Composition and method for hemoglobin and cell analysis

A technology of hemoglobin and hemoglobin concentration, which can be used in the analysis of materials, biomaterial analysis, biological testing, etc., and can solve problems such as lack and toxicity

Inactive Publication Date: 2004-06-16
COULTER INTERNATIONAL CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0018] As noted above, current hemoglobin measurement reagents and methods are either toxic or lack the ability to perform multiple diagnostic assays in the same automated step, such as hemoglobin determination and counts of white blood cells or differential counts of white blood cell subsets

Method used

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  • Composition and method for hemoglobin and cell analysis
  • Composition and method for hemoglobin and cell analysis
  • Composition and method for hemoglobin and cell analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0113] Prepare reagents of the following composition.

[0114] Recipe 1a

[0115] Tetrazole 5.0g

[0116] Tetradecyltrimethylammonium bromide 15.0g

[0117] Distilled water, adjusted to 1 L

[0118] pH 2.77

[0119] Recipe 1b

[0120] Triazole 10.0g

[0121] Dodecyltrimethylammonium chloride (50% solution) 35.0ml

[0122] Tetradecyltrimethylammonium bromide 3.5g

[0123] Distilled water, adjusted to 1 L

[0124] pH 6.19

[0125] Recipe 1c

[0126] Quinalidine Acid 5.0g

[0127] Dodecyltrimethylammonium chloride (50% solution) 50.0ml

[0128] Distilled water, adjusted to 1 L

[0129] pH 2.55

[0130] Recipe 1d

[0131] Melamine 5.0g

[0132] Tetradecyltrimethylammonium bromide 3.6g

[0133] Dodecyltrimethylammonium chloride (50% solution) 36.0ml

[0134] Distilled water, adjusted to 1 L

[0135] pH 4.55

[0136] Recipe 1e

[0137] Tetrazole 5.0g

[0138] Chemfac NB-104

[0139] (complex phosphate manufactured by Chemax Corporation) 30.0g

[0140] Distille...

Embodiment 2

[0144] Prepare reagents of the following composition.

[0145] Tetrazole 5.0g

[0146] Dodecyltrimethylammonium chloride (50% solution) 36.0ml

[0147] Tetradecyl ammonium bromide 3.6g

[0148] distilled water 1 liter

[0149] pH 2.73

[0150] 11.6 μl of whole blood samples were diluted with 2500 μl of blood thinner, then 403 μl of the above lysing agent composition were manually mixed with the pre-diluted samples. Absorption spectra of the samples were immediately measured on a Beekman DU7500 spectrophotometer. Five commercial blood thinners were used for hemoglobin determination, the COULTER  ISOTON  III.COULTER  ISOTON  II. Technicon H TM Systems RBC DIL (commercially available from Technicon Instruments), standard phosphate buffered saline and standard sodium chloride saline. The chromogen formed has an absorption maximum at about 540 nm and a shoulder at about 565 nm. When different diluents are used for the determination of hemoglobin, the maximum abso...

Embodiment 3

[0152] Prepare reagents of the following composition.

[0153] Recipe 3a

[0154] Tetrazole 5.0g

[0155] Tetradecyltrimethylammonium bromide 15.0g

[0156] Distilled water, adjusted to 1 L

[0157] pH 12.06

[0158] Recipe 3b

[0159] Aniline-2-sulfonic acid 5.0g

[0160] Tetradecyltrimethylammonium bromide 10.0g

[0161] Cetyltrimethylammonium bromide 4.0g

[0162] distilled water 1 liter

[0163] pH 1.67

[0164] Recipe 3c

[0165] Quinalidine Acid 5.0g

[0166] Dodecyltrimethylammonium chloride (50% solution) 40.0ml

[0167] distilled water 1 liter

[0168] pH 2.58

[0169] About 70 blood samples (with a hemoglobin concentration of 6-17g / dL and a white blood cell count of 1,000 / μL-40,000 / μL) were collected on a calibrated COULTER  On the STKS instrument under standard instrument settings (except solvent, LYSE S  III diff was replaced by the above preparation) analysis. For each formulation, approximately half of the samples were clinical samples for vario...

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Abstract

A cyanide-free lytic reagent composition and method for measuring the total hemoglobin concentration in a blood sample, for counting the number of leukocytes and for deferential counting of leukocyte subpopulations are described. The cyanide-free lytic reagent composition comprises a quaternary ammonium salt or a pyridinium salt to lyse erythrocytes and release hemoglobin, and an organic ligand selected from the group consisting of triazole and its derivatives, tetrazole and its derivatives, alkaline metal salts of oxonic acid, melamine, aniline-2-sulfonic acid, quinaldic acid, 2-amino-1,3,4-thiadiazole, triazine and its derivatives, urazole, DL-pipecolinic acid, isonicotinamide, anthranilonitrile, 6-aza-2-thiothymine, 3-(2-thienyl)acrylic acid, benzoic acid and alkali metal and ammonium salts of benzoic acid, and pyrazine and its derivatives to form a stable chromogen with hemoglobin, and a salt to adjust conductivity of the reagent for impedance measurement. The reagent composition is mixed with a blood sample without pre-dilution and the UV absorption of the sample mixture is measured at the predetermined absorption wavelength. Counting the number of leukocytes and differential counting of leukocyte subpopulations are accomplished simultaneously on an automated cell counter utilizing DC impedance measurement.

Description

[0001] related application [0002] This application is a continuation-in-part of co-pending US Application Serial No. 08 / 786,505, filed January 21,1997. Background of the invention [0003] The technical field of the invention [0004] The invention relates to a dissolving agent composition, a diluent and a method for manually or automatically measuring the total hemoglobin concentration in a blood sample and simultaneously performing leukocyte counting or differential counting of leukocyte subsets. [0005] discussion of prior art [0006] The measurement of total hemoglobin is an indicator of the oxygen-carrying capacity of whole blood. The ability to measure hemoglobin (Hgb) in a blood sample is a very important part of diagnostic assays and is also important in monitoring response to therapies for diseases that affect hemoglobin, as well as to therapies for other diseases that may adversely affect hemoglobin levels . [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/48G01N33/49G01N33/50G01N33/569G01N33/72
CPCG01N33/56972G01N33/721G01N33/5094Y10T436/10Y10T436/101666Y10T436/105831Y10T436/107497Y10T436/108331Y10T436/25125
Inventor 李毅C·杨
Owner COULTER INTERNATIONAL CORPORATION
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