IL46338A - Determination of serum creatinne - Google Patents

Determination of serum creatinne

Info

Publication number
IL46338A
IL46338A IL46338A IL4633874A IL46338A IL 46338 A IL46338 A IL 46338A IL 46338 A IL46338 A IL 46338A IL 4633874 A IL4633874 A IL 4633874A IL 46338 A IL46338 A IL 46338A
Authority
IL
Israel
Prior art keywords
sodium
sulfate
creatinine
reagent
detergent
Prior art date
Application number
IL46338A
Other versions
IL46338A0 (en
Original Assignee
Coulter Diagnostics Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Coulter Diagnostics Inc filed Critical Coulter Diagnostics Inc
Publication of IL46338A0 publication Critical patent/IL46338A0/en
Publication of IL46338A publication Critical patent/IL46338A/en

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/70Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving creatine or creatinine
    • YGENERAL 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
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00Chemistry: analytical and immunological testing
    • Y10T436/10Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
    • YGENERAL 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
    • Y10TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T436/00Chemistry: analytical and immunological testing
    • Y10T436/14Heterocyclic carbon compound [i.e., O, S, N, Se, Te, as only ring hetero atom]
    • Y10T436/145555Hetero-N
    • Y10T436/147777Plural nitrogen in the same ring [e.g., barbituates, creatinine, etc.]

Landscapes

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

Abstract

A colorimetric process for the quantitative determination of creatinine in blood serum or urine wherein an alkaline pH in the approximate range of 11.0 to 13.5 is maintained during colorimetry evaluation and in which urea and a detergent are caused to react simultaneously and synergistically with the serum or urine protein to prevent extraneous chromogen formation in the conventional alkaline-picrate reaction for creatinine. [US3894843A]

Description

DETERMINATION OP SERUM CREATININE DETERMINATION OF SERUM CREATININE A colorimetric process for the quantitative determination of creatinine in blood serum or urine wherein an alkaline pH in the approximate range of 11.0 to 13.5 is maintained during colorimetry evaluation and in which urea and a · detergent is caused to react simultaneously and synergistically with the serum or urine protein to prevent extraneous chromogen formation in the conventional alkaline-picrate reaction for creatinine.
BACKGROUND OF THE INVENTION This invention relates to the analysis of body fluids for the quantitative determination of the creatinine conteni^ of the fluid, such as blood serum and urine.
. The quantitative determination of creatinine is a very valuable procedure in connection with biologic fluids such as blood serum' and urine, for the diagnosis of certain diseases. Creatinine is a waste product removed from the blood stream by the kidneys. Since creatinine is excreted only by way of the kidneys, for instance, kidney disease can be diagnosed from an increased creatinine content measured in the serum.
Quantitative creatinine determination also is useful in the diagnosis of urinary obstruction, intestinal obstruction,, and nephritis,, an inflamation of the kidneys caused by infection, the degenerative process or vascular disease.. In U.S. patent No. 3,705,013, there is discussed some features of critical creatinine determinations desired for accurate and effective diagnosis of diseases.
U.S. Patents 3,557,018 and 3,705,013 discuss many known methods for determination of creatinine in biologic fluids.
DESCRIPTION OF THE PRIOR ART Most current methods for the determination of creatinine in serum or urine rely on the development of a red-orange ^ color formed by the interaction of alkaline-picrate and creatinine, first reported by M. Jaffe in 1886 and applied by. Otto Folin to determination of urinary creatinine in 1904. Since that time, this method using the so-called "Jaffe reaction" has become the most commonly used method for creatinine analysis.
The Jaffe reaction is not considered specific for .. creatinine in the presence of protein, glucose, and a number of unknown substances present in normal human sera. Methods designed to improve specificity have, for the most part, relied on use of protein-free filtrates. In addition to utilization of protein-free filtrates, so-called "specific methods" for creatinine have been introduced in which the filtrate is used in adsorption-elution. techniques; see Brod, J. and Kotatko, J., J. Cas. Lek. Ces. 88,665 - Excerpta Medica, Amst. 3,477 (1950); Hare, R.S., Proc. Soc. Exp. Biol. N.Y., 74,148 (1950); Haugen, H.N. and Blegen, E.M., Scand.
J. Clin. Invest. 5,58 (1953) ; and Polar, E. and Metkoff, J., Clin. Chem. 11,763 (1965). The most common absorption media used have been hydrated aluminum silicate and ion-exchange ■ resin.
.The problems encountered with adsorption-elution techniques center around complex procedures to effect the adsorption and eliminated. - Alternate "specific" techniques have utilized reagents such as eerie sulfate with protein-free filtrates to obtain "destruction of interfering substances". See Kostir, J.V. and Rabek, V., Biochim. Biophys. Acta 5, 210 (1950) Kostir, J.V. and Sonka, J., Biochim. Biophys. Acta 8,86 (1952) and use of "creatinine destroying bacteria" - Dubos, R. and Miller, V.F., J. Biol. Chem. 121,427 (1937). The latter procedure is one in which serum protein-free filtrates are divided into two aliquots; one aliquot being treated with the "creatinine destroying bacteria", the other aliquot left untreated. Sub¬ sequent Jaffe reactions with both aliquots give different d al^sorbance readings. Subtracting the "bacteria-treated" from the "non-treated" aliquot value is assumed to yield a "true creatinine" value. Specificity cannot be satisfactorily proven by this method either.
More recently, attempts have been made to develop Jaffe reaction methods avoiding serum deproteinization which fall into two primary categories. The first involves a so-called "kinetic procedure" requiring critically-timed measurement of rate of formation of the reddish-orange Jaffe color represented by Larsen, K. , Clin. Chim. Acta 41, 209 (1972 ) . Problems arise with the need for relatively sophisticated kinetic measurement spectrophotometers and use of extremely short color development times (30-120 seconds) . Effects of interference by non-creat- inine, Jaffe-reactive chromogens cannot be entirely disproven, as appears from Raabo, E. and Walloe-Hansen, P., Scand. J. Clin 137,829 Lab. Invest. 29,297 (1972) and Heinegard, D. and Tiderstrorn, "" G. , Clin, Chim.. Acta 43,305 (1973).
The second avenue of approach consists of introduction^ of suppression of non-creatinine, Jaffe-positive chromogens 5 by varying reaction pH or incorporating reagents shown to suppress the effect of color interference by non-creatinine substances. Reagent blanks or running serums at two different pH's are an integral part of all of these approaches as • seen from Raabo, E. and Walloe-Hansen, P. Scand. J. Clin. Lab. 10 Invest. 29,297 (1972) ; Heinegard, D. and Tiderstrorn, G., Clin. Chim. Acta 43,305 (1973). The methods relying solely on the assumption that running parallel reactions on the same serum, one at a higher pH than the other, subtracting the low pH value from that of the higher pH, and then using 15 "the factor 2.3" as a correction for protein interference, as taught by Raabo, E. and Walloe-Hansen, P., Scand. J. Clin.
Lab. Invest. 29,297 (1972), based on a large number of comparisons with a previous specific method as taught by Brod, J. and Sirota, J. H., J. Clin. Invest. 27,645 (1948) leaves doubts .20 about accuracy primarily because of this statistically observed "2.3 factor" and complete reliance on specificity based on the two separate pH dependent reactions.
SUMMARY OF THE INVENTION With the above delineated problems of the prior art in mind, the invention provides a procedure for the detecti¾ creatinine which employs an alkaline picrate reagent which substitutes sodium phosphate for the more commonly used sodium hydroxide or lithium hydroxide. Urea is alkane OK incorporated into the alkaline picrate reagent. An aralkane sulfate detergent of sodium, potassium or lithium, such as sodium dodecyl sulfate (sodium lauryl sulfate) and sodium borate are added to the reagent. In this procedure, suppressio of interfering chromogens is maximized and the serum blank is eliminated.
It is believed that in the presence, of urea, the protein molecules change in configuration and unfold as a result of mild denaturation. This configuration change increases exposure of the protein molecule to binding with the sodium dodecyl sulfate with resulting decrease in blank activity.
Test results using the invention have correlated well with reference methods using filtrates and adsorption techniques. 9 DESCRIPTION OF SPECIFIC EMBODIMENTS The following description is given to enable persons skilled in the art to more clearly understand and practice the invention. They should not be considered as limitations . upon the scope of the invention but should be regarded as illustrative only.
The reagent formulations are as follows: A. Saturated picric acid solution - approximatel 15 grams added to warm distilled water until no , further dissolving is observed.
B. i. Sodium dodecyl sulfate solution - 4 grams/100 ml. ii. Sodium borate in sodium or potassium phosphate - ~~ ···.,; ... dibasic solution - 0.05 M of borate to, 0.05 M ·■·.'"■■ · phosphate. ■·,■'■ iii. Urea solution - 40 grams/100 ml.
In using these stock or prepared solutions, a first working solution is prepared consisting of two parts of the 40% urea solution combined with two parts of the 4% sodium dodecyl sulfate solution. A preferred alternate working . solution consists of one part of the urea solution with three parts of the sodium dodecyl sulfate solution. Then, one part of a said sodium dodecyl sulfate - urea solution is mixed with one part of the borate-phosphate buffer solution and buffered using sodium hydroxide to a pH of about 12.4. The second working solution comprises the saturated picric acid solution.
Prior to use, four parts of the first working solution are mixed with one part of the saturated picric acid solution to provide a single alkaline picrate reagent.
In the creatinine determination procedure using the invention, 0.01 to 2.0 ml of sample has been used with 0.2 to 25.0 ml of final mixed reagent. Preferably, however, 0.1 to 0.2 ml of sample with 4.0 ml of final reagent is used.
In this preferred procedure, 4.0 ml of alkaline picrate reagen "is introduced into a suitable test tube and pre-incubated at- 37° Centrigrade for 3 to 5 minutes. Then, 0.1 or 0.2 ml of sample is mixed into the reagent and incubation at 37° Centrigrade maintained for from 10 to 30 minutes. Preferably, such incubation is maintained for 15 minutes.
^ The incubated solution then is introduced into a colorime or spectrophotometer against a blank and light absorpt of the thusly formed creatinine picrate is read at 490 to 535 nm. Preferably, the reading is made at 500 nm. Then, one ca calculate the value of unknown against primary aqueous standard or secondary protein-based standard run in conjunctio with the unknown using the following formula: (1) Value of Unknown ?η¾η^"^, * value of standar O.D. Standard The specific example given above is capable of being varied in certain respects within the parameters of the invention. The alkaline picrate reagent can be buffered to within a pH range of 11.0 to 13.5 by selective mixing of the buffer solution, to wit, the sodium borate in the sulfate solution as mentioned. Optical density measurements can be other modifications may occur to the skilled artisan relating to a particular material formulation to utilize the essential teachings of the invention. For instance, other detergents which may be used are: sodium octyl sulfate, sodium tetradecyl sulfate, sodium heptadecyl sulfate, sodium cetyl sulfate, sodium myristyl sulfate, sodium octadecyl sulfate, sodium oleyl sulfate, sodium tridecyl sulfate, potassium lauryl sulfate and dodecyl benzene sulfate.
The analagous potassium or lithium salts are equally useful.. Generally, the detergent may be characterized as an aralkane sulfate of sodium, potassium or lithium.
What is claimed is:

Claims (15)

Appin*No, 46338/2 CLalMSt Ψ
1. » A colometric process for the determination of creatinine in a biologic fluid sample containing proteins comprising, A. adding an alkaline piorate reagent to the biologic fluid which forms a colored complex having a characteristic wavelength of maximum light absorption with the creatinine in said fluid, adding ftirther an alkane or aralkane sulfate detergent of sodium potassium or lithium, while preventing the interference of protein wit the desired determination by forming urea-protein complexes which prevent formation of extraneous chroinogeus of a non-croatiuine character) B« measuring the degree of light absorption at said characteristic wavelength; and C. determining the creatinine concentration in said fluid by comparing the measured degree Of light absorption with values of light absorption obtained from standard aqueous solutions containing the reagent and known creatinine concentrations*
2. The process of claim 1 vherein ure is added to the sample prior to oolorimetric determinations being made in an amount sufficient to form such protein complexes* (
3. The process of claim 2 wherein a detergent is added to the urea and the combination is then added to sample
4. The process of claim 3 wherein the detergent selected is sodium octyl sulfate.
5. The process of claim 3 wherein the detergent selected is sodium tetradecyl sulfate.
6. The process of claim 3 wherein the detergent selected is sodium heptadecyl sulfate. (
7. The process of claim 3 wherein the detergent selected is sodium cetyl sulfate.
8. The process of claim 3 wherein the detergent selected is sodium dodecyl sulfate.
9. The process of claim 3 wherein the detergent selected is sodium myristyl sulfate.
10. The process of claim 3 wherein the detergent selected is. sodium octadecyl sulfate. ( 137,829
11. The process of claim 3 wherein the detergent selected is sodium oleyl sulfate.
12. The process of claim 3 wherein the detergent selected is sodium tridecyl sulfate.
13. The process of claim 3 wherein the detergent selected is dodecyl benzene sulfate.
14. The process of claim 3 wherein the detergent alkane or comprises a sodium, potassium or lithium/aralkane sulfate solution. )
15. The process of claim 3 wherein sample size of sample is maintained from 0.01 to 2.0 ml. The process of claim 3 wherein the volume of reagent is maintained from 0.2 to 25.0 ml. The process of claim 3 wherein pH is maintained from 11.0 to 13.5 during colorimetry. The process of claim 3 wherein a buffer is added to the reagent to maintain a pH of between 11.0 to 13.5 during colorimetry. ( A reagent system for use in the determination of creatinine in a biological fluid sample containing proteins and in which the creatinine concentration in the sample is determined by comparing the measured degree of light absorption with values of light absorption obtained from standard solutions/ comprising: A. Saturated picric acid solution in distilled water; and B. i. urea solution; ii. a buffered solution of sodium borate in a sulphate salt of sodium or potassium buffered to a pH of between 11.0 to 13.5; and alkane or iii. an/aralkane sulphate detergent solution. The reagent system of claim 19 in which. the solution of Part A is mixed with the solution of Part B to provide an alkaline picrate reagent used in the creatinine determination. ( ( 137,829 The reagent system as claimed in claim 20 in which the alkaline picrate reagent is pre- incubated at 37° C. and then the sample is introduced into the reagent and incubated at for from 10 to 30 inubes. Tel-Aviv, December 24, 1974
IL46338A 1974-03-04 1974-12-25 Determination of serum creatinne IL46338A (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US447864A US3894843A (en) 1974-03-04 1974-03-04 Determination of serum creatinine

Publications (2)

Publication Number Publication Date
IL46338A0 IL46338A0 (en) 1975-03-13
IL46338A true IL46338A (en) 1977-10-31

Family

ID=23778053

Family Applications (1)

Application Number Title Priority Date Filing Date
IL46338A IL46338A (en) 1974-03-04 1974-12-25 Determination of serum creatinne

Country Status (12)

Country Link
US (1) US3894843A (en)
JP (1) JPS50120897A (en)
BE (1) BE824205A (en)
CH (1) CH594245A5 (en)
DE (1) DE2504994A1 (en)
ES (1) ES434127A1 (en)
FR (1) FR2263515A1 (en)
GB (1) GB1445954A (en)
IL (1) IL46338A (en)
NL (1) NL7416476A (en)
SE (1) SE7501016L (en)
ZA (1) ZA75224B (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4111657A (en) * 1977-01-21 1978-09-05 American Monitor Corporation Creatinine assay and reagent system
US4529708A (en) * 1983-04-07 1985-07-16 American Monitor Corporation Assay for the determination of creatinine
US4539295A (en) * 1983-06-30 1985-09-03 Beckman Instruments, Inc. Binary kinetic assay method and apparatus
US4818703A (en) * 1985-10-23 1989-04-04 Pizzolante John M Stabilized alkaline picrate reagent for jaffe creatinine determination
US4950611A (en) * 1987-06-26 1990-08-21 Beckman Instruments Cold stable liquid creatinine reagent
CN101466875B (en) * 2006-06-12 2011-01-05 日矿金属株式会社 Rolled copper or copper alloy foil having roughened surface and roughening method of the rolled copper or copper alloy foil
SG10201913421YA (en) 2014-02-28 2020-03-30 Nitto Denko Corp Urinalysis device and dry reagent for quantitative urinalysis

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1648977A1 (en) * 1967-07-20 1970-08-13 Merck Anlagen Gmbh Color reagent for the determination of creatinine
US3705013A (en) * 1970-01-05 1972-12-05 Xerox Corp Analytical procedures and compositions therefor
US3682586A (en) * 1971-03-10 1972-08-08 Union Carbide Corp Process for the determination of creatinine body fluids

Also Published As

Publication number Publication date
IL46338A0 (en) 1975-03-13
GB1445954A (en) 1976-08-11
ES434127A1 (en) 1977-03-01
DE2504994A1 (en) 1975-09-11
AU7596374A (en) 1976-06-03
SE7501016L (en) 1975-09-05
CH594245A5 (en) 1977-12-30
ZA75224B (en) 1976-08-25
FR2263515A1 (en) 1975-10-03
BE824205A (en) 1975-07-08
US3894843A (en) 1975-07-15
NL7416476A (en) 1975-09-08
JPS50120897A (en) 1975-09-22

Similar Documents

Publication Publication Date Title
Trivedi et al. New enzymatic method for serum uric acid at 500 nm.
Schade et al. Bound iron and unsaturated iron-binding capacity of serum; rapid and reliable quantitative determination
Iwata et al. New micro-turbidimetric method for determination of protein in cerebrospinal fluid and urine.
US3950226A (en) Novel reagent and method for the determination of urea in biological fluids
Rej et al. Interference by Tris buffer in the estimation of protein by the Lowry procedure
Walser Determination of free magnesium ions in body fluids. Improved methods for free calcium ions, total calcium, and total magnesium
EP0215170B1 (en) Single color reading method for determining fructosamine
WO1992011524A2 (en) Reagent and methods for calcium determination
US3894843A (en) Determination of serum creatinine
JPH0425000B2 (en)
CA1089338A (en) Creatinine assay and reagent system
Doumas et al. Chemical nature of a synthetic bilirubin conjugate and its reactivities in the total and direct reactions by the Jendrassik-Gróf method.
JPWO2002086151A1 (en) Biological component measurement method
Kekki Microdetermination of Amino Nitrogen as Copper Complexes a Modification for Plasma and Urine
US4529708A (en) Assay for the determination of creatinine
US5219760A (en) Process for the determination of iron
Van Pilsum Determination of creatinine and related guanidinium compounds
US4211531A (en) Colorimetric cholesterol assay
Lawrie The excretion of l (—)-tyrosine in urine
US3778384A (en) Diagnostic composition for the quantitative determination of glucose
JPH0690207B2 (en) Method and combined reagent for quantifying iron in serum
Ujiie et al. A new micromethod for colorimetric determination of zinc in serum
Harrison et al. Biomass measurements: protein
Iron Panel of the International Committee for Standardization in Haematology Revised recommendations for the measurements of the serum iron in human blood
JP2589088B2 (en) Determination of anti-streptolysin O