JPH05196616A - Reagent composition for measuring ionic strength or specific gravity of aqueous liquid specimen - Google Patents

Abstract

PURPOSE:To facilitate visual judgment and enable easily automatic measurement of a number of liquids by containing at least one of pH buffers, at least one of pH indicators, and at least one of surfactants as asensitizing agent. CONSTITUTION:The reagent composition contains at least one of pH buffers, at least one of pH indicators, and at least one of surfactants as a sensitizing agent. As the pH buffers, a material used in a predetermined range, such as sodium chrolide, is used. The pH indicators are indicators whose transition interval lies within the pH variation width of the pH buffers, and are preferably triphenylenemethanic pH indicators, in which the dissociation is activated by the reinforcement of the ionic strength, for example bran thymol blue. As the surfactants, a material containing such function that the dissociation of the pH indicators activated by the reinforcement of the ionic strength is suppressed, for example an anionic active agent, is preferred. The agent composition and an aqueous liquid specimen are mixed together, and the variation in the pH of the solution and the variation in the degree of dissociation of the pH indicators are detected so that ionic strength or specific gravity of the aqueous liquid specimen is measured.

Description

Detailed Description of the Invention

TECHNICAL FIELD The present invention relates to a reagent composition for measuring the ionic strength or specific gravity of an aqueous liquid sample, and a test piece impregnated, coated or applied with the reagent composition.

[0002]

BACKGROUND OF THE INVENTION As a method currently generally used as a method for directly measuring the specific gravity of urine, a liquid hydrometer, a urine hydrometer,
There is a method of using a pycnometer and a refractometer. Most of these methods have achieved the required accuracy, but these devices require time calibration, instrument cleaning control to maintain accuracy, and time and It is expensive and has many inconveniences. Further, at the time of measurement, the sample amount of urine needs to be a certain amount or more, and it may be insufficient. Further, it is difficult to read the scale due to bubbles on the liquid surface or capillary action.

On the other hand, a method of measuring changes in color tone and absorbance caused by mixing urine and a test solution with the naked eye or a colorimeter, or by dipping a test piece processed into a stick into urine, Recently, a method for visually determining a change in color tone and simply measuring the specific gravity and ionic strength of urine has been developed and is partially put to practical use. This method can be carried out by measuring the reflectance by a light emitting diode, as well as by measuring the absorbance with a colorimeter and the naked eye determination by comparison with a color chart. Further, the method using the test piece is combined with other urine test items, for example, glucose, hemoglobin, protein, leukocyte, bilirubin, pH, ascorbic acid, urobilinogen, ketone body, nitrite, etc. It has the advantage that it can be measured simultaneously.

The methods disclosed so far as the methods for measuring the ionic strength or the specific gravity of a liquid sample for measuring the change in color tone and the change in absorbance are listed below.
In U.S. Pat.No. 4,015,462, a carrier matrix contains microcapsules containing a colorant, and when the microcapsules contact a low osmotic pressure solution, the pressure inside the microcapsules increases, the microcapsules expand and burst, and Describes a method (the color strength is proportional to the specific gravity of the solution) that utilizes the phenomenon that is dissolved and the matrix is discolored.

JP-A-55-101047 discloses at least about 50 such as polyacrylic acid or polyvinylamine.
There is described a method for measuring ionic strength or specific gravity using a test piece composed of a composition containing a polyelectrolyte which is% neutralized and a pH indicator.

Japanese Patent Laid-Open No. 56-21064 discloses a strongly acidic or basic polyelectrolyte polymer and at least about 5.5.
A method comprising the use of polystyrenesulfonic acid, polyvinylsulfuric acid or polyvinylbenzylammonium chloride as a polyelectrolyte polymer in a composition comprising a buffer substance capable of maintaining pH and a pH indicator means.

JP-A-59-133207 discloses a reagent containing a weakly basic polyelectrolyte polymer neutralized with a strong organic acid and an indicator, wherein the polyelectrolyte polymer is polyethyleneimine, polyvinylamine, What is described is a polyaminostyrene or a copolymer of monomers constituting them.

JP-A-59-133208 describes a composition containing a weakly basic polyelectrolyte polymer in which at least one carboxyl group is present in the form of an ammonium salt and an indicator.

Japanese Patent Laid-Open No. 2-66451 discloses at least 1
Compositions containing one pH buffer but no polyelectrolyte polymer or containing at least one pH buffer and / or at least one complexing agent and further containing one pH indicator are described. Has been done.

Japanese Unexamined Patent Publication (Kokai) No. 3-100461 describes a method using a dye or a dye precursor whose solubility correlates with ion concentration.

All of these methods have been developed as methods for measuring the ionic strength or specific gravity of a liquid sample using a test piece and have been partially put into practical use, but Na, K, etc., which cause urine specific gravity, Medium, only specific components are measured, and it does not always exactly match the specific gravity, and when measuring samples of various specific gravity ranks, there is little change in color difference in color tone in the high specific gravity range. It has a common problem that it is difficult to judge with the naked eye. Further, all of these methods have a relatively slow coloration speed, and therefore, when attempting to optically and automatically determine a large number of samples, it takes a long time for measurement, which is not preferable.

[0012] Therefore, as many substances as possible, which have a large color difference change in color tone and constitute a factor of specific gravity, can be measured with the naked eye even in the high specific gravity range, and the calibration range is wide.
In addition, there is a long-felt demand for the emergence of a method for measuring the ionic strength or specific gravity of a liquid sample, which has a fast coloration speed and can easily and optically determine a large number of samples.

[0013]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above-mentioned present situation, and it is possible to detect a change in specific gravity due to Na, K, Ca, Mg, etc., and a large change in color difference in color tone, and to detect liquid samples such as urine. In the measurement of ionic strength and specific gravity, it is easy to determine visually, and the coloration rate is fast and a large number of samples can be easily optically optically determined, a reagent composition for measuring ionic strength or specific gravity of a liquid sample, And a test piece obtained by impregnating, coating or applying the reagent composition.

[0014]

The present invention includes (1) (i) one or more pH values.
A buffer, (ii) one or more pH indicators, and (ii
i) An invention of a reagent composition for measuring ionic strength or specific gravity of an aqueous liquid sample, which comprises one or more kinds of surfactants as a sensitizer. Further, the present invention is
(2) An invention of a method for measuring the ionic strength or specific gravity of an aqueous liquid sample by mixing the aqueous liquid sample with the above-mentioned reagent composition and detecting the variation of the pH of the solution and the variation of the dissociation degree of the pH indicator. is there. Furthermore, the present invention is (3) an invention of a test piece for measuring ionic strength or specific gravity of an aqueous liquid sample obtained by impregnating an absorbent carrier with the above-mentioned reagent composition, or coating or coating on a film. Furthermore, the present invention is (4) the test piece is immersed in an aqueous liquid sample to impregnate the sample, and then the sample is taken out and the pH of the test piece after a certain period of time (10 seconds to several minutes) is adjusted. It is an invention of a method for measuring ionic strength or specific gravity of an aqueous liquid sample by detecting fluctuations and fluctuations in the dissociation degree of a pH indicator. The present invention also provides (5) the ionic strength of the aqueous liquid sample by dropping the aqueous liquid sample on the test piece and detecting the change in the pH of the test piece after a certain period of time and the change in the dissociation degree of the pH indicator. Or, it is an invention of a method for measuring specific gravity.

To briefly describe the features of the present invention, the present invention uses an interface instead of a polyelectrolyte or a complex-forming agent in the conventional method for measuring ionic strength or specific gravity by detecting a change in pH. The use of activators as sensitizers dramatically improves the measurement of ionic strength and specific gravity.

That is, the present invention has not only the solubilizing and wetting effects of the surfactant but, surprisingly, the reverse action of suppressing the dissociation of the pH indicator, which is usually promoted by the addition of salt,
As a result, p accompanying the pH change of the buffer due to the addition of salt
It is an invention completed by the present inventors' finding that the absorbance (OD) change of the H indicator is increased.
That is, generally, the pH of a buffer solution comprising a buffering agent having a dissociation degree of 1.00 or less, more strictly 0.9 or less, rises and falls due to the enhancement of ionic strength. For example, in the case of a buffer solution of a combination of a weak acid and a strong base, the pH is lowered by the addition of a strong electrolyte, and in the case of a buffer solution of a combination of a strong acid and a weak base, the pH is raised by the addition of a strong electrolyte.

In addition, a normal pH indicator, particularly a triphenylmethane type pH indicator, promotes dissociation even at the same pH by increasing the ionic strength. Therefore, this pH change is measured with a pH indicator and the absorbance change value (△
When measuring the ionic strength or specific gravity from OD), if a buffer solution of a combination of a weak acid and a strong base is used, the pH decrease of the buffer solution and the dissociation promoting action of the pH indicator are offset, and the absorbance change value (△) lower than the actual value OD) or, in some cases, a negative absorbance change value (ΔOD), which is hardly practical.

However, when a surfactant is added to this, not only the dissociation promotion of the pH indicator is suppressed and the absorbance change value (ΔOD) corresponding to the pH change is obtained, but further,
The present inventors have found that they have a sensitizing action and give an absorbance change value (ΔOD) equal to or higher than the pH change value (ΔpH), and have completed the present invention.

The pH buffer according to the present invention is usually
pH range of 3.0 to 10.0, preferably pH 4.0
Used in the range of up to 9.0, the pH may be changed by the change in concentration due to the addition of a strong electrolyte such as chloride or sulfate such as sodium chloride and potassium chloride, and for example, Phosphate buffer, borate buffer, citrate buffer, tris (hydroxymethyl) aminomethane-maleic acid buffer [Tris-maleic acid buffer], bis (2
-Hydroxyethyl) iminotris (hydroxymethyl) methane buffer [bistris buffer] and the like.

When the pH buffer according to the present invention is used in combination with a phosphate, a bis-tris buffer or the like containing a weak acid such as boric acid or citric acid, or a salt thereof, a salt such as NaCl during pH adjustment. This is preferable because it can prevent the occurrence of

Examples of the acid used here include boric acid, citric acid, malonic acid, maleic acid, metaphosphoric acid, tartaric acid, oxalic acid, phosphoric acid, phthalic acid and the like.

Further, examples of these salts include, for example,
Examples thereof include sodium salt, potassium salt, lithium salt, and ammonium salt.

The pH indicator used in the present invention is required to be an indicator having a color change range within the pH fluctuation range of the pH buffer agent. In particular, the triphenylmethane system whose dissociation is promoted by the enhancement of ionic strength. The pH indicator is preferably used, and specific examples thereof include bromthymol blue, bromcresol purple, thymol blue, phenol red, chlorophenol red, and bromcresol green, and methyl red other than triphenylmethane type. And the like can be mentioned as preferable pH indicators. In addition, even if these indicators are used alone,
You may use together. For example, when bromthymol blue is used in combination with another pH indicator, the change in color tone can be increased and the color change range can be expanded, which is more practical and more preferable. Examples of other pH indicators that can be used in combination with bromthymol blue include, for example, pH indicators such as thymol blue and phenol red.

The surfactant used in the present invention includes:
Any substance having an action of suppressing the dissociation of the pH indicator promoted by the enhancement of ionic strength may be used, and any of an anionic surfactant, a cationic surfactant, an amphoteric surfactant and a nonionic surfactant may be used. Is also good.

Examples of the anionic surfactant include sodium dodecyl sulfate (SDS), lithium dodecyl sulfate, and sodium dodecylbenzenesulfonate (S).
DBS), higher alcohol sulfate ester salts such as sodium dodecanesulfonate, sodium diisooctylsulfosuccinate (SDOSS), sodium octyl sulfate, etc., and preferably neutral to alkaline (pH 6 to 6).
Used with the pH buffer of 10).

Examples of cationic surfactants include, for example, quaternary ammonium salts having one or more alkyl groups having 7 or more carbon atoms, specifically, myristyltrimethylammonium bromide (MTAB), bromide. Cetyl trimethyl ammonium (CTAB), octadecyl trimethyl ammonium chloride, dodecyl trimethyl ammonium chloride, cetyl trimethyl ammonium chloride, lauryl trimethyl ammonium chloride, etc. are mentioned. In addition to these, quaternary ammonium salts having a phenyl group, for example, benzalkonium chloride, tetradecyldimethylbenzylammonium chloride, octadecyldimethylbenzylammonium chloride, lauryldimethylbenzylammonium chloride, and quaternary ammonium having a pyridyl group. Salts such as laurylpyridinium chloride, cetylpyridinium chloride and stearylamidomethylpyridinium chloride can also be mentioned as preferable ones. These quaternary ammonium salts can be used at any pH. Even if the quaternary ammonium salt is composed of only a lower alkyl group having 6 or less carbon atoms, the effect of the present invention is not recognized, and therefore it cannot be used for the purpose of the present invention.

Examples of the amphoteric surfactant include alkyl betaines such as lauryl dimethyl betaine and stearyl betaine, and 2-lauryl lu N-carboxymethyl-N-hydroxylethyl imidazolinium betaine. These are preferable. Neutral range (pH 6
~ 8) pH buffering agent. Examples of the nonionic surfactant include polyoxyethylene alkyl phenyl ethers such as polyoxyethylene (8) octyl phenyl ether and polyoxyethylene (10) octyl phenyl ether, and polyoxyethylene (20) sorbitan mono. Examples include polyoxyethylene alkyl esters such as laurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan trioleate, and sorbitan monolaurate. , These are preferably used with a pH buffer in the neutral range (pH 6-8).

The reagent composition of the present invention comprises any one of the above.
It is characterized in that it comprises at least one pH buffer, at least one pH indicator, and at least one surfactant as a sensitizer, but these three components are more preferable. Examples of the combination include the combinations shown in Table 1 below.

[0029]

[Table 1]

The reagent composition of the present invention is most advantageous especially when it is used as a test piece, but it can be used in the form of powder or tablet in addition to the test solution.

For example, when the reagent composition of the present invention is used as a test solution (that is, in a liquid type), an aqueous liquid sample and the reagent composition of the present invention are mixed to change the pH of the solution and a pH indicator. It suffices to detect the change in the dissociation degree. pH
Of the pH indicator and the dissociation degree of the pH indicator
It can be easily detected by measuring OD).

When the reagent composition of the present invention is used as a test solution, the mixing ratio of the above three components is 100 w when the total amount of the three components is 100 w.
/ W%, the amount of (i) one or more pH buffers is usually 45 to 99.89 w / w%, preferably 9
5-99.8 w / w%, more preferably 98-99.7.
w / w%, (ii) as the amount of one or more pH indicators,
Usually 0.01 to 25 w / w%, preferably 0.02 to
0.5 w / w%, more preferably 0.03 to 0.3 w /
The amount of w%, (iii) one or more kinds of surfactants is usually 0.1 to 30.0 w / w%, preferably 0.2 to 3.
0 w / w%, and more preferably 0.3 to 1.5 w / w%.

When the reagent composition of the present invention is used as a test piece, the test piece obtained by impregnating, coating or applying the reagent composition of the present invention is immersed in an aqueous liquid sample to impregnate the sample, This may be taken out and the fluctuation of the pH of the test piece and the fluctuation of the dissociation degree of the pH indicator after a predetermined time may be detected. For the variation of pH and the variation of dissociation degree of pH indicator, compare the coloration of the test piece after a certain time with the color tone of the standard color chart prepared by performing the same operation using a solution of known ionic strength or known specific gravity. Thus, it is possible to detect the color of the test piece after a certain period of time (10 seconds to several minutes) by optically determining the reflectance with naked eyes.

As another method using the test piece, there is also a method of dropping an aqueous liquid sample on the test piece and measuring the coloration of the test piece with the naked eye or optically by reflectance.

When the reagent composition of the present invention is used as a test piece, the mixing ratio of the three components is (i) one or more p
The amount of the H buffer is usually 45 to 99.89 w / w
%, Preferably 60 to 98.5 w / w%, (ii) The amount of one or more pH indicators is usually 0.01 to 25 w /
w%, preferably 1 to 15 w / w%, and the amount of (iii) one or more surfactants is usually 0.1 to 30.0 w /
w%, preferably 0.3 to 20 w / w%. .

Further, for example, 30 cm as an absorbent carrier
^When the filter paper of ² is used, (i) the amount of one or more pH buffering agents is 1-120 mg (5-900 mM solution), preferably 5-75 mg (20-500 mM solution), ( ii) The amount of one or more pH indicators is 0.
3-1 mg (dry weight), preferably 0.4-0.8 mg
(Dry weight), (iii) the amount of one or more surfactants is 0.1 to 1 mg (dry weight), preferably 0.2 to
0.8 mg (dry weight) may be mentioned.

When the reagent composition of the present invention is used as a test piece, it can be used in the form of various test pieces. That is, each reagent and other reagents and binders according to the present invention, for example, polyvinylpyrrolidone, polyvinyl alcohol, polyethylene glycol, hydroxypropylmethyl cellulose, gum arabic, gelatin, ethyl cellulose or the like is water or a suitable organic solvent or these. An absorbent carrier is impregnated with the reagent solution according to the present invention prepared by dissolving it in the mixed solvent of 1 to several times, or coated or applied on the carrier film one to several times and then dried to obtain a test sheet. This can be cut into an appropriate size and fixed on an appropriate support to form a stick or a tape in any form.

Examples of the absorbent carrier to be impregnated with the reagent solution include porous carriers such as cellulose fiber filter paper, cloth, non-woven fabric, paper and synthetic fiber filter paper. Further, as the film for coating or applying the reagent solution, , Synthetic films, aluminum laminated films and the like.

Examples of the support for holding the absorbent carrier or film include polyvinyl chloride, polyester,
Examples include sheets of synthetic polymer compounds such as Teflon, polystyrene, polyvinyl acetal, polypropylene, and polyvinylidene chloride, and cardboard coated with these.

The size, size and the like of the absorbent carrier, film and support are not particularly limited, and it is sufficient to use those having sizes and sizes commonly used for this type of purpose. Further, regarding the method of adhering the absorbent carrier or the film to the support, a method of using an adhesive, a method of using an adhesive tape, etc., which are usually performed in this kind of field,
Either method may be used and no special method is required.

When the reagent composition of the present invention is used as a test solution or a test piece, p in the test solution or impregnating solution, the coating solution or the coating solution (hereinafter abbreviated as test solution or impregnating solution) is used.
The concentration of the H buffer is usually 1 to 900 mM, preferably 5 to 300 mM.

The concentration of the pH indicator in the test solution or impregnating solution is usually 0.0005 to 0.5 w / v%, preferably 0.001 to 0.2 w / v%.

Further, the concentration of the surfactant in the test solution or impregnating solution is usually 0.001 to 2.0 w / v%, preferably 0.005 to 1.0 w / v%.

Next, the present invention will be described more specifically with reference to the following examples and comparative examples, but the scope of the present invention is not limited thereby.

[0045]

【Example】

Example 1 Disodium hydrogen phosphate dodecahydrate 1.07 g Monosodium dihydrogen phosphate dihydrate 0.02 g Sodium dodecyl sulfate 0.07 g Bromthymol blue 0.10 g Distilled water 100 ml Reagent having the above composition A solution was prepared, and a filter paper for chromatography (30 cm ² , the same applies hereinafter) was immersed in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The prepared test sheet was cut into a rectangle of 5 × 6 mm sides and attached to a polyvinyl chloride sheet (6 mm × 11 cm) using a double-sided tape to give a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM40
5. The reflectance value using Wako Pure Chemical Industries, Ltd. showed a proportional relationship with each specific gravity rank value after 30 seconds. Also,
The coloring was highly sensitive, and the effect of increasing the sensitivity by the surfactant was recognized.

[0046]

Example 2 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Sodium diisooctyl sulfosuccinate 0.1 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml A reagent solution having the above composition was prepared, a filter paper for chromatography was dipped in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. In addition, as in Example 1, high-sensitivity coloration was exhibited, and the sensitivity increasing effect of the surfactant was recognized.

[0048]

Comparative Example 1 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml Reagent solution of the above composition (ie, Example) 2) was prepared by removing the surfactant), chromatographic filter paper was immersed in the composition to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds, but the coloration sensitivity was lower than that in Example 2.

[0050]

Example 3 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Sodium dodecyl sulfate 0.07 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml Reagents having the above composition A solution was prepared, and a chromatographic filter paper was immersed in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. Further, as in Examples 1 and 2, highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0052]

Example 4 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Sodium dodecylbenzenesulfonate 0.09 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml The above composition Of the reagent solution was prepared, and the filter paper for chromatography was immersed in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. Further, similarly to Examples 1 to 3, coloration with high sensitivity was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0054]

Example 5 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Sorbitan monolaurate 0.10 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml A reagent solution was prepared, a filter paper for chromatography was dipped in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows a different color when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. In addition, as in Examples 1 to 4, a highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0056]

Example 6 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Polyoxyethylene (10) octyl phenyl ether 0.10 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distillation 100 ml of water A reagent solution having the above composition was prepared, and a filter paper for chromatography was immersed in this to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. Further, as in Examples 1 to 5, highly sensitive coloration was shown, and the effect of increasing the sensitivity by the surfactant was recognized.

[0058]

Example 7 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Polyoxyethylene (20) sorbitan monolaurate 0.05 g Bromthymol blue 0.09 g Thymol blue 0.04 g Steam Distilled water 100 ml A reagent solution having the above composition was prepared, and a chromatographic filter paper was immersed in this to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. In addition, as in Examples 1 to 6, highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0060]

Example 8 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Lauryl dimethyl betaine 0.10 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml Reagents having the above composition A solution was prepared, and a chromatographic filter paper was immersed in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows a different color when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. Further, similarly to Examples 1 to 7, highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0062]

Example 9 Polyvinylpyrrolidone (K-30) 40 g Disodium hydrogen phosphate dodecahydrate 1.07 g Folic acid 0.09 g Sodium dodecyl sulfate 0.07 g Bromthymol blue 0.09 g Thymol blue 0.04 g 100 ml of distilled water (adjusted to pH 7.75 with 20% NaOH solution) A reagent solution having the above composition was prepared, applied on a plastic film, and then dried at 55 to 65 ° C. The prepared test sheet was cut into a rectangle of 5 × 6 mm sides and attached to a polyvinyl chloride sheet (6 mm × 11 cm) using a double-sided tape to give a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. In addition, as in Examples 1 to 8, highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0064]

Example 10 Monosodium dihydrogen phosphate dihydrate 3.51 g Fouic acid 0.70 g Sodium hydroxide 0.75 g Sodium diisooctyl sulfosuccinate 0.05 g Polyethylene glycol-4000 0.05 g Brom Thymol blue 0.09 g Thymol blue 0.04 g Ethyl alcohol 12.5 ml Distilled water 87.5 ml Prepare a reagent solution of the above composition, and dip a chromatographic filter paper in it to impregnate the reagent with 55-65 ° C. Dried. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. Further, as in Examples 1 to 9, high-sensitivity coloration was shown, and the sensitivity increasing effect of the surfactant was recognized.

[0066]

Example 11 Disodium hydrogen phosphate dodecahydrate 1.07 g Monosodium dihydrogen phosphate dihydrate 0.02 g Sodium dodecyl sulfate 0.07 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml A reagent solution having the above composition was prepared, and a filter paper for chromatography was immersed in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. In addition, as in Examples 1 to 10, highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0068]

Example 12 Disodium hydrogen phosphate dodecahydrate 1.07 g Monosodium dihydrogen phosphate dihydrate 0.02 g Sodium dodecyl sulphate 0.07 g Bromthymol blue 0.05 g Phenol red 0.05 g Distilled water 100 ml A reagent solution having the above composition was prepared, and a filter paper for chromatography was immersed in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. In addition, as in Examples 1 to 11, highly sensitive coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0070]

Example 13 Citric acid monohydrate 1.05 g Sodium hydroxide 0.2 g Myristyltrimethylammonium bromide 0.05 g Bromcresol green 0.1 g Distilled water 100 ml (adjusted to pH 4.0). ) A reagent solution having the above composition was prepared, a filter paper for chromatography was dipped in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. This test piece shows different coloration when immersed in each of the following specific gravity urine samples measured with a pycnometer, and a reflectance measuring device (Pretester RM405, manufactured by Wako Pure Chemical Industries, Ltd.) is used. The reflectance value showed a proportional relationship with each specific gravity rank value after 30 seconds. Further, as in Examples 1 to 12, high-sensitivity coloration was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0072]

Comparative Example 2 Citric acid monohydrate 1.05 g Sodium hydroxide 0.2 g Bromcresol green 0.1 g Distilled water 100 ml (adjusted to pH 4.0) Reagent solution of the above composition (ie, The composition of Example 13 from which the surfactant was removed was prepared, and a filter paper for chromatography was dipped in the composition to impregnate it with a reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 m
(m × 11 cm) to give a test piece. This test piece is
When immersed in each of the following specific gravity urine samples measured with a pycnometer, they show different coloration and reflectivity measurement device (Pretester RM405, Wako Pure Chemical Industries, Ltd.)
The reflectance value using was proportional to each specific gravity rank value after 30 seconds, but the coloration sensitivity was lower than that in Example 13.

[0074]

Example 14 Tris (hydroxymethyl) aminomethane 2.42 g Maleic acid 2.32 g Sodium hydroxide 0.79 g Sodium dodecyl sulfate 0.02 g Bromthymol blue 2 mg Distilled water 100 ml A reagent solution having the above composition was prepared, 0.4 ml of each of the following specific gravity urine samples measured with a pycnometer was mixed with 5 ml thereof, and then the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
The sensitivity increasing effect of the surfactant was confirmed.

[0076]

Comparative Example 3 Tris (hydroxymethyl) aminomethane 2.42 g Maleic acid 2.32 g Sodium hydroxide 0.79 g Bromthymol blue 2 mg Distilled water 100 ml A reagent solution having the above composition (ie, from the composition of Example 14 to the interface 0.4 ml of the following specific gravity urine samples measured with a pycnometer for 5 ml of the
After mixing, the absorbance change was measured using a spectrophotometer (Hitachi 557 type). As shown below, the measured values showed almost no proportional relationship with each specific gravity rank value, and showed extremely low sensitivity as compared with Example 14.

[0078]

Example 15 Monosodium dihydrogen phosphate dihydrate 2.61 g Fouic acid 1.03 g Sodium hydroxide 0.50 g Sodium dodecyl sulfate 0.02 g Bromthymol blue 2 mg Distilled water 100 ml Reagents having the above composition A solution was prepared, and 0.4 ml of each of the following specific gravity urine samples measured by a pycnometer was mixed with 5 ml of the solution, and the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to the case of the test piece, the effect of increasing the sensitivity by the surfactant was recognized.

[0080]

Comparative Example 4 Monosodium dihydrogen phosphate dihydrate 2.61 g Folic acid 1.03 g Sodium hydroxide 0.50 g Bromthymol blue 2 mg Distilled water 100 ml Reagent solution of the above composition (ie, Example) 15 ml of the composition (excluding surfactant) was prepared, and 5 ml thereof was analyzed with a pycnometer.
After mixing, the absorbance change was measured using a spectrophotometer (Hitachi 557 type). As shown below, the measured values showed almost no proportional relationship with each specific gravity rank value, and showed extremely low sensitivity as compared with Example 15.

[0082]

Example 16 Citric acid monohydrate 1.41 g Sodium hydroxide 0.52 g Myristyltrimethylammonium bromide 0.02 g Chlorphenol red 5 mg Distilled water 100 ml A reagent solution having the above composition was prepared, and 5 ml thereof was prepared. On the other hand, 0.4 ml of the following specific gravity urine samples measured with a pycnometer were mixed, and then the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Example 15, it showed high sensitivity, and the effect of increasing the sensitivity by the surfactant was recognized.

[0084]

Example 17 Boric acid 1.24 g Potassium chloride 1.49 g Sodium hydroxide 0.37 g Myristyltrimethylammonium bromide 0.02 g Thymol blue 2.5 mg Distilled water 100 ml A reagent solution having the above composition was prepared, and 5 ml thereof was prepared. On the other hand, after mixing 0.4 ml of each of the following specific gravity urine samples measured with a pycnometer, the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Examples 15 to 16, it showed high sensitivity, and the effect of increasing the sensitivity by the surfactant was recognized.

[0086]

Example 18 Monosodium dihydrogen phosphate dihydrate 0.54 g Disodium hydrogen phosphate dodecahydrate 2.35 g Polyoxyethylene (10) octyl phenyl ether 12.5 mg Bromthymol blue 4 mg Steam Distilled water 100 ml Prepare a reagent solution having the above composition, mix 5 ml of each with the following specific gravity urine sample 0.4 ml measured with a pycnometer, and then measure the change in absorbance using a spectrophotometer (Hitachi 557 type). It was measured. The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Examples 15 to 17, high sensitivity was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0088]

Example 19 Monosodium dihydrogen phosphate dihydrate 0.54 g Disodium hydrogen phosphate dodecahydrate 2.35 g Cetyltrimethylammonium bromide 0.02 g Bromthymol blue 3.8 mg Distilled water 100 ml A reagent solution having the above composition was prepared, and 0.4 ml of each of the following specific gravity urine samples measured by a pycnometer was mixed with 5 ml thereof, and then the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). .. The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Examples 15 to 18, it showed high sensitivity, and the effect of increasing the sensitivity by the surfactant was recognized.

[0090]

Comparative Example 5 Monosodium dihydrogen phosphate dihydrate 0.54 g Disodium hydrogen phosphate dodecahydrate 2.35 g Tetrabutylammonium bromide 20 mg Bromthymol blue 3.8 mg Distilled water 100 ml Above Instead of the reagent solution having the composition [that is, cetyltrimethylammonium bromide (a quaternary ammonium salt having one alkyl group having 16 carbon atoms and three alkyl groups having 1 carbon atom) in Example 19] tetrabutyl bromide Ammonium (having 4 alkyl groups each having 4 carbon atoms) was prepared], and 0.4 ml of each of the following specific gravity urine samples measured by a pycnometer was mixed with 5 ml thereof, and then the spectrophotometer ( The change in absorbance was measured using a Hitachi 557 model. The results are shown below, and the sensitivity was remarkably lower than that of Example 19.

[0092]

In the above, instead of tetrabutylammonium bromide, a quaternary ammonium water used as a neutralizing agent for the polymer electrolyte polymer in JP-A-59-133208. The same result as above was obtained when the oxide tetrabutylammonium hydroxide was used.

Example 20 Vise (2-human oxyethyl) iminotris (human oxymethyl) methane 3.94 g boric acid 0.07 g lauryl dimethyl betaine 0.02 g bromothymol blue 4 mg distilled water 100 ml A reagent solution having the above composition was prepared, 0.4 ml of each of the following specific gravity urine samples measured with a pycnometer was mixed with 5 ml thereof, and then the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Examples 15 to 19, it showed high sensitivity, and the effect of increasing the sensitivity by the surfactant was recognized.

[0095]

Example 21 Monosodium dihydrogen phosphate dihydrate 0.54 g Disodium hydrogen phosphate dodecahydrate 2.35 g Benzalkonium chloride 40 mg Bromthymol blue 3.8 mg Distilled water 100 ml The above composition The reagent solution was prepared and mixed with 0.4 ml of the following specific gravity urine samples measured with a pycnometer, and the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Examples 15 to 20, it showed high sensitivity, and the effect of increasing the sensitivity by the surfactant was recognized.

[0097]

Example 22 Monosodium dihydrogen phosphate dihydrate 0.54 g Disodium hydrogen phosphate dodecahydrate 2.35 g Cetylpyridinium chloride 10 mg Bromthymol blue 2.3 mg Distilled water 100 ml With the above composition A reagent solution was prepared, and 0.4 ml of each of the following specific gravity urine samples measured with a pycnometer was mixed with 5 ml of the reagent solution, and then the change in absorbance was measured using a spectrophotometer (Hitachi 557 type). The measured value showed a proportional relationship with each specific gravity rank value as follows. Also, the change in absorbance is large,
Similar to Examples 15 to 21, high sensitivity was exhibited, and the effect of increasing the sensitivity by the surfactant was recognized.

[0099]

Example 23 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Sodium diisooctyl sulfosuccinate 0.1 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml A reagent solution having the above composition was prepared, a filter paper for chromatography was dipped in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. When the test pieces were immersed in the following aqueous NaCl solutions having different concentrations, the test pieces showed different colors, and the reflectance measuring device (Pretester RM4
05, the reflectance value using Wako Pure Chemical Industries, Ltd. is 30
After a second, each concentration rank value, that is, a proportional relationship with the ionic strength was shown. In addition, all the colors were highly sensitive, and the effect of increasing the sensitivity by the surfactant was recognized.

[0101]

Comparative Example 6 Disodium hydrogen phosphate dodecahydrate 1.07 g Houlic acid 0.09 g Bromthymol blue 0.09 g Thymol blue 0.04 g Distilled water 100 ml Reagent solution of the above composition (ie, Example) 23) was prepared by removing the surface active agent from the composition of Example 23, and a filter paper for chromatography was immersed in the composition to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 m
(m × 11 cm) to give a test piece. This test piece is
When immersed in the following NaCl aqueous solutions with different concentrations, they show different colors and reflectivity measuring device (Pretester RM
405, the reflectance value using Wako Pure Chemical Industries, Ltd. is 3
After 0 seconds, each density rank value, that is, the proportional relationship with the ionic strength was shown, but the coloration sensitivity was lower than that in Example 23.

[0103]

Example 24 Monosodium dihydrogen phosphate dihydrate 0.02 g Disodium hydrogen phosphate dodecahydrate 1.07 g Sodium dodecyl sulfate 0.07 g Bromthymol blue 0.10 g Distilled water 100 ml Above A reagent solution having the composition was prepared, a filter paper for chromatography was dipped in the solution to impregnate the reagent, and then dried at 55 to 65 ° C. The produced test sheet is cut into a rectangle of 5 × 6 mm side, and a polyvinyl chloride sheet (6 mm
X 11 cm) and used as a test piece. When the test pieces were immersed in the following aqueous NaCl solutions having different concentrations, the test pieces showed different colors, and the reflectance measuring device (Pretester RM4
05, the reflectance value using Wako Pure Chemical Industries, Ltd. is 30
After a second, each concentration rank value, that is, a proportional relationship with the ionic strength was shown. Further, similarly to Example 23, the coloration was highly sensitive, and the effect of increasing the sensitivity by the surfactant was recognized.

[0105]

As is clear from the above results, all the reagent compositions of Examples 1 to 24 of the present invention showed highly sensitive color development. On the other hand, Comparative Examples 1 to 6 all showed low values of color sensitivity as compared with the reagent compositions of Examples 1 to 24.

[0107]

The present invention provides an ionic strength or specific gravity of an aqueous liquid sample which comprises one or more pH buffering agents, one or more pH indicators, and one or more surfactants as sensitizers. The present invention provides a reagent composition for measurement, which is a problem when using a conventional reagent composition for measuring ionic strength or specific gravity by using the reagent composition of the present invention, that is, there is little change in absorbance and reproducibility. Is poor, there is little change in color difference in color tone, it is difficult to determine with naked eyes when measuring the ionic strength and specific gravity of liquid samples such as urine, and because the coloration speed is slow, many samples are automatically This is an invention that has a remarkable effect in that it takes a long time to make a determination and can solve all the disadvantages and the like, and is a great invention that contributes to the related art.

Claims (9)

[Claims] 1. (i) one or more pH buffering agents, (ii) 1
One or more pH indicators, and (iii) 1 as a sensitizer
One or more surfactants, characterized in that
A reagent composition for measuring ionic strength or specific gravity of an aqueous liquid sample. 2. The pH buffering agent has a pH of 3.0 to 10.0.
The reagent composition according to claim 1, which is a buffer used in the range. 3. The ionic strength or specific gravity of the aqueous liquid sample is obtained by mixing the aqueous liquid sample with the reagent composition according to claim 1 and detecting the variation of the pH of the solution and the variation of the dissociation degree of the pH indicator. How to measure. 4. The method according to claim 3, wherein changes in pH and changes in the dissociation degree of the pH indicator are detected by measuring changes in absorbance. 5. A test piece for measuring the ionic strength or specific gravity of an aqueous liquid sample, which is obtained by impregnating an absorbent carrier with the reagent composition according to claim 1 or coating or applying it on a film. 6. The test piece according to claim 5 is immersed in an aqueous liquid sample to impregnate the sample, and then the sample is taken out, and the pH fluctuation of the test piece after a certain period of time and the dissociation degree of the pH indicator are measured. A method for measuring the ionic strength or specific gravity of an aqueous liquid sample by detecting fluctuations. 7. A method for visually observing the color change of the test piece after a certain period of time for the change in pH and the change in the dissociation degree of the pH indicator,
Alternatively, the method according to claim 7, wherein the detection is performed by optically measuring the reflectance. 8. An aqueous liquid sample is dropped on the test piece according to claim 5, and after a certain period of time, fluctuations in pH of the test piece and p
A method for measuring the ionic strength or specific gravity of an aqueous liquid sample by detecting a change in the dissociation degree of the H indicator. 9. A method for visually observing the color change of the test piece after a certain period of time for the change in pH and the change in the dissociation degree of the pH indicator,
Alternatively, the method according to claim 8, wherein the detection is performed by optically measuring the reflectance. [0001]