GB2026686A - Clinical Procedure for Measuring Organic Lipid Molecules - Google Patents
Clinical Procedure for Measuring Organic Lipid Molecules Download PDFInfo
- Publication number
- GB2026686A GB2026686A GB7912870A GB7912870A GB2026686A GB 2026686 A GB2026686 A GB 2026686A GB 7912870 A GB7912870 A GB 7912870A GB 7912870 A GB7912870 A GB 7912870A GB 2026686 A GB2026686 A GB 2026686A
- Authority
- GB
- United Kingdom
- Prior art keywords
- concentration
- applying
- support medium
- lipoprotein
- triglyceride
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/04—Endocrine or metabolic disorders
- G01N2800/044—Hyperlipemia or hypolipemia, e.g. dyslipidaemia, obesity
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Wood Science & Technology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A method of determining the concentration in body fluid of different density fractions of organic lipid molecules comprising the steps of: a) applying a sample of the body fluid to be tested to a solid electrophoresis support medium, b) applying a direct current across the electrophoresis support medium for a predetermined period of time sufficient to separate the organic molecules into their separate density fractions, c) applying a developing substrate to the electrophoresed substrate sensitive to the organic lipid molecules, and d) quantitatively determining the concentrations of each of the separated density fractions of the organic lipid molecules from the developed electrophoresed sample. The quantitative measurement may be made by optical transmission or fluorescence or by radioactive tagging.
Description
SPECIFICATION
Clinical Procedure for Measuring Organic Lipid Molecules
The present invention relates to a clinical method of determining the concentration in body fluid of different density fractions of organic lipid molecules. In particular it relates to a clinical method of determining the concentration of lipoprotein free cholesterol fractions, particularly high density lipoprotein (HDL) free cholesterol, in serum, plasma and other body fluids. It also relates to a clinical method of determining the concentration of lipoprotein triglyceride fractions, particularly high density lipoprotein (HDL) triglycerides, in serum, plasma and other body fluids.
Our co-pending application No. 37413/78, (Serial No. 2704915) discloses and claims a method of determining the density fractions of the combination of free cholesterol and cholesterol esters in a body fluid sample. It is now believed that free cholesterols, particularly the high density lipoprotein fraction, is an even better indicator or predictor of cardiovascular risk. However, a reliable and simple method of determining the high density lipoprotein fraction of free cholesterols in body fluid has not been available.
Blood serum cholesterol has been recognized for over thirty years, and blood serum triglycerides for over ten years, as associated with coronary artery diseases. Medical experts have long believed that persons having elevated serum cholesterol levels or serum triglyceride levels are more likely to suffer myocardial infarcation (heart attack) than persons having lower levels of cholesterol or triglycerides.
However, the correlation between cholesterol or triglyceride levels and coronary artery disease is not consistent and therefore the present diagnostic test for cholesterol or triglyceride are considered advisory only and not a reliable indicator of the likelihood of myocardial infarcation or premature coronary artery disease.
The more recent work by the National Heart, Lung and Blood Institute of Bethesda, Maryland and the Framingham Heart Institute af Framingham, Massachusetts has suggested that one fraction of cholesterol, high density lipoprotein, is actually a "predictor of inverse cardiovascular risk". This discovery should improve our understanding of the role of cholesterol in coronary artery diseases.
Further, a correlation between free cholesterol and etherified cholesterol in each lipoprotein fraction (high density lipoprotein, low density lipoprotein and very low density lipoprotein HDL, LDL and VLDL) and cardiovascular risk, and a correlation between lipoprotein triglyceride fractions and cardiovascular risk may also be found. Therefore, a simple, fast and reliable test for the concentrations of lipoprotein free cholesterol fractions or lipoprotein triglyceride fractions in body fluids is necessary.
The present clinical tests for determining the concentration of high density lipoprotein (HDL) free cholesterol or triglyceride in body fluid requires precipitation of the other cholesterol or triglyceride fractions (low density and very low density lipoproteins) and determination of the free cholesterol or triglyceride concentration in the supernate. Briefly, the recommended test includes adding heparin solution to the fluid sample and mixing, adding manganese chloride and mixing, chilling and drawing off the supernate. All of the free cholesterol or the triglyceride remaining in the supernate is assumed to be high density lipoprotein free cholesterol or triglyceride. The cholesterol or triglyceride is extracted with isopropanol and the extract assayed for free cholesterol or triglyceride in spectrophotometers or continuous-flow analyzers.
It will be apparent that this procedure has several disadvantages. The procedure is slow and therefor expensive. Because precipitation is used, the reliability of the test is suspect. Finally, the assumption that all of the free cholesterol or triglyceride remaining after precipitation is high density lipoprotein has been seriously questioned. The problem with this assumption is the lack of specificity of the commonly used cations, specifically Ca2+, Mg2+ and Mn2+, in the lipoprotein-heparin interaction.
Further, it has been found that sub-classes of high density lipoproteins can be precipitated in the presence of manganese cations. Thus, the precipitation method may not be as reliable as believed.
The method of determining the concentration of lipoprotein free cholesterol fractions or lipoprotein triglyceride fractions of the present invention eliminates these problems and provides a simple and reliable clinical procedure.
Cholesterol occurs in blood serum in two forms namely free cholesterol and cholesterol esters.
Both forms are bound to serum proteins along with other lipids (e.g. triglycerides, phospholipids, ec.) to from lipoproteins.
Triglyceride also occurs in blood serum bound to serum proteins along with other lipids (e.g.
cholesterols, phospholipids, etc) to form lipoprotein. These lipoproteins occur in different densities as initially determined by ultracentrifugation. The density fractions are generally referred to as high density lipoproteins (HDL) cholesterol, very low density lipoprotein (VLDL) cholesterol and low density lipoproteins (LDL) cholesterols. It will be understood that further fractions have been identified including subclasses of HDL cholesterol, however these are the principal features. As set forth above, the present method provides a simple and reliable determination of the density fractions of free cholesterol in body fluids, providing an important additional indicator for clinical testing.
Although certain proteins have been separated by electrophoresis methods, such methods have not been successful in separating smaller molecules such as cholesterol or triglyceride. The method of this invention takes advantage of the fact that free cholesterol or triglycerides are bound to serum proteins, permitting separation by electrophoresis methods.
According to the present invention a method of determining the concentration in body fluid of
A method of determining the concentration in body fluid of different density fractions of organic lipid molecules comprising the steps of:
a) applying a sample of the body fluid to be tested to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium for a predetermined period of time sufficient to separate the organic lipid molecules into their separate density fractions,
c) applying a developing substrate to the electrophoresed substrate sensitive to concentrations of the organic lipid molecules, and
d) quantitatively determining the concentrations of each of the separated density fractions of the organic lipid molecules from the developed electrophoresed sample.
The invention includes a method of determining the concentration in body fluid of different density fractions of organic lipid molecules, other than lipoprotein cholesterols, comprising the steps of:
a) applying a sample of the body fluid to be tested to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium for a predetermined period of time sufficient to separate the organic lipid molecules into their separate density fractions,
c) applying a developing substrate to the electrophoresed substrate sensitive to the organic lipid molecules, and
d) quantitatively determining the concentrations of each of the separated density fractions of the organic lipid molecules from the developed electrophoresed sample.
The invention also includes a method of determining the concentration in body fluid of high density lipoprotein free cholesterol comprising:
a) applying a sample of the body fluid to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium until the high density lipoprotein free cholesterol has separated from any remaining lipoprotein in the sample,
c) applying a developing substrate sensitive to high density lipoprotein free cholesterols to the separate electrophoresed high density lipoprotein free cholesterols, and
d) quantitatively determining the concentration of the high density lipoprotein free cholesterol present in the body fluid from the developed electrophoresed sample.
The invention further includes a method of determining the concentration in body fluid of high density lipoprotein triglyceride comprising:
a) applying a sample of the body fluid to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium until the high density lipoprotein triglyceride has separated from any remaining lipoprotein in the sample,
c) applying a developing substrate sensitive to high density lipoprotein triglycerides to the separate electrophoresed high density lipoprotein triglyceride, and
d) quantitatively determining the concentration of the high density lipoprotein triglyceride present in the body fluid from the developed electrophoresed sample.
By means of this invention it is possible to determine the concentrations of high density, very low density and low density lipoprotein free cholesterol or high density, very low density and low density lipoprotein triglyceride. These are the preferred embodiments and the remainder of the description is concerned with the aspects of the invention.
The procedure is as follows:- First, a small sample of the body fluid to be tested is applied to a solid electrophoresis support medium, preferably cellulose acetate. The support medium will generally be in the form of a strip. Next, a direct current is applied across the support medium for a predetermined time to separate the high density, very low density and low density lipoprotein cholesterols or triglycerides on the medium. Next, a developing substrate sensitive to small concentrations of free cholesterol or triglyceride is applied to the electrophoresed lipoprotein free cholesterols or triglyceride, developing the separated lipoprotein free cholesterols or triglyceride on the support medium, the free cholesterols appearing reddish-brown in color and the triglycerides appearing reddish in color.Finally, the concentrations of each of the lipoprotein free cholesterols or the triglycerides may be quantitatively determined by one or any of several methods, including direct densitometry or by eluting each fraction and measuring the concentration of each lipoprotein in the eluate.
The method of the present invention thus permits substantially simultaneous measurement of high density lipoprotein free cholesterol, low density lipoprotein free cholesterol and very low density lipoprotein free cholesterol or high density lipoprotein triglyceride, low density lipoprotein triglyceride and very low density lipoprotein triglycerides. The procedure is faster and less costly than the present clinical methods because the procedure of the present invention eliminates precipitation. Finally, the method of the present invention is more reproducible because the determination of the lipoprotein free cholesterol or triglyceride concentrations are made directly from the entire sample. Other advantages and meritorious features of the present invention will be more fully understood from the following detailed description.
The method of determining concentrations of lipoprotein free cholesterols or trigiycerides of this invention is basically and electrophoretic determination. Thus, a small sample of the body fluid to be tested is first applied to a solid electrophoresis support medium, preferably cellulose acetate. A suitable cellulose acetate support medium is available in strip form from Helena Laboratories Corporation of
Texas, U.S.A. under the trade name "Titan III". It will be understood that other support media for example cellulose nitrate, agar, agarose, paper acrylamide gel, cellulose propionate, silica gel or starch gel, may be used. The fluid sample is preferably applied to the support medium in a straight line, permitting accurate reading following electrophoresis.A suitable apparatus for applying the fluid sample to the support medium is disclosed in United States Patent No. 4,006,705.
Next, a direct electric current is applied across the medium, causing separation of the lipoprotein free cholesterol or triglyceride fractions. Movement of the lipoprotein free cholesterols or triglycerides through a medium such as cellulose acetate depends upon the medium, the intensity of the electric field, the time and the character of the charged particle. In view of the fact that these variables will be constant for each lipoprotein free cholesterol or triglyceride fraction, the fractions are separated upon application of the electric field. It has been found that optimum separations for lipoprotein free cholesterols or triglycerides occur at about one hundred and eighty (180) volts (DC) for about twenty minutes. It has been found that the order of separation is HDL, VLDL and LDL free cholesterol or triglyceride, which is the order given herein.
Following electrophoresis, a developing substrate sensitive to small concentrations of cholesterol or triglyceride is applied to the electrophoresed lipoprotein cholesterol or triglyceride strip. In the preferred embodiment for the cholesterols, the developing substrate is a cholesterol oxidase substrate such as available from Boehringer Mannheim Corporation, Indianapolis, Indiana. The cholesterol oxidase available from Boehringer Mannheim Corporation is used to measure free cholesterol enzymatically.A suitable cholesterol oxidase substrate may have the following formulation:
Sodium cholate 58.0 mM
4-Aminoantipyrine 8.0 mM
Phenol 106.0 mM
Peroxidase (horseradish) 20.0 U/mL
Cholesterol oxidase
(bacterial) 0.75 U/mL
Buffer and other non
reactive ingredients
In the preferred embodiment for the triglycerides the developing substrate is an enzymatic triglyceride reagent such as available from Dow Diagnostics, Indianapolis, Indiana.The preferred enzymatic triglyceride reagent available from Dow Diangnostics has the following compositions:
Concentration Ingredient
0.3 mg Magnesium Chloride (MgC12) 1.8 mg Adenosine-5'
Triphosphate (ATP)
7.2 mg Nicotinamide adenine
dinucleotide (NAD)
0.25 mg 2-(p-iodophenyl-3-p-5
phenyltetrazolium
chloride (INT)
5.52 u(activity units)/test Diaphorase
0.92 u/test Glycerol Kinase 1 3.8 u/test Glycerol-i -phosphate
dehydrogenase
100 u/test Lipases
The lyphylized sample is diluted in 1 ml. of a buffer having a pH 7.6.
In the method of the present invention, the electrophoresced lipoprotein triglycerides are incubated with the triglyceride substrate for about fifteen minutes at 37C Centigrade.
The developing substrate may be applied to the electrophoresed free cholesterols or triglycerides by one of several methods, including simply soaking or submerging the support medium in the reagent or, more preferably, sandwiching the support medium to another support medium that has been impregnated with the reagent. For example, a strip of cellulose acetate as described above may be soaked or impregnated with cholesterol oxidase reagent or triglyceride reagent. A sandwich of a strip of cellulose acetate impregnated with the reagent and the electrophoresed medium is then made, which is incubated as described above.
Where the developing substrate is a cholesterol oxidase reagent, the lipoprotein free cholesterols are stained a reddish-brown colour and are easily visualized on the support medium. Where the developing substrate is an enzymatic triglyceride reagent the lipoprotein triglycerides are stained a reddish colour are also easily visualized on the support medium. Further, as described, the lipoprotein fractions have been separated during electrophoresis, permitting quantitative determination of the concentration of high density lipoprotein, very low density lipoprotein and low density lipoprotein free cholesterols or triglycerides.
Quantitation may be accomplished by one af several methods. In the simplest method, the support medium is scanned by a suitable instrument for measuring absorbence, such as a densitometer. Alternatively, the individual fractions may be eluted and the absorbance measured by a spectrophotometer. As will be understood by those skilled in the art, other quantitative methods may also be utilized. For example, the cholesterol oxidase reagent or the triglycerides may be tagged with fluorescein or a radioactive isotope, such as iodine 1 25. Where fluorescein is used, the concentration of each fraction may be determined by a fluorescent densitometry or spectrophotometry. Where a radioactive isotope is utilized, the concentrations are determined by measuring the radioactivity of each sample using a radioisotope scanner. Additionally, where a thin sheet or strip of cellulose acetate is used for the electrophoresis support medium, the individual lipoprotein fractions may be cut out with scissors. Then, each fraction may be dissolved and the fluorescence or radioactivity of each sample measured. This provides a very accurate determination.
It will be understood by those skilled in the art that various modifications may be made to the method of determining the concentration of lipoprotein free cholesterols or triglycerides of this invention. Further, details of the electrophoretic method will be understood by those skilled in the art,
For example, United States Patent No. 4,005,434 discloses a method and apparatus for graphic densitometer display which may be used in the method of this invention.
Claims (25)
1. A method of determining the concentration in body fluid of different density fractions of organic lipid molecules comprising the steps of:
a) applying a sample of the body fluid to be tested to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium for a predetermined period of time sufficient to separate the organic molecules into their separate density fractions,
c) applying a developing substrate to the electrophoresed substrate sensitive to the organic lipid molecules, and
d) quantitatively determining the concentrations of each of the separated density fractions of the organic lipid molecules from the developed electrophoresed sample.
2. A method of determining the concentration in body fluid of different density fractions of organic lipid molecules, other than lipoprotein cholesterols, comprising the steps of:
a) applying a sample of the body fluid to be tested to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium for a predetermined period of time sufficient to separate the organic lipid molecules into their separate density fractions,
c) applying a developing substrate to the electrophoresed substrate sensitive to the organic lipid molecules, and
d) quantitatively determining the concentrations of each of the separated density fractions of the organic lipid molecules from the developed electrophoresed sample.
3. A method of determining the concentration of high density lipoprotein free cholesterol in body fluid comprising: a) applying a sample of the body fluid to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium until the high density lipoprotein free cholesterol has separated from any remaining lipoprotein in the sample,
c) applying a developing substrate sensitive to high density lipoprotein free cholesterol to the separated electrophoresed high density lipoprotein free cholesterol, and
d) quantitatively determining the concentration of the high density lipropotein free cholesterol present in the body fluid from the developed electrophoresed sample.
4. A method as claimed in claim 3 wherein the developing substrate is a cholesterol oxidase substrate.
5. A method as claimed in claim 4 wherein the cholesterol oxidase is applied to the support medium by immersing the medium in a fluid sample of the cholesterol oxidase reagent.
6. A method as claimed in claim 4 wherein the cholesterol oxidase substrate is applied to the support medium by impregnating an untreated strip of support medium with fluid cholesterol oxidase reagent and applying the impregnated strip to the electrophoresed lipoprotein free cholesterols in a sandwich form and incubating the sandwiched medium for a predetermined period of time.
7. A method as claimed in any of claims 3 to 6 wherein the quantitative determination is made by a densitometer by measuring absorbence of each lipoprotein free cholesterol following application of the developing substrate.
8. A method as claimed in any of claims 3 to 6 wherein the quantitative determination is made by eluting the electrophoresed fraction and then quantitatively determining the concentration of the fraction.
9. A method as claimed in claim 8 wherein the concentration of the fraction is determined using a spectrophotometer.
1 0. A method as claimed in claim 8 wherein the cholesterol oxidase reagent is tagged with fluorescein including quantitatively determining the concentration of the fraction by measuring the fluorescence.
11. A method as claimed in claim 8 wherein the cholesterol oxidase is tagged with a radioactive isotope, including quantitatively determining the concentration of the fraction by measuring the radioactivity of the fraction with a radioisotope counter.
12. A method as claimed in any of claims 3 to 11 wherein the direct current separates high density, very low density and low density lipoprotein free cholesterols present in the fluid and wherein the concentration of each is quantitatively determined.
13. A method of determining the concentration of high density lipoprotein triglyceride in body fluid comprising: a) applying a sample of body fluid to a solid electrophoresis support medium,
b) applying a direct current across the electrophoresis support medium until the high density lipoprotein triglyceride has separated from any remaining lipoprotein in the sample,
c) applying a developing substrate sensitive to high density lipoprotein triglyceride to the separated electrophoresed high density lipoprotein triglyceride, and
d) quantitatively determining the concentration of the high density lipoprotein triglyceride present in the body fluid from the developed electrophoresed sample.
14. A method as claimed in claim 1 3 wherein the developing substrate is an enzymatic triglyceride reagent.
1 5. A method as claimed in claim 14 wherein the triglyceride reagent is applied to the support medium by immersing the medium in a fluid sample of the triglyceride reagent.
1 6. A method as claimed in claim 14 wherein the enzymatic triglyceride substrate is applied to the support medium by impregnating an untreated strip of support medium with fluid triglyceride reagent and applying the impregnated strip to the electrophoresed lipoprotein triglycerides in a sandwich form and incubating the sandwiched medium for a predetermined period of time.
17. A method as claimed in any of claims 1 3 to 1 6 wherein the quantitative determination is made by a densitometer by measuring absorbence of each lipoprotein triglyceride following application of the developing substrate.
18. A method as claimed in any of claims 13 to 1 6 wherein the quantitative determination is made by eluting the electrophoresed fraction, and then quantitatively determining the concentration of the fraction.
1 9. A method as claimed in claim 1 8 wherein the concentration of the fraction is determined using a spectrophotometer.
20. A method as claimed in claim 1 8 wherein the triglyceride reagent is tagged with fluorescein, including quantitatively determining the concentration of the fraction by measuring the fluorescence.
21. A method as claimed in claim 1 8 wherein the triglyceride reagent is tagged with a radioactive isotope, including quantitatively determining the concentration of the fraction by measuring the radioactivity of the fraction with a radioisotope counter.
22. A method as claimed in any of claims 13 to 21 wherein the direct current separates high density, very low density and low density lipoprotein triglycerides present in the fluid and wherein the concentration of each is quantitatively determined.
23. A method as claimed in any of the preceding claims wherein the electrophoresis support medium is cellulose acetate.
24. A method as claimed in any of the preceding claims wherein the direct current is about one hundred and eighty (180) volts which is applied to the support medium for about twenty minutes.
25. A method for determining the concentration in body fluids of different fractions of organic lipid molecules substantially as herein described.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US05/928,049 US4147606A (en) | 1977-09-21 | 1978-07-26 | Clinical procedure for measuring lipoprotein triglycerides |
| US05/928,044 US4167467A (en) | 1977-09-21 | 1978-07-26 | Clinical procedure for measuring lipoprotein free cholesterols |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| GB2026686A true GB2026686A (en) | 1980-02-06 |
Family
ID=27129953
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| GB7912870A Withdrawn GB2026686A (en) | 1978-07-26 | 1979-04-11 | Clinical Procedure for Measuring Organic Lipid Molecules |
Country Status (1)
| Country | Link |
|---|---|
| GB (1) | GB2026686A (en) |
-
1979
- 1979-04-11 GB GB7912870A patent/GB2026686A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4147606A (en) | Clinical procedure for measuring lipoprotein triglycerides | |
| Rosalki et al. | Two new methods for separating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma. | |
| Bronzert et al. | New micromethod for measuring cholesterol in plasma lipoprotein fractions. | |
| Beaumont et al. | Classification of hyperlipidaemias and hyperlipoproteinaemias. | |
| Legaz et al. | Electrophoretic amylase fractionation as an aid in diagnosis of pancreatic disease. | |
| Baena et al. | Analysis of carboxylic acids in biological fluids by capillary electrophoresis | |
| US4366244A (en) | Method for measuring serum cholesterol | |
| US4030995A (en) | Alkaline phosphatase isoenzyme determination | |
| US4167467A (en) | Clinical procedure for measuring lipoprotein free cholesterols | |
| BR0012311B1 (en) | method and agent for pretreatment of cholesterol quantification sample, method and kit for cholesterol quantification in specific lipoproteins using the same, free and total cholesterol quantification method. | |
| Okada et al. | Direct measurement of HDL cholesterol: method eliminating apolipoprotein E‐rich particles | |
| Weijers et al. | Partial characterization, properties, and clinical significance of a lactate dehydrogenase-immunoglobulin A kappa complex in serum. | |
| Schreiber et al. | Alkaline phosphatase isoenzymes resolved by electrophoresis on lectin-containing agarose gel. | |
| AU2003289081A1 (en) | Method of multiple quantification of cholesterol in low-density lipoproteins | |
| Fletcher et al. | A simple method for separating serum lipoproteins by electrophoresis on cellulose acetate | |
| Stuart et al. | Chloroplast cytochrome b6: molecular composition as a lipoprotein | |
| US6576106B1 (en) | Method for separating and assaying lipoprotein, an assembly for performing such a method, and a system including such an assembly | |
| Taylor et al. | Enzymatic and colorimetric determination of total serum cholesterol. An undergraduate biochemistry laboratory experiment | |
| GB2026686A (en) | Clinical Procedure for Measuring Organic Lipid Molecules | |
| CA2366663A1 (en) | Method for quantitating cholesterol | |
| EP0676642A1 (en) | Method for quantitatively analyzing a component in a lipoprotein fraction | |
| Karai et al. | Alterations of lipids of the erythrocyte membranes in workers exposed to lead | |
| Wong et al. | Comparison of 3 methods of analysis of the MB isoenzyme of creatine kinase in serum | |
| Demetriou et al. | Electrophoretic separation on agarose thin film of isoenzymes of alkaline phosphatase from human serum and tissue | |
| Jeevanandam et al. | A rapid, automated micromethod for measuring free fatty acids in plasma/serum. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| WAP | Application withdrawn, taken to be withdrawn or refused ** after publication under section 16(1) |