Classifications

• • 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
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/60—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving cholesterol
• • 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
  • G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
• • 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
  • Y10T436/104165—Lipid, cholesterol, or triglyceride 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
  • Y10T436/108331—Preservative, buffer, anticoagulant or diluent

Definitions

• the present invention relates to a cholesterol standard solution, more specifically, an aqueous solution of cholesterol with a definite and stable content of cholesterol.
• cholesterol solutions with a definite, known content of cholesterol as standard.
• cholesterol solutions are used with a definite cholesterol content which are called cholesterol standards.
• the cholesterol is dissolved in glacial acetic acid, chloroform, absolute alcohol or the like.
• the anhydrous standard solution so obtained gives, in the case of a cholesterol determination with the Liebermann- Burchardt color reagent, a color intensity which is different from that obtained with serum and can result in false cholesterol values in the serum (see L. Vass, Nurse. med. Wschr., 102 914/1972).
• aqueous cholesterol standard solution which contain hydroxypolyethoxydodecane as solubilizing agent, in turn suffer from a serious disadvantage insofar as the cholesterol content thereof changes rapidly and decreases noticeably even in the course of a few days.
• a cholesterol standard solution especially important that the cholesterol content thereof remains absolutely constant, it was hitherto necessary freshly to prepare the aqueous cholesterol standard solution before use, which was a considerable disadvantage and, in addition, rendered questionable the dependability of the cholesterol determination.
• the present invention obviates or mitigates these disadvantages and provides a stable, aqueous cholesterol standard solution.
• the present invention provides an aqueous cholesterol standard solution, comprising, in aqueous solution, a known amount of cholesterol, together with 1 to 20 volume percent of a primary or secondary aliphatic alcohol containing up to 4 carbon atoms (for example, ethanol, isopropanol or n-butanol); 5 to 20 volume percent of hydroxy-polyethoxydodecane or 40 to 60 volume percent diethyleneglycol monobutyl ether; and 0.5 to 5 weight percent sodium chloride and/or 0.01 to 0.5 weight percent alkyl dimethyl-benzyl ammonium chloride with 8 to 18 carbon atoms in the alkyl radicals.
• a primary or secondary aliphatic alcohol containing up to 4 carbon atoms
• hydroxy-polyethoxydodecane or 40 to 60 volume percent diethyleneglycol monobutyl ether for example, ethanol, isopropanol or n-butanol
• the present invention rests on the surprising discovery that a solution of cholesterol in dilute aqueous alcohol which, as solubilizing agent, contains the given amount of hydroxy-polyethoxydodecane or diethyleneglycol monobutyl ether, and, as stabilizing agent, sodium chloride and/or alkyl dimethylbenzyl ammonium chloride, is outstandingly stable and, even when stored for a year or more under normal conditions, does not change with regard to the cholesterol content.
• a preferred cholesterol standard solution according to the present invention contains the dilute alcohol, hydroxy-polyethoxydodecane and sodium chloride. in this preferred composition, the best results have been obtained with a content of 8 to l2 vol. percent of an alcohol, for example ethanol, isopropanol or n-butanol, and 0.8 to L0 wt. percent sodium chloride. If, instead of sodium chloride, alkyl dimethyl-benzyl ammonium chloride is used, then the preferred amount is 0.05 to 0.15 wt. percent.
• Another preferred cholesterol standard solution according to the present invention comprises 8 to 12 vol. percent of an alcohol, 55 to 60 vol. percent diethylene glycol monobutyl ether and 0.05 to 0.15 wt. percent alkyl dimethyl-benzyl ammonium chloride.
• Yet another preferred cholesterol standard solution according to the present invention additionally contains a small amount of tetramethylurea.
• the amount thereof can be 0.5 to 5 vol. percent and is preferably 0.8 to 2.5 vol. percent.
• an alkali metal azide it can be expedient, for the prevention of bacterial attack, to add an alkali metal azide.
• the pH value must be adjusted to more than 6.5 by the addition of an appropriate buffer. It is preferred to add 0.05 to 0.2 percent and more, preferably about 0.1 percent of an alkali metal azide, for example sodium azide, in a buffer of pH 6.5 to 8, preferably of 7 to 8, for example, a triethanolamine buffer.
• the cholesterol standard solution according to the present invention can contain up to 400 mg cholesterol in 100 ml of the solvent mixture.
• the extraordinary storage stability of the solutions necessitates neither the exclusion of light nor the exclusion of oxygen. This is all the more surprising since, as is known, cholesterol, even in a solid state, must be stored under an inert gas and with the exclusion of light.
• the stabilizing effect clearly depends upon the interaction of the various components of the standard solution according to the present invention since, with the individual components alone, a stabilizing effect cannot be achieved. For example, when a solution which originally contains 400 mg cholesterol in 100 ml of a mixture of 10 vol. percent hydroxy-polyethoxydodecane and vol.
• EXAMPLE 1 40 g recrystallized cholesterol were dissolved in one liter hydroxypolyethoxydodecane by heating in a water-bath at 40C. This solution was diluted with doubly distilled water with a temperature of 40C, then ml of a 10 percent aqueous solution of alkyl dimethylbenzyl ammonium chloride and 1 liter ethanol were added thereto and, after cooling to ambient temperature, made up to precisely 10 liters with doubly distilled water.
• EXAMPLE 2 40 g cholesterol were dissolved in 1 liter hydroxy-polyethoxydodecane and 1 liter ethanol and thereafter made up to liters with a 0.9 percent aqueous solution of sodium chloride.
• EXAMPLE 3 40 g cholesterol were dissolved in 1 liter hydroxy-polyethoxydodecane, 1 liter n-butanol, 100ml of a 10 percent aqueous solution of alkyl dimethylbenzyl-ammonium chloride and 100 ml tetramethylurea and then made up to ID liters with doubly distilled water.
• EXAMPLE 4 40 g cholesterol were dissolved in 6 liters diethyleneglycol monobutyl ether, 0.5 liters ethanol and 100 ml of a 10 percent aqueous solution of alkyl dimethylbenzyl-ammonium chloride were added thereto and then the mixture was made up to 10 liters with doubly distilled water.
• Cholesterol standard solution comprising in aqueous solution a. a defined amount of cholesterol;
• component (c) is diethyleneglycol monobutyl ether.
• component (d) is a combination of sodium chloride and alkyl dimethyl benzyl ammonium chloride.
• component (b) is contained in an amount of from 8 to l2 volume percent of said solution.
• component (d) is alkyl dimethyl benzyl ammonium chloride and is contained in an amount of from 0.05 to 0.15 weight percent.
• Standard solution as claimed in claim 7 comprising 8 to 12 volume percent of said alcohol as component (b).
• Standard solution as claimed in claim 1 comprising 55 to 60 volume percent of diethylene glycol monobutyl ether as component (c) and 0.05 to 0. l 5 weight percent of alkyl dimethyl benzyl ammonium chloride as component (d).
• Standard solution as claimed in claim 1 additionally containing tetramethylurea.
• Standard solution as claimed in claim 14 wherein the content of said tetramethylurea is from 0.5 to 5 volume percent based on said standard solution.
• Standard solution as claimed in claim 15 wherein the content of said tetramethylurea is from 0.8 to L5 volume percent based on said standard solution.
• Standard solution as claimed in claim 1 also con- 7 taining an alkali metal azide and wherein said solution is buffered to a pH of at least 6.5.

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• Life Sciences & Earth Sciences (AREA)
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• Physics & Mathematics (AREA)
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• Biotechnology (AREA)
• General Health & Medical Sciences (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Biophysics (AREA)
• General Physics & Mathematics (AREA)
• Medicinal Chemistry (AREA)
• Food Science & Technology (AREA)
• Cell Biology (AREA)
• Pathology (AREA)
• General Engineering & Computer Science (AREA)
• Genetics & Genomics (AREA)
• Investigating Or Analysing Biological Materials (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
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Cited By (12)

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Citations (3)

• 1973
  • 1973-05-14 DE DE2324386A patent/DE2324386C2/de not_active Expired
• 1974
  • 1974-04-02 AT AT273574A patent/AT330368B/de not_active IP Right Cessation
  • 1974-04-10 AR AR253266A patent/AR198369A1/es active
  • 1974-04-30 US US465640A patent/US3891573A/en not_active Expired - Lifetime
  • 1974-04-30 IT IT22151/74A patent/IT1010304B/it active
  • 1974-05-03 NL NL7405961.A patent/NL159193B/xx not_active IP Right Cessation
  • 1974-05-09 CH CH631174A patent/CH592309A5/xx not_active IP Right Cessation
  • 1974-05-10 SE SE7406271A patent/SE393191B/xx unknown
  • 1974-05-10 GB GB2075274A patent/GB1423113A/en not_active Expired
  • 1974-05-13 CA CA199,919A patent/CA1016049A/en not_active Expired
  • 1974-05-13 DD DD178475A patent/DD110949A5/xx unknown
  • 1974-05-14 FR FR7416677A patent/FR2229966B1/fr not_active Expired
  • 1974-05-14 JP JP5378174A patent/JPS541197B2/ja not_active Expired
  • 1974-05-14 HU HUBO1499A patent/HU169654B/hu unknown

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