CA1088851A - Reference material for establishing anion concentrations in analytical chemistry tests - Google Patents
Reference material for establishing anion concentrations in analytical chemistry testsInfo
- Publication number
- CA1088851A CA1088851A CA278,995A CA278995A CA1088851A CA 1088851 A CA1088851 A CA 1088851A CA 278995 A CA278995 A CA 278995A CA 1088851 A CA1088851 A CA 1088851A
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- CA
- Canada
- Prior art keywords
- bicarbonate
- composition
- quaternary ammonium
- ammonium carbonate
- water
- Prior art date
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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
- 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
- G01N2496/00—Reference solutions for assays of biological material
- G01N2496/70—Blood gas control solutios containing dissolved oxygen, bicarbonate and the like
-
- 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
- G01N2496/80—Multi-analyte reference solutions containing cholesterol, glucose and the like
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- Urology & Nephrology (AREA)
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- Food Science & Technology (AREA)
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- 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)
Abstract
ABSTRACT OF THE DISCLOSURE
A composition for reconstituting blood serum to provide a control standard used in the analysis of blood serum, said composition comprising a quaternary ammonium carbonate or bicarbonate.
A composition for reconstituting blood serum to provide a control standard used in the analysis of blood serum, said composition comprising a quaternary ammonium carbonate or bicarbonate.
Description
This invention relates to ~1 laboratoxy standard material, and more particularly, to a 5 table blood serum re~er~nce standard and -to a method of prepara-tion and use -thereof.
Various types of apparatus have been developed in recent years for the au-toma-tic analysis of human blood serum, which apparatus are capable of making rapid simultaneous analytical determinations on a precise quantitative basis o~
various components of each of a series of blood serum samples successively introduced into the apparatus. Biochemical determinations may be made, for example, by such apparatus, of the following constituents of the blood serum samples; albumin, alkaline phosphatase, bilirubin, calcium, chloride, cholesterol, carbon dioxide content, creatinine, glucose, lactic dehydrogenase, inorganic phosphorus, potassium, sodium, -total protein, transaminase, urea nitrogen, uric acid and creatine phosphokinase.
In order to calibrate the apparatus of the type for making these determinations, a reference standard blood serum sample is used whose constituents have precisely predetermined values. It is necessary for such reference standard serums to be capable of being stored for long periods of time without deterioration, and for that reason, it has been the practice to freeze-dry or lyophilize them. It has also been the prior art practice to reconstitute the reference standard serum before use by means of, Eor example, a separate aqueous solution containing ammonium bicarbonate to restore the carbon dioxide that has been lost in the blood serum during the lyophilizing process. Examples of
Various types of apparatus have been developed in recent years for the au-toma-tic analysis of human blood serum, which apparatus are capable of making rapid simultaneous analytical determinations on a precise quantitative basis o~
various components of each of a series of blood serum samples successively introduced into the apparatus. Biochemical determinations may be made, for example, by such apparatus, of the following constituents of the blood serum samples; albumin, alkaline phosphatase, bilirubin, calcium, chloride, cholesterol, carbon dioxide content, creatinine, glucose, lactic dehydrogenase, inorganic phosphorus, potassium, sodium, -total protein, transaminase, urea nitrogen, uric acid and creatine phosphokinase.
In order to calibrate the apparatus of the type for making these determinations, a reference standard blood serum sample is used whose constituents have precisely predetermined values. It is necessary for such reference standard serums to be capable of being stored for long periods of time without deterioration, and for that reason, it has been the practice to freeze-dry or lyophilize them. It has also been the prior art practice to reconstitute the reference standard serum before use by means of, Eor example, a separate aqueous solution containing ammonium bicarbonate to restore the carbon dioxide that has been lost in the blood serum during the lyophilizing process. Examples of
-2--... . . . ~ ~ . . .
s~L
reconstituting solutions that have been used in the prior art can be found in U.S, P~t~nts Nos. 31~66,249 and 3,629,142.
Although the composition of this invention has as its primary purpose the reconstitution of lyophilized blood serum to return -to the serum a known quantity of carbona-te or bicarbonate so that the apparatus can be calibrated for determining the amount of carbon dioxide in the blood, the composition of this invention is useful for various other chemical analyses. As is well known, secondary reference materials serve two important functions in analytical chemistry laboratories. They are used as standards for calibrating instruments, and they are used to check the accuracy and precision of the assay results. These secondary reference materials usually contain multiple substances, so that just one solution can be used for most of the assays done by the laboratory.
Moreover, multiple-substance quality control reference materials are essential for use with multiple assay chemistry analysers.
In compounding these materials, there is often difficulty in fixing the concentration of-one or more anions at a desired level in the material without having the obligatory cations affect other laboratory assays. One very important example of the problem is that set forth above, namely, fixing the total carbon dioxide content (bicarbonate plus carbonate) in quality control or reference serums used in clinical chemistry laboratories.
.,.~, . , .. . , .. .. . -:., . ' , . : .. . . ..
sicarbonate s~lts wllich h~ve been used to ix totalcarbon diGxide content in these serums are -the bicarbona-tes o~ sodium, potassium, lithium, rubi~ium, TRIS (tris(hydroxymethyl) aminomethane) and ammonia. Sodium, potassium and lithium bicarbonates are unsuitable for the purposes described because sodium, potassium and lithiu, are important analytes, and because lithium is also used as an internal standard in the common flame photometric determination of sodium and potassium. Rubidium bicarbonate (at a concentration of 25mM) is unsuitable as a total carbon dioxide reference material because it elevates sodium determinations by about 2mM, possibly through a matrix effect in the flame photometer.
It has been found that the TRIS bicarbonate-based reference material disclosed in aforementioned U.S. Patent No. 3,629,142 has a major shortcoming for reference purposes because it depresses the urea result (urease-Berthelot method) by about 15 per cent, and enhances protein results by about 10 per cent. The effect of TRIS on the urea results may be related to interference with the serthelot reaction by a number of nitrogen-containing ~o compounds, as reported by Gips and Reitsema, "Clin. Chem. Acta.", 33, 257 (1971). Ammonium bicarbonate-reconstituted reference materials, which are the most widely used total carbon dioxide reference materials, are useless ~or most unreasebased procedures :~or determining urea, because of ammonium ion interference. The ammonium ion also severely disturbs specific electrodes employed ~or de-termininy potassium.
, ,~
Thus~ the presently available total carbon dioxide reference materials are unsuitable as reference materials for several other important analytical chemistry assays. A labora-tory performin~ total carbon dioxi~e measurements in addition to the assays which are interferred with by the above-mentioned total carbon dioxide reference materials must prepare two different lyophilized serum references, tha-t is, one reconstituted with water and the other reconstituted with a bicarbonate solution.
This duplication is burdensome, especially if a single multiple-channel instrument is performing the above tests, since twicethe usual number of standards an~/or quality controls are required.
We have found that duplication is eliminated and that expenses are reduced when the reference materials of -this invention are used as a total carbon dioxide reference, because the reference materials of the invention affect the level of no analyte of clinical chemical interest, save total carbon dioxide.
An additional advantage of the invention is the direct comparabil-ity of data for different laboratories participating in a regional quality control program (in which the participating laboratories all use the same lyophilized serum pool), even if some of the laboratories use water-reconstituted instead of bicarbonate-reconstituted serum controls.
This invention comprises the use of a salt selected from the class of compounds described below, for use in fixing the concentration o~ an anion in analytical chemistry reference materials, and which, when used for this purpose, gives no other ., .
interference with other cllemic~] ~ssays. The m~terials o~ thi5 invention, which we have used to s-tandardize and monitor -the quality of clinical chemistry assays, contain salts of quaternary ammonium compounds. Such compounds contain at least one tetrasubstituted nitrogen wherein certain of the substituent groups may be joined to form he-terocycles, for example, 1-alkylpyridine and 1, l-dialkylpiperidine, or polymerized via linkage of the substituent groups. ~he subskituent groups should exhibit minimal interference in clinical determinations-such as those set forth hereinafter, and the compounds themselves should be essentially free of contaminants, e.g., sodium, potassium, chloride, calcium, iron and ammonium. The amounts of these ions respectively which could be present in a commercially practicable diluent are no more than 2 millimolar, 0.25 millimolar, 3.0 millimolar, 0.5 milligrams/100 milliliters, 3.0 micrograms/100 milliliters and 3.0 micrograms/100 milliliters, with the preferred values being, respectively, 1.0 millimolar, 0.2 millimolar, 2.0 millimolar, 0.2 milligrams/100 ml, 2.0 micrograms/100 milliliters and 2.0 micrograms/100 milliliters, which would interfere in such determinations.
Additionally, the substituents should not render the compounds so insoluble that particles thereof would disrupt optical measurement of the reference ma-terial. Mos-t importantly, no substituents should in-troduce significant buffering capacity into the compounds. Exemplary substituent groups are the saturated or unsaturated heterocycles such as furan or tetrahydrothiophen radicals, cycloalkenyl groups such as phenol, OA ~
.
~ rj~
cycloalkyl groups, such as cyclohexyl and cyclopropyl, and branch chain or normal alkenyls including methyl, ethyl, propyl, butyl, pentyl, octyl, hexene, isobutyl and neohexyl. Further, any of the foregoing groups can be adclitionally substituted with or contain oxygen or thioe-thers, ketones, esters, halides, nitro or hydroxyl~
However, i-t is preferred to employ the brahch chain or normal alkyls or alkenyls as these are the most chemically inert substit-uent groups, with the most preferred being methyl. Each of the four substituent groups can be the same as or different from one or more of the remaining groups.
The anions which may be employed to form the quaternary ammonium salts may be, for example, carbonate, bicarbonate, mixtures of carbonate and bicarbonate, halide and any other anion of clinical interest. The preferred anion i5 carbonate or bicarbonate.
Using the preferred tetrasubstituted ammonium bicarbonate or carbonate salts of the present invention, the total carbon dioxide level in a serum or other biological fluid control may be conveniently adjusted to any desired level, without interfering with other chemistry procedures. Assays in which no interferences were found include the following assays: sodium, potassium, chloride, glucose, blood urea nitrogen, creatinine, calcium, phosphorus, total protein, alkaline phosphatase, total bilirubin, iron, tot~l iron binding capacity, cholesterol, triglycerides, uric acid, alanine aminotransferase, lactate dehydrogenase, hydroxybutyrate dehydrogenase, gamma-glutamyltransferase, lactic acid dehydrogenase, acid phosphatase, alkaline phosphatase, .
, - ~.O~
thyroid hormone uptake, total th~roid hormone and amylase. The bicarbonate materials add no color, so -that they do not affect colorimetric determinations. They do not cause precipi-tation of any of the other components oE the control material which could result in the plugging of instruments. In addition, the pH of the control solution can be adjusted by varying the ratio of bicarbonate to carbonate in the te-trasubstituted ammonium salt.
~n important feature of the present invention is that the tetra-substituted ammonium ion has no buffering capacity, and as a result, only minimal amounts of such ions are required in fixing the carbon dioxide content of the reference material, regardless of the pH. TRIS, on the other hand, is a buffer at pH 8, and therefore TRIS bicarbonate solutions contain more TRIS
than bicarbonate, since some of the TRIS is uncharged. Further, the presence of buffering capacity will interfere with titrimetric methods for determining CO2.
By way of example of the application of the present invention, the synthesis and use of tetramethylammonium bicarbonate (TMA-HCO3) as a total carbon dioxide content reference material is described below (see also the reaction scheme~. A solution of ~nalytically pure tetramethylammonium bromide of the desired concentration (for example, 0.15 M) was converted to tetramethylammonium hydroxide by passage through a chromatographic column containing a strongly basic exchange resin in the hydroxide form (step 1).
;~ -.
:, : . . .
- - . : ' ~ ' : ~ :: ' s~
0.15M ~C~13)~N BR S~p 0-15M (C~13)~sN 0~1 P
0,15M (C~13) ~N HC03 Conversion of the tetramethylammonium hydroxide to TMA-HC03 was accomplished by bubbling a s~ream of C02 gas into the stirred solution until a pH of 8 was obtained. The pH o~ the solution can be changed by changing the amount of C02 added to the solution. The TMA-HC03 solution was diluted to the desired total carbon dioxide content (for example 30mM). This Tl~A-HC03 solution was then used to reconstitute previously lyophilized serum controls. Alternatively, ~he TMA bicarbonate or carbonate salt may be added to the lyophilized serum control prior to its reconstitution with distilled water.
These TMA-CH03-reconstituted controls were then analyzed by known analytical chemistry procedures. Many such control serums so reconstituted were analyzed in parallel with controls reconstituted with only distilled water. No significant differences between these two controls was found for any of the assays in which the controls were used, which included those mentioned above.
In another application oE our invention, we have found that other te-trasubstituted ammonium anion salts can be used in analytical chemical laboratories. For example, tetrasubsti-tuted ammonium halide salts can be used as standards and controls in both clinical and toxicology laboratories. Like the tetrasubs-tituted ammonium bicarbonate and carbonate salts, these saits add no color to reference materials to which they are added, _9_ :: . - -. . .. , . .: . . ~- . : . : ..... , : . - ~ . .~ . , ,:~ .: . :. - . ..... , : : . ~ , . . ; : . . : .
:~ : : . .: :,.. : , . .. . . . . .. .. . ....
so that they do not affect colorimetric determinations. 'l'hey are chemically unreactive in other laboratory methods, and they do not cause precipitation of any of the other components of the control materials to which they may be added, which could result in the plugging of instrumen ts.
Tetrabutyl ammonium bromide-reconstituted control serum is an application of our invention which may be used to fix the 10 level of bromide in bromide assays. A solution of 30 mM
tetrabutylammonium bromide was used to reconstitute lyophilized control serum, in a manner analogous to that used for the tetraalkylammonium bicarbonates. The resulting tetrabutylammonium bromide control material was the~ analyzed by known analytical chemistry procedures. These control serums were analyzed in parallel with controls reconstituted with distilled water only.
No significant differences between these two controls was found for any of the assays in which the controls were ~used, except for the halide assays, in which the tetrabutylammonium bromide-20 reconstituted controls were elevated by an amount correspondingto 30 mM bromide.
The tetrasubstituted ammonium anion salts of this invention are known in the art. However, these salts have not been used in the analytical chemistry procedures in which the salts of this invention are being used~ The salts and their methods of produckion are disclosed in Czechoslovakian Patent No. 157,000 and in the U.S. Patents Nos. 3,190,919, 3,579,581, 3,539j605 and .," ' ' :
_, :~
IO~
s~L
reconstituting solutions that have been used in the prior art can be found in U.S, P~t~nts Nos. 31~66,249 and 3,629,142.
Although the composition of this invention has as its primary purpose the reconstitution of lyophilized blood serum to return -to the serum a known quantity of carbona-te or bicarbonate so that the apparatus can be calibrated for determining the amount of carbon dioxide in the blood, the composition of this invention is useful for various other chemical analyses. As is well known, secondary reference materials serve two important functions in analytical chemistry laboratories. They are used as standards for calibrating instruments, and they are used to check the accuracy and precision of the assay results. These secondary reference materials usually contain multiple substances, so that just one solution can be used for most of the assays done by the laboratory.
Moreover, multiple-substance quality control reference materials are essential for use with multiple assay chemistry analysers.
In compounding these materials, there is often difficulty in fixing the concentration of-one or more anions at a desired level in the material without having the obligatory cations affect other laboratory assays. One very important example of the problem is that set forth above, namely, fixing the total carbon dioxide content (bicarbonate plus carbonate) in quality control or reference serums used in clinical chemistry laboratories.
.,.~, . , .. . , .. .. . -:., . ' , . : .. . . ..
sicarbonate s~lts wllich h~ve been used to ix totalcarbon diGxide content in these serums are -the bicarbona-tes o~ sodium, potassium, lithium, rubi~ium, TRIS (tris(hydroxymethyl) aminomethane) and ammonia. Sodium, potassium and lithium bicarbonates are unsuitable for the purposes described because sodium, potassium and lithiu, are important analytes, and because lithium is also used as an internal standard in the common flame photometric determination of sodium and potassium. Rubidium bicarbonate (at a concentration of 25mM) is unsuitable as a total carbon dioxide reference material because it elevates sodium determinations by about 2mM, possibly through a matrix effect in the flame photometer.
It has been found that the TRIS bicarbonate-based reference material disclosed in aforementioned U.S. Patent No. 3,629,142 has a major shortcoming for reference purposes because it depresses the urea result (urease-Berthelot method) by about 15 per cent, and enhances protein results by about 10 per cent. The effect of TRIS on the urea results may be related to interference with the serthelot reaction by a number of nitrogen-containing ~o compounds, as reported by Gips and Reitsema, "Clin. Chem. Acta.", 33, 257 (1971). Ammonium bicarbonate-reconstituted reference materials, which are the most widely used total carbon dioxide reference materials, are useless ~or most unreasebased procedures :~or determining urea, because of ammonium ion interference. The ammonium ion also severely disturbs specific electrodes employed ~or de-termininy potassium.
, ,~
Thus~ the presently available total carbon dioxide reference materials are unsuitable as reference materials for several other important analytical chemistry assays. A labora-tory performin~ total carbon dioxi~e measurements in addition to the assays which are interferred with by the above-mentioned total carbon dioxide reference materials must prepare two different lyophilized serum references, tha-t is, one reconstituted with water and the other reconstituted with a bicarbonate solution.
This duplication is burdensome, especially if a single multiple-channel instrument is performing the above tests, since twicethe usual number of standards an~/or quality controls are required.
We have found that duplication is eliminated and that expenses are reduced when the reference materials of -this invention are used as a total carbon dioxide reference, because the reference materials of the invention affect the level of no analyte of clinical chemical interest, save total carbon dioxide.
An additional advantage of the invention is the direct comparabil-ity of data for different laboratories participating in a regional quality control program (in which the participating laboratories all use the same lyophilized serum pool), even if some of the laboratories use water-reconstituted instead of bicarbonate-reconstituted serum controls.
This invention comprises the use of a salt selected from the class of compounds described below, for use in fixing the concentration o~ an anion in analytical chemistry reference materials, and which, when used for this purpose, gives no other ., .
interference with other cllemic~] ~ssays. The m~terials o~ thi5 invention, which we have used to s-tandardize and monitor -the quality of clinical chemistry assays, contain salts of quaternary ammonium compounds. Such compounds contain at least one tetrasubstituted nitrogen wherein certain of the substituent groups may be joined to form he-terocycles, for example, 1-alkylpyridine and 1, l-dialkylpiperidine, or polymerized via linkage of the substituent groups. ~he subskituent groups should exhibit minimal interference in clinical determinations-such as those set forth hereinafter, and the compounds themselves should be essentially free of contaminants, e.g., sodium, potassium, chloride, calcium, iron and ammonium. The amounts of these ions respectively which could be present in a commercially practicable diluent are no more than 2 millimolar, 0.25 millimolar, 3.0 millimolar, 0.5 milligrams/100 milliliters, 3.0 micrograms/100 milliliters and 3.0 micrograms/100 milliliters, with the preferred values being, respectively, 1.0 millimolar, 0.2 millimolar, 2.0 millimolar, 0.2 milligrams/100 ml, 2.0 micrograms/100 milliliters and 2.0 micrograms/100 milliliters, which would interfere in such determinations.
Additionally, the substituents should not render the compounds so insoluble that particles thereof would disrupt optical measurement of the reference ma-terial. Mos-t importantly, no substituents should in-troduce significant buffering capacity into the compounds. Exemplary substituent groups are the saturated or unsaturated heterocycles such as furan or tetrahydrothiophen radicals, cycloalkenyl groups such as phenol, OA ~
.
~ rj~
cycloalkyl groups, such as cyclohexyl and cyclopropyl, and branch chain or normal alkenyls including methyl, ethyl, propyl, butyl, pentyl, octyl, hexene, isobutyl and neohexyl. Further, any of the foregoing groups can be adclitionally substituted with or contain oxygen or thioe-thers, ketones, esters, halides, nitro or hydroxyl~
However, i-t is preferred to employ the brahch chain or normal alkyls or alkenyls as these are the most chemically inert substit-uent groups, with the most preferred being methyl. Each of the four substituent groups can be the same as or different from one or more of the remaining groups.
The anions which may be employed to form the quaternary ammonium salts may be, for example, carbonate, bicarbonate, mixtures of carbonate and bicarbonate, halide and any other anion of clinical interest. The preferred anion i5 carbonate or bicarbonate.
Using the preferred tetrasubstituted ammonium bicarbonate or carbonate salts of the present invention, the total carbon dioxide level in a serum or other biological fluid control may be conveniently adjusted to any desired level, without interfering with other chemistry procedures. Assays in which no interferences were found include the following assays: sodium, potassium, chloride, glucose, blood urea nitrogen, creatinine, calcium, phosphorus, total protein, alkaline phosphatase, total bilirubin, iron, tot~l iron binding capacity, cholesterol, triglycerides, uric acid, alanine aminotransferase, lactate dehydrogenase, hydroxybutyrate dehydrogenase, gamma-glutamyltransferase, lactic acid dehydrogenase, acid phosphatase, alkaline phosphatase, .
, - ~.O~
thyroid hormone uptake, total th~roid hormone and amylase. The bicarbonate materials add no color, so -that they do not affect colorimetric determinations. They do not cause precipi-tation of any of the other components oE the control material which could result in the plugging of instruments. In addition, the pH of the control solution can be adjusted by varying the ratio of bicarbonate to carbonate in the te-trasubstituted ammonium salt.
~n important feature of the present invention is that the tetra-substituted ammonium ion has no buffering capacity, and as a result, only minimal amounts of such ions are required in fixing the carbon dioxide content of the reference material, regardless of the pH. TRIS, on the other hand, is a buffer at pH 8, and therefore TRIS bicarbonate solutions contain more TRIS
than bicarbonate, since some of the TRIS is uncharged. Further, the presence of buffering capacity will interfere with titrimetric methods for determining CO2.
By way of example of the application of the present invention, the synthesis and use of tetramethylammonium bicarbonate (TMA-HCO3) as a total carbon dioxide content reference material is described below (see also the reaction scheme~. A solution of ~nalytically pure tetramethylammonium bromide of the desired concentration (for example, 0.15 M) was converted to tetramethylammonium hydroxide by passage through a chromatographic column containing a strongly basic exchange resin in the hydroxide form (step 1).
;~ -.
:, : . . .
- - . : ' ~ ' : ~ :: ' s~
0.15M ~C~13)~N BR S~p 0-15M (C~13)~sN 0~1 P
0,15M (C~13) ~N HC03 Conversion of the tetramethylammonium hydroxide to TMA-HC03 was accomplished by bubbling a s~ream of C02 gas into the stirred solution until a pH of 8 was obtained. The pH o~ the solution can be changed by changing the amount of C02 added to the solution. The TMA-HC03 solution was diluted to the desired total carbon dioxide content (for example 30mM). This Tl~A-HC03 solution was then used to reconstitute previously lyophilized serum controls. Alternatively, ~he TMA bicarbonate or carbonate salt may be added to the lyophilized serum control prior to its reconstitution with distilled water.
These TMA-CH03-reconstituted controls were then analyzed by known analytical chemistry procedures. Many such control serums so reconstituted were analyzed in parallel with controls reconstituted with only distilled water. No significant differences between these two controls was found for any of the assays in which the controls were used, which included those mentioned above.
In another application oE our invention, we have found that other te-trasubstituted ammonium anion salts can be used in analytical chemical laboratories. For example, tetrasubsti-tuted ammonium halide salts can be used as standards and controls in both clinical and toxicology laboratories. Like the tetrasubs-tituted ammonium bicarbonate and carbonate salts, these saits add no color to reference materials to which they are added, _9_ :: . - -. . .. , . .: . . ~- . : . : ..... , : . - ~ . .~ . , ,:~ .: . :. - . ..... , : : . ~ , . . ; : . . : .
:~ : : . .: :,.. : , . .. . . . . .. .. . ....
so that they do not affect colorimetric determinations. 'l'hey are chemically unreactive in other laboratory methods, and they do not cause precipitation of any of the other components of the control materials to which they may be added, which could result in the plugging of instrumen ts.
Tetrabutyl ammonium bromide-reconstituted control serum is an application of our invention which may be used to fix the 10 level of bromide in bromide assays. A solution of 30 mM
tetrabutylammonium bromide was used to reconstitute lyophilized control serum, in a manner analogous to that used for the tetraalkylammonium bicarbonates. The resulting tetrabutylammonium bromide control material was the~ analyzed by known analytical chemistry procedures. These control serums were analyzed in parallel with controls reconstituted with distilled water only.
No significant differences between these two controls was found for any of the assays in which the controls were ~used, except for the halide assays, in which the tetrabutylammonium bromide-20 reconstituted controls were elevated by an amount correspondingto 30 mM bromide.
The tetrasubstituted ammonium anion salts of this invention are known in the art. However, these salts have not been used in the analytical chemistry procedures in which the salts of this invention are being used~ The salts and their methods of produckion are disclosed in Czechoslovakian Patent No. 157,000 and in the U.S. Patents Nos. 3,190,919, 3,579,581, 3,539j605 and .," ' ' :
_, :~
IO~
3,397,215. 'rhey arc also d:iscl.osed in the lite:rature in Mahajan and Rao, "Proton Maynetic Resonance and Internal Mokions in some Tetramethyl Ammonium Compounds", J. Phys. C: Solid State Phys., Vol. 7, Great sritain (1974) and the Ph.D Thesis of Teeter, Truman E. entitled "Reduction of Carbon Dioxide on Mercury Cathodes", the University of Oregon, Eugene, Oregon, June 1954 In order to show the efficacy of the quaternary ammonium carbonates and bicarbonates of this invention in adding carbon dioxide to a blood serum reference material, two samples from the same lot of lyophilized blood serum were taken. Each sample consisted of the amount of blood serum that could be lyophilized from 10ml of blood serum. 10ml of distilled water were added to the first sample, and the bottle containing the sample was agitated so that the serum would completely dissolve. The content of this sample was then analyzed with a conventional apparatus for analyzing blood serum. .:
: 20 The second sample of lyophilized blood serum was reconsti-tuted by adding 10ml of 22mM tetramethylammonium bicarbonate.
Here again, the mixture was slightly agitated so that the serum would be completely dissolved in the solution of tetramethylammon- : .
ium bicarbonate. The second sample was then analyzed using the same apparatus as used on the first sample.
The following chart gives the results of the tests for the constituents tested, and also shows the normal range for the constituents. -11-... .
~-~o~
VALUE PRIOR VAL~F. AFTER
TO OF
ADDITION OF ADDITION
TETR~METHYL- TETRAMEI'HYL-AMMONIUM AMMONIUM
CONSTITUENTSBICARBONATE BICARBONATE NORMAL RANGE
Sodium 150meq/1 150meq/1 138 - 146 Potassium 6.4meq/1 6.4meq/1 3.5 - 5.3 Chloride 107meq/1 107meq/1 97 - 108 Carbon Dioxide 3meq/1 25meq/124 - 32 Glucose 172mg/dl 171mg/dl 60 - 105 Urea Nitrogen 44mg/dl 44mg/dl 10 - 20 Creatinine 4.9mg/dl 4.9mg/dl 0;7 - 1.4 Calcium 11.9mg/dl 12.Omg/dl 8.5 - 10.5 Phosphorus 7.3mg/dl 7.3mg/dl 2.4 - 4.4 Total Protein 6.5mg/dl 6.4mg/dl 5.8 - 8.5 Alkaline Phosphatase 107IU/1 109IU/112 - 40 Bilirubin 3.6mg/dl 3.6mg/dl 0.2 - 1.2 Iron 198ug/dl 196ug/dl 70 - 200 Iron Binding Capacity 386ug/dl 412ug/dl 250 - 400 Cholesterol 153mg/dl 150mg/dl up to 260 (age dependent) Triglycerides 0.9meq/1 0.88meq/1 up to 2.15 (age dependent) Uric Acid 9.lmg/dl 9.0mg/dl 3 - 8 Aspartate Transaminase 63IU/1 61IU/16 - 38 Alanine Transaminase 165IU/1 166IU/19 - 25 ~: . , - : : . :: , ~ . :.;:
3S~L
V~LU~ Pl~IOR ~AWE AF'rER
rro OF
ADDI'I'[ON OF ADDI'I'ION
l'ETRAMF.'l~Ir TF'I'RAMEII~L-AMM~IUM AMMONIUM
CONSTITUENrSBICARBONATE BICMBONATE NORMPL RANGE
~actate Dehydrogenase 737IU/1 738IU/1 140 - 270 Creatine Kinase260IU/1 274IU/1 14 - 90 ~Iydroxybutyrate Dehydrogenase 587IU/1 591IU/1 110 - 230 Gamma Blutamyl rrranspeptidase26IU/1 26IU/1 4 - 40 Amylase 430U 434U 40 - 180 Notes: IU/l = international units per liter meq/l = milliequivalents/liter mg/dl = milligrams/deciliter ug/dl = micrograms/deciliter gm/dl = grams/deciliter ~0 It is thus seen from a review of the foregoing test results that, with the exception of the carbon dioxide, the addition of the -tetramethylammonium bicarbonate did not aect or interfere with the measurement of any oE the other constituents of the blood serum or contribute any of the o-ther constituents to the serum to change the concentration of the constituent in the serum. Insofar as the carbon dioxide is concerned, it can be seen rom the foregoing analysis that the concentration of carbon dioxide had fallen to 3meq/1 in the blood serum during lyophilization. This is from a normal range of 24 to 3am~q/1.
Ilowever, by mixing the lyophilized blood serum with the solution of tetramethyl-~mmonium bicarbonate, the concentration of carbon dioxide had risen to 25meq/1, which is within the normal range. Subsequent tests gave similar results.
:
The quaternary amines useul in this lnvention are all soluble in water. Additionally~ when used with blood serum, they ' ~lre al~o soluble in the serum. They are also soluble in the chemical mixtures used in the cllnical tests. Naturally, the ! amine should not interfere in any clinical assay. The co~ositions containing the amines should be s-terile. Sterility can be ob--tained by well known methods which will not affect the qua-ternary amines, for example, Eiltration of a solution of the amines through a filter which will retain particles the size of micro-10 organisms.
Among the cations which can be used in forming the amines of -this invention are the following:
~, trimethyl hydroxyethylammonium ~ dimethyl chloromethyl phenylammonium ,~ tetreathylammonium ,~ tetramethylammonium tetrapropylammonium tetrabutylammonium N, N - dimethyl - N - ethyl - p - nitrophenylammonium 20 N, N - dimethyl - N - fluoromethyl - N - phenylammonium N - cyclohexyl - N, N - dimethyl - N - isobutylammonium N - ethyl - N - methylpiperidinium N, N - dimethyl - N - ethyl - N - vinyl - ammonium N, N - diethyl - N - (2 - methoxyethyl) - N - n -butylammonium N - n - octyl - N, N, N - trimethylammonium N, N, N, N', N', N' - hexamethylethylene diaminium N - (2 - hydroxyethyl) trimethylammonium :.
The ~nions which cal~ be used are those speciied ~bove, which include carbonates, bicarbonates, a mixture of carbona-tes and bicarbonates, halides and any other anion of clincial interest.
In carrying o~t the method o this invention, -the lyophilized serum is reconsti-tu-ted with sufficien-t ammonium carbonate or bi-carbonate to provide from about 10 to about 40meq/1 of carbon dioxide in -the recons-tituted serum.
The reference serum oE the invention may be prepared by mixing a solution of the tetrasubstituted ammonium salt of the de-seired final concentra-tion with previously lyophillzed blood serum. Alternatively, if may be prepared by adding a predetermined amount of tetrasubstituted ammonium salt to the blood serum prior to lyophilization of the serum. To prepare the reference serum of of the invention by the latter method, the tetrasubstituted ammonium salt is added to pooled blood serum in sufficient amounts to provide the desited total CO2 level in the reconstituted serum.
For example, 1.0 ml of 0.4M tetrae-thyl ammonium bicarbonate is added to 10.0 ml of pooled blood serum. This mixture is then lyophilized by a known lyophilizatlon process. The lyophilization serum can then be reconstituted prior to use by dissolving it in 10.0 ml of distilled water, whereupon it can be analyzed by known procedures.
Without further elaboration, the foregoing will so fully illustrate our invention, that others may, by applying current or Euture knowledge, readily adapt the same for use under various conditions of service.
: 20 The second sample of lyophilized blood serum was reconsti-tuted by adding 10ml of 22mM tetramethylammonium bicarbonate.
Here again, the mixture was slightly agitated so that the serum would be completely dissolved in the solution of tetramethylammon- : .
ium bicarbonate. The second sample was then analyzed using the same apparatus as used on the first sample.
The following chart gives the results of the tests for the constituents tested, and also shows the normal range for the constituents. -11-... .
~-~o~
VALUE PRIOR VAL~F. AFTER
TO OF
ADDITION OF ADDITION
TETR~METHYL- TETRAMEI'HYL-AMMONIUM AMMONIUM
CONSTITUENTSBICARBONATE BICARBONATE NORMAL RANGE
Sodium 150meq/1 150meq/1 138 - 146 Potassium 6.4meq/1 6.4meq/1 3.5 - 5.3 Chloride 107meq/1 107meq/1 97 - 108 Carbon Dioxide 3meq/1 25meq/124 - 32 Glucose 172mg/dl 171mg/dl 60 - 105 Urea Nitrogen 44mg/dl 44mg/dl 10 - 20 Creatinine 4.9mg/dl 4.9mg/dl 0;7 - 1.4 Calcium 11.9mg/dl 12.Omg/dl 8.5 - 10.5 Phosphorus 7.3mg/dl 7.3mg/dl 2.4 - 4.4 Total Protein 6.5mg/dl 6.4mg/dl 5.8 - 8.5 Alkaline Phosphatase 107IU/1 109IU/112 - 40 Bilirubin 3.6mg/dl 3.6mg/dl 0.2 - 1.2 Iron 198ug/dl 196ug/dl 70 - 200 Iron Binding Capacity 386ug/dl 412ug/dl 250 - 400 Cholesterol 153mg/dl 150mg/dl up to 260 (age dependent) Triglycerides 0.9meq/1 0.88meq/1 up to 2.15 (age dependent) Uric Acid 9.lmg/dl 9.0mg/dl 3 - 8 Aspartate Transaminase 63IU/1 61IU/16 - 38 Alanine Transaminase 165IU/1 166IU/19 - 25 ~: . , - : : . :: , ~ . :.;:
3S~L
V~LU~ Pl~IOR ~AWE AF'rER
rro OF
ADDI'I'[ON OF ADDI'I'ION
l'ETRAMF.'l~Ir TF'I'RAMEII~L-AMM~IUM AMMONIUM
CONSTITUENrSBICARBONATE BICMBONATE NORMPL RANGE
~actate Dehydrogenase 737IU/1 738IU/1 140 - 270 Creatine Kinase260IU/1 274IU/1 14 - 90 ~Iydroxybutyrate Dehydrogenase 587IU/1 591IU/1 110 - 230 Gamma Blutamyl rrranspeptidase26IU/1 26IU/1 4 - 40 Amylase 430U 434U 40 - 180 Notes: IU/l = international units per liter meq/l = milliequivalents/liter mg/dl = milligrams/deciliter ug/dl = micrograms/deciliter gm/dl = grams/deciliter ~0 It is thus seen from a review of the foregoing test results that, with the exception of the carbon dioxide, the addition of the -tetramethylammonium bicarbonate did not aect or interfere with the measurement of any oE the other constituents of the blood serum or contribute any of the o-ther constituents to the serum to change the concentration of the constituent in the serum. Insofar as the carbon dioxide is concerned, it can be seen rom the foregoing analysis that the concentration of carbon dioxide had fallen to 3meq/1 in the blood serum during lyophilization. This is from a normal range of 24 to 3am~q/1.
Ilowever, by mixing the lyophilized blood serum with the solution of tetramethyl-~mmonium bicarbonate, the concentration of carbon dioxide had risen to 25meq/1, which is within the normal range. Subsequent tests gave similar results.
:
The quaternary amines useul in this lnvention are all soluble in water. Additionally~ when used with blood serum, they ' ~lre al~o soluble in the serum. They are also soluble in the chemical mixtures used in the cllnical tests. Naturally, the ! amine should not interfere in any clinical assay. The co~ositions containing the amines should be s-terile. Sterility can be ob--tained by well known methods which will not affect the qua-ternary amines, for example, Eiltration of a solution of the amines through a filter which will retain particles the size of micro-10 organisms.
Among the cations which can be used in forming the amines of -this invention are the following:
~, trimethyl hydroxyethylammonium ~ dimethyl chloromethyl phenylammonium ,~ tetreathylammonium ,~ tetramethylammonium tetrapropylammonium tetrabutylammonium N, N - dimethyl - N - ethyl - p - nitrophenylammonium 20 N, N - dimethyl - N - fluoromethyl - N - phenylammonium N - cyclohexyl - N, N - dimethyl - N - isobutylammonium N - ethyl - N - methylpiperidinium N, N - dimethyl - N - ethyl - N - vinyl - ammonium N, N - diethyl - N - (2 - methoxyethyl) - N - n -butylammonium N - n - octyl - N, N, N - trimethylammonium N, N, N, N', N', N' - hexamethylethylene diaminium N - (2 - hydroxyethyl) trimethylammonium :.
The ~nions which cal~ be used are those speciied ~bove, which include carbonates, bicarbonates, a mixture of carbona-tes and bicarbonates, halides and any other anion of clincial interest.
In carrying o~t the method o this invention, -the lyophilized serum is reconsti-tu-ted with sufficien-t ammonium carbonate or bi-carbonate to provide from about 10 to about 40meq/1 of carbon dioxide in -the recons-tituted serum.
The reference serum oE the invention may be prepared by mixing a solution of the tetrasubstituted ammonium salt of the de-seired final concentra-tion with previously lyophillzed blood serum. Alternatively, if may be prepared by adding a predetermined amount of tetrasubstituted ammonium salt to the blood serum prior to lyophilization of the serum. To prepare the reference serum of of the invention by the latter method, the tetrasubstituted ammonium salt is added to pooled blood serum in sufficient amounts to provide the desited total CO2 level in the reconstituted serum.
For example, 1.0 ml of 0.4M tetrae-thyl ammonium bicarbonate is added to 10.0 ml of pooled blood serum. This mixture is then lyophilized by a known lyophilizatlon process. The lyophilization serum can then be reconstituted prior to use by dissolving it in 10.0 ml of distilled water, whereupon it can be analyzed by known procedures.
Without further elaboration, the foregoing will so fully illustrate our invention, that others may, by applying current or Euture knowledge, readily adapt the same for use under various conditions of service.
Claims (21)
EXCLUSIVE PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED
AS FOLLOWS:
1. A composition capable of use as a component of a clinical control material, comprising water and at least one water-soluble quaternary ammonium carbonate or bi-carbonate, said composition including no substance which will affect assays for sodium, potassium, calcium, ammo-nium, chloride and iron ions.
2. The composition of claim 1 wherein the quaternary ammonium carbonate or bicarbonate is in concentration sufficient to provide CO2 concentration of from about 10 to about 40 meq./l.
3. The composition of claim 1 wherein the amount of sodium, potassium, calcium, ammonium, chloride and iron ions is, respectively, less than about 2 millimolar, about 0.25 millimolar, about 0.5 mg/100 ml, 0.5 µg./100 ml, 3 millimolar and 3 µg./100 ml.
4. The composition of claim 3 wherein the quaternary ammonium carbonate or bicarbonate is in concentration sufficient to provide a CO2 concentration of from about 10 to about 40 meq./l.
5. The composition of claim 1 wherein the amount of sodium, potassium, calcium, ammonium, chloride and iron ions is, respectively, less than about 1 millimolar, 0.1 millimolar, about 0.2 mg/100 ml, 0.2 µg/100 ml, 2 millimolar and 2 µg/100 ml.
6. The composition of claim 5 wherein the quaternary ammonium carbonate or bicarbonate is in concentration sufficent to provide a CO2 concentration of from about 10 to about 40 meq./l.
7. A sterile composition capable of use as a component of a clinical control material, comprising water and at least one water-soluble quaternary ammonium carbonate or bicarbonate.
8. The composition of claim 7 wherein the quaternary ammonium carbonate or bicarbonate is in concentration sufficient to provide a CO2 concentration of from about 10 to 40 meq./l.
9. A composition capable of use as a component of a clinical control material, comprising water and at least one water-soluble quaternary ammonium carbonate or bi-carbonate, said composition including no substance which will affect assays for sodium, potassium, chloride, iron, phosphorus and calcium ions, urea, uric acid, creatinine, glucose, cholesterol, bilirubin, total protein, lactic acid dehydrogenase, amylase, acid phosphatase, alkaline phosphatase, thyroid hormone uptake and total thyroid hormone.
10. The composition of claim 9 wherein the quaternary ammonium carbonate of bicarbonate is in concentration sufficent to provide a CO2 concentration of from about 10 to about 40 meq./l.
11. A composition capable of use as a clinical control material, consisting essentially of at least one water-soluble quaternary ammonium carbonate or bicarbonate and at least one animal body fluid.
12. A dry composition comprising at least one dried animal body fluid and at least one water-soluble quater-nary ammonium carbonate or bicarbonate.
13. The composition of claim 12 wherein the amount of salt is known.
14. The composition of claim 12 wherein the body fluid is serum.
15. The composition of claim 12 wherein the composition is in vitro.
16. A composition for use as a clinical control serum, comprising at least one animal body fluid and at least one water-soluble quaternary ammonium carbonate or bi-carbonate, wherein each tetrasubstituted group of said quaternary ammonium carbonate or bicarbonate is selected from the group consisting of saturated heterocycles, un-saturated heterocycles, cycloalkenyl groups, cycloalkyl groups, and branch chain lower alkyls, normal lower alkyls and normal lower alkenyls, with each of said lower alkyls and alkenyls having eight or less carbon atoms.
17. A method of making a clinical control serum which comprises mixing a water-soluble quaternary ammonium carbonate or bicarbonate and an animal body fluid, where-in each tetrasubstituted group of said quaternary ammon-ium carbonate or bicarbonate is selected from the group consisting of saturated heterocycles, unsaturated hetero-cycles, cycloalkenyl groups, cycloalkyl groups, and branch chain lower alkyls, normal lower alkyls and normal lower alkenyls, with each of said lower alkyls and alkenyls having eight or less carbon atoms.
18. A method for making a clinical control material which comprises mixing a quaternary ammonium carbonate or bicarbonate and a dry animal body fluid.
19. A method for making a clinical control material which comprises mixing an aqueous solution of a water-soluble quaternary ammonium carbonate or bicarbonate and an animal body fluid and then lyophilizing the mixture.
20. The method of claim 19 wherein the lyophilized mix-ture is reconstituted in water and then assayed for at least one of its clinically significant components.
21. The method of claim 18 wherein the quaternary ammo-nium carbonate or bicarbonate is added in sufficient concentration to provide from about 10 to about 40 meq./l.
of CO2 in the mixture when the dry animal body fluid is reconstituted to its water content prior to being dried.
of CO2 in the mixture when the dry animal body fluid is reconstituted to its water content prior to being dried.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US68924276A | 1976-05-24 | 1976-05-24 | |
| US689,242 | 1976-05-24 | ||
| US794,864 | 1977-05-09 | ||
| US05/794,864 US4141856A (en) | 1976-05-24 | 1977-05-09 | Reference material for establishing anion concentrations in analytical chemistry tests |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1088851A true CA1088851A (en) | 1980-11-04 |
Family
ID=27104369
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA278,995A Expired CA1088851A (en) | 1976-05-24 | 1977-05-24 | Reference material for establishing anion concentrations in analytical chemistry tests |
Country Status (10)
| Country | Link |
|---|---|
| JP (1) | JPS534598A (en) |
| AU (1) | AU517248B2 (en) |
| CA (1) | CA1088851A (en) |
| DE (1) | DE2723070C2 (en) |
| ES (1) | ES459675A1 (en) |
| FR (1) | FR2353060A1 (en) |
| GB (1) | GB1586582A (en) |
| IT (1) | IT1084035B (en) |
| NL (1) | NL7705671A (en) |
| SE (1) | SE446129B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4264471A (en) * | 1979-06-04 | 1981-04-28 | E. I. Du Pont De Nemours And Company | Serum and plasma clarification process |
| CN103917643B (en) * | 2011-11-22 | 2020-06-05 | 西门子医疗保健诊断公司 | Apparatus comprising dry reagents for reconstitution into calibration and/or quality control solutions, and methods of making and using same |
| JP7110360B2 (en) | 2017-10-09 | 2022-08-01 | テルモ ビーシーティー バイオテクノロジーズ，エルエルシー | Freeze-drying method |
| JP7495426B2 (en) | 2019-03-14 | 2024-06-04 | テルモ ビーシーティー バイオテクノロジーズ，エルエルシー | Filling tool for freeze-drying containers, system and method of use |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3466249A (en) * | 1967-02-13 | 1969-09-09 | Baxter Laboratories Inc | Blood serum reference standard for multi-automated analytical procedures |
| US3629142A (en) * | 1969-12-08 | 1971-12-21 | Edward P Marbach | Reference standard blood serum for the calibration of automatic blood serum analyzing apparatus |
| DE2324386C2 (en) * | 1973-05-14 | 1975-07-03 | Boehringer Mannheim Gmbh, 6800 Mannheim | Aqueous cholestrin standard |
| US3861255A (en) * | 1973-11-23 | 1975-01-21 | Cray L Ross | Device for cutting cavities |
-
1977
- 1977-05-21 DE DE19772723070 patent/DE2723070C2/en not_active Expired
- 1977-05-23 FR FR7715686A patent/FR2353060A1/en active Granted
- 1977-05-23 SE SE7705987A patent/SE446129B/en not_active IP Right Cessation
- 1977-05-24 JP JP6034277A patent/JPS534598A/en active Granted
- 1977-05-24 GB GB2195177A patent/GB1586582A/en not_active Expired
- 1977-05-24 CA CA278,995A patent/CA1088851A/en not_active Expired
- 1977-05-24 AU AU25449/77A patent/AU517248B2/en not_active Expired
- 1977-05-24 IT IT2391277A patent/IT1084035B/en active
- 1977-05-24 NL NL7705671A patent/NL7705671A/en not_active Application Discontinuation
- 1977-06-02 ES ES459675A patent/ES459675A1/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| GB1586582A (en) | 1981-03-18 |
| ES459675A1 (en) | 1978-12-01 |
| JPS6314303B2 (en) | 1988-03-30 |
| JPS534598A (en) | 1978-01-17 |
| FR2353060A1 (en) | 1977-12-23 |
| SE7705987L (en) | 1977-11-25 |
| IT1084035B (en) | 1985-05-25 |
| AU2544977A (en) | 1978-11-30 |
| FR2353060B1 (en) | 1983-08-05 |
| AU517248B2 (en) | 1981-07-16 |
| DE2723070A1 (en) | 1977-12-15 |
| DE2723070C2 (en) | 1986-03-06 |
| NL7705671A (en) | 1977-11-28 |
| SE446129B (en) | 1986-08-11 |
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