generally used up in laboratory analysis for the patient from which the sample is obtained, leaving only about 2 ml. per patient for retention for other purposes. In a 300 bed hospital, it generally takes on the average about 9 months to collect one liter of surplus human cerebrospinal fluid. Thus, cerebrospinal fluids are precious specimens.
In the determination of cerebrospinal fluid protein, as with other biological fluid materials, it is desirable to employ control standards of known composition for comparison with the patients sample. Heretofore, these control standards have comprised carefully collected and processed human cerebrospinal fluid. However, because of the relatively small amount of material available, the product is costly. Moreover, because cerebrospinal (fluid is collected in such small quantities, the material tends to deteriorate over the normal storage period required for collection of a suitable quantity for sale such as a liter or more. Enzymes in the fluid will tend to react with other constituents and product break-down products. Because of the numerous small samples which make up any collection of significant volume, the risk of bacterial contamination is greater than in ordinary blood serum collection. Furthermore, the human cerebrospinal fluid material is diflicult to handle in processing since it does not lyophilize readily. Because its mass is relatively small, it forms a fine powder which tends to collect on the upper walls and closure of the sample bottles with a consequent loss of material upon opening the closure and reconstitution with water prior to use.
Accordingly, it is an object of this invention to provide a synthetic control standard for use in the determination of cerebrospinal fluid.
Patented Aug. 21, 1973 It is another object of this invention to provide a synthetic cerebrospinal fluid control standard which has improved lyophilization handling characteristics over the naturally occurring material. Other objects and advantages of the invention will be apparent to those skilled in the art upon reading of the disclosure hereof.
In accordance with the invention, a synthetic control standard for the determination of cerebrospinal fluid is prepared from normal blood serum by dilution with a reagent comprising glucose and chloride ion in aqueous solution to a level whereby the total protein concentration ranges from about 30 to about 145 mg. percent. It is preferred that the diluted serum contains from about 40 to about 200 mg. percent glucose and from about to about 130 meq. per liter of chloride ion.
The chloride ion can be provided by use of an aqueous solution of an alkali metal chloride, preferably sodium chloride. The chloride salt and the glucose preferably are analytical or reagent grade materials.
For some purposes it is desired to include in the synthetic control standard of this invention a small amount of pre-albumin, preferably from about 1 to about 3 mg. percent. Pre-alburnin is a special fraction obtained from blood serum and is described, for example, by Got, et al., Isolement et caractrisation dune pralbumine du serum humain, Protides of the Biological Fluids, Vol. 10, H. Peeters, ed., Amsterdam, Elsevier, 1963, pp. 125-126; Schultze and Hermans, Molecular Biology of Human Protein, Vol. 1, Amsterdam, Elsevier, 1965, pp. l84-l85.
After dilution of the blood serum with aqueous glucose and saline or other chloride containing ion, with or without the added pre-albumin, the aqueous solution is lyophilized to a dry material. The lyophilized material is improved by the addition of a small amount of gum acacia prior to lyophilization, for example, on the order of about 0.1%. This improved lyophilized, dry material can be readily reconstituted with water prior to use without product loss on the container closure.
The synthetic cerebrospinal fluid control standard made as described herein has been found to produce much sharper bands in the electrophoretic pattern in actual use than obtained with the normal human cerebrospinal fluid. This is an added advantage of the invention in practice and enables the technician to improve the accuracy of the control procedures.
The following examples will further illustrate the invention, although the invention is not limited to these specific examples.
EXAMPLE 1 A synthetic control standard for use in the determination of cerebrospinal fluid is prepared as follows:
Human blood serum from which the clotting factors have been removed is diluted with an aqueous solution of glucose and sodium chloride containing 6.38 grams of NaCl and 600 mg. of glucose per liter. The serum is thereby diluted to a total protein concentration of 110 mg. per ml. by admixing 65 parts by volume of the diluent solution with one part by volume of the serum. To one liter of the diluted serum is then added 10 ml. of a 10% solution of gum acacia in water. The resulting solution is then filled in 3 ml. aliquots into 5 ml. bottles and lyophilized. The lyophilized, dry material is stored at 2 to 8 C. until required for use. In use, the dry material is reconstituted by dilution with 3 ml. of distilled water per bottle. Upon reconstitution of the foregoing material, the following assay values were determined.
Constituent: Value Chloride, meq. per liter 105 Glucose, mg. per 100 ml. 57 Protein, total, mg. per 100 ml. 110 Pre-albumin, percent of total Albumin, percent of total 58 a -globulin, percent of total 6 a -globulin, percent of total 12 fl-globulin, percent of total l3 -globulin, percent of total 11 Colloidal gold (Langes test) 1111100000 EXAMPLE 2 Example 1, above, is repeated except that 1-3 mg. percent of pre-albumin is added to the solution prior to lyophilization.
The control standards prepared in Examples 1 and 2, above, are used in the same manner as an unknown cerebrospinal fluid for comparison with the patients sample to provide a check on the analytical determination.
Various other examples and modifications of the foregoing examples can be made by the person skilled in the art after reading the foregoing description and the appended claims without departing from the spirit scope of the invention. All such further examples and modifications are included Within the scope of the appended claims.
What is claimed is:
1. The method of making a synthetic control standard for use in the determination of cerebrospinal fluid constituents comprising diluting normal blood serum with an aqueous diluent containing from about to about 200 mg. per 100 ml. of glucose and from about to about 130 milliequivalents per liter of chloride ion to a level at which the total protein concentration ranges from about 30 to about 145 mg. per ml.
2. The method of claim 1 including the additional steps of admixing the liquid control standard with about 0.1% gum acacia and lyophilizing to a dry, powdered material.
3. A synthetic control standard for use in the determination of cerebrospinal fluid comprising normal blood serum diluted with an aqueous diluent containing from about 40 to about 200 mg. per 100 ml. of glucose and from about 80 to about milliequivalents per liter of chloride ion to a level at which the total protein concentration ranges from about 30 to about mg. per 100 ml.
References Cited UNITED STATES PATENTS 3,627,468 12/1971 Goldenberg et al. 2323() B MAYER WEINBLA'IT, Primary Examiner U.S. Cl. X.R. 23-23O B; 424-2
US00237905A 1972-03-24 1972-03-24 Cerebrospinal fluid control standard Expired - Lifetime US3753925A (en)

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BE (1) BE797081A (en)
CA (1) CA985996A (en)
DE (1) DE2314263C2 (en)
FR (1) FR2185248A5 (en)
GB (1) GB1372812A (en)
ZA (1) ZA731898B (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3897363A (en) * 1973-08-10 1975-07-29 Baxter Laboratories Inc Blood control standard
US3920580A (en) * 1973-07-12 1975-11-18 Miles Lab Liquid control solution
US4127502A (en) * 1977-06-10 1978-11-28 Eastman Kodak Company Stabilizers for reconstituted, lyophilized samples
US4216117A (en) * 1978-05-15 1980-08-05 Bonderman Dean P Lipoprotein diluent or solution and method useful in the preparation of assay reference materials
US4663295A (en) * 1983-06-29 1987-05-05 Ciba Corning Diagnostics Corp. Estrogen-progesterone control reagents and methods for making same
WO1992015887A1 (en) * 1991-03-07 1992-09-17 Baxter Diagnostics Inc. Biosynthetic cerebrospinal fluid control and method of use

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3694322B1 (en) 2017-10-09 2024-07-03 Terumo BCT Biotechnologies, LLC Lyophilization container and method of using same
CA3130670A1 (en) 2019-03-14 2020-09-17 Terumo Bct Biotechnologies, Llc Lyophilization loading tray assembly and system

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3920580A (en) * 1973-07-12 1975-11-18 Miles Lab Liquid control solution
US3897363A (en) * 1973-08-10 1975-07-29 Baxter Laboratories Inc Blood control standard
US4127502A (en) * 1977-06-10 1978-11-28 Eastman Kodak Company Stabilizers for reconstituted, lyophilized samples
US4216117A (en) * 1978-05-15 1980-08-05 Bonderman Dean P Lipoprotein diluent or solution and method useful in the preparation of assay reference materials
US4663295A (en) * 1983-06-29 1987-05-05 Ciba Corning Diagnostics Corp. Estrogen-progesterone control reagents and methods for making same
WO1992015887A1 (en) * 1991-03-07 1992-09-17 Baxter Diagnostics Inc. Biosynthetic cerebrospinal fluid control and method of use
US5427949A (en) * 1991-03-07 1995-06-27 Dade International Inc. Biosynthetic cerebrospinal fluid control

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BE797081A (en) 1973-07-16
CA985996A (en) 1976-03-23
GB1372812A (en) 1974-11-06
JPS4931390A (en) 1974-03-20
FR2185248A5 (en) 1973-12-28
ZA731898B (en) 1973-12-19
DE2314263A1 (en) 1973-10-04
DE2314263C2 (en) 1982-11-25

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