JP3014633B2 - Urine test method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for testing urine collected from humans and other animals. In particular, it is used for testing the concentration of sugar, protein, etc. in urine.

[0002]

2. Description of the Related Art A healthy adult usually has a daily dose of 1,000.
Drain ~ 1500 ml of urine. Among them, the total solid content is 5
0-70 g. About 25g of the solid content,
It is an inorganic substance mainly composed of sodium chloride, potassium chloride and phosphoric acid, and is mostly dissolved in an ionized state. The rest is organic, mainly urea and uric acid, but slightly
Sugar and protein also exist. Conventionally, as a method for testing such sugars and proteins in urine, a test paper or the like impregnated with a reagent is immersed in urine, and the color reaction thereof is observed with a spectrometer or the like.

[0003]

However, the test papers used here are required to be different from each other in accordance with individual test items such as sugar and protein. Also,
Since a new test paper is required for each inspection, there is a drawback that running costs are increased. Further, there is a limit to automation for labor saving. In addition, when used at home, a layman is required to set and exchange test papers. This work was relatively hated, which hindered the spread of urine testing devices to homes. An object of the present invention is to provide a urine test method that can be easily maintained and managed without using consumables such as test papers in consideration of these problems.

[0004]

The urine test method of the present invention comprises:
Normal urine protein concentration (generally less than 10 mg / dl)
By measuring the optical rotation angle of the urine such, Guru of the urine
This is to determine the density of the course . Sugar, ie, glucose, contained in urine is usually 0.13 per day.
~ 0.5g is excreted in urine. From this and the amount of urine, the concentration, that is, the urine sugar value was roughly estimated, and on average, about 50 mg /
dl or less. This urine sugar level reflects a part of the health condition. In particular, in the case of diabetes, this is several hundred mg /
dl, and in some cases can reach several thousand mg / dl. That is, it may be against the normal value, in degrees, up <br/> pressurized takes about one to two orders of magnitude. On the other hand, protein contained in urine, that is, albumin is usually less than glucose,
It is excreted in urine at 3-60 mg / dl . Estimated from urine volume, the concentration in urine is on average about 6 mg / dl.
It is as follows. This albumin concentration also reflects part of the state of health, and in the presence of kidney disease, the albumin concentration will be 100 mg / dl or more. That is, it may increase by about one digit or more with respect to the normal value.

[0005] Glucose and protein present in urine show optical rotation, whereas urea and uric acid, which are the main components of organic matter in urine, do not show optical rotation. Also, all minerals in urine do not show optical rotation. Therefore, by measuring the optical rotation angle of urine, the concentration of glucose or protein in urine can be accurately determined. Similarly, even when urine contains L-ascorbic acid (so-called vitamin C), it is possible to determine the L-ascorbic acid concentration by measuring the angle of rotation.

Hereinafter, the measurement principle of the present invention will be described. The angle of rotation A is proportional to the product of the specific rotation α of the optically rotating substance and the concentration C. This relationship is shown in equation (1). When one kind of optically active substance is used, A [deg] = L [cm] × α × C [kg / dl] (1), and when N kinds of optically active substances are included, A [deg] = L × (α ₁ × C ₁ + α ₂ × C ₂ +... + Α _N × C _N ) (2) Here, L is a measurement optical path length. Table 1 shows the specific rotation α of glucose and albumin.

[0007]

[Table 1]

The specific rotation shown here is that of an aqueous glucose solution and an aqueous albumin solution at 20 ° C. That is, in an aqueous glucose solution having a concentration of 100 g / dl, a wavelength of 5
When light of 89 nm propagates for 10 cm, the polarization direction is 5
Turn 0 deg. However, in practice, the above concentration cannot be realized due to the limitation of the solubility of glucose, but (1)
According to the equation, since the angle of rotation and the concentration are proportional, 100 mg / d
In l, 50 × 10 ^-3 to deg rotated. From these, when the optical rotation substance in urine is only glucose, the urinary sugar value can be calculated from the specific rotation of glucose by measuring the optical rotation angle of urine. Similarly, albumin and L
-It can also be calculated for ascorbic acid. When a plurality of optically active substances such as glucose, albumin and L-ascorbic acid coexist in urine, the respective concentrations can be calculated by the following method.

In addition, the optical rotation angle of urine having a known optical rotation angle range expressed by an interfering optical rotation substance other than the optical rotation substance whose concentration is unknown is measured, and the concentration of the optical rotation substance is determined by (A-A _h ) / ( α × L) ≦ C ≦ (A−A _l ) / (α × L) (3) where, A: the measured optical rotation angle of urine [deg] A _h : the maximum optical rotation angle developed by the interfering optical rotation substance Value [deg] _Al : Minimum value of the optical rotation angle developed by the interfering optically rotating substance [deg] α: Specific rotation of the optically rotating substance [deg / cm · dl / k]
g] L: Determined to be within the range of the measured optical path length [cm]. First, the equation (2) is modified to obtain the following equation (4). A = A _x + A _d (4) where A _x = L × α ₁ × C _{1 Ad} = L × (α ₂ × C ₂ +... + Α _N × C _N ). In (2), of optical rotation to be detected substance 1 substance X, optical active substance other than that material 2 to N, i.e. the interfering optical activity material, A _d is the angle of rotation expressed by interfering optical rotation material Equivalent to. Here, if a known concentration range of material 2 to N, the maximum value A _h and the minimum value A _l can take the A _d is known. From this, the following equation (5) is derived. A-A _h ≤A _x = A-A _d ≤A-A _l (5) The range of the optical rotation angle A _x is known from the equation (5), and the range of the concentration C _x is obtained from the specific rotation α _x and the measured optical path length L. I understand. This (5)
The expression (3) expresses the concentration C of the optical rotatory substance X in the expression. From this, for example, in the case of inspecting the glucose concentration, when the concentration of albumin which causes interference is 10 mg / dl or less, that is, when the minimum value of the albumin concentration is 0 and the maximum value is 10 mg / dl, the wavelength is 589 nm. In the measurement optical path length = 10 cm, A _h
= 0 deg, the ^{_{A l = -6 × 10 -3 deg}} . At this time, A =
Assuming that the measurement is 0.1 deg, from Table 1, the glucose concentration C (mg / dl) can be determined to be 200 ≦ C ≦ 212. In fact, when the urinary glucose level shows an abnormality, it is often sufficient if the test can be performed with the above accuracy, considering that the urine sugar level reaches several hundred mg / dl or more. That is, when the albumin concentration is equal to or less than the normal value of about 10 mg / dl, the presence or absence of abnormalities in the urinary sugar level can be examined by measuring the optical rotation angle at one wavelength.

[0010] Furthermore, the concentration of the optically rotating substance in the urine is determined by measuring the angle of optical rotation of urine containing N kinds of optically rotating substances using light of at least N kinds of wavelengths. The specific rotation differs for each wavelength depending on the optical rotation dispersion. Therefore, when N kinds of optically rotating substances coexist, by measuring the optical rotation angle at N kinds of wavelengths,
By using the above equation (2), N simultaneous equations independent of each other are obtained. From this, the concentrations of the N types of optically active substances can be calculated. Thus, by measuring the angle of rotation of urine, abnormalities in urine sugar level and albumin concentration can be inspected.

[0011] In addition, the optical rotation angle of urine with respect to light having a wavelength of 500 nm or more is measured. If the wavelength is shorter than 500 nm, the absorption mainly by urochrome (yellow component contained in urine) is increased, which may rather deteriorate the measurement accuracy.

Furthermore, the concentration of the light scattering substance in the urine is determined by measuring the amount of scattered light in the urine. Further, the light scattering substance is at least one of a protein and blood. Albumin, a protein in urine, has a molecular weight of about 70,000, and the scattering of light by this is smaller than the scattering of light by other organic and inorganic molecular weight substances in urine such as glucose (molecular weight: about 180). Big enough. That is, the scattering of light propagating in urine is mainly caused by albumin. Therefore, the albumin concentration can be determined by irradiating the urine with light and observing the scattered light directly or in the form of a decrease in the amount of transmitted light. It is also possible to test for the presence of blood, which is relatively large particles.

In addition, the amount of scattered light of urine with respect to light having a wavelength of 500 nm or more is measured. This is because, similarly to the measurement of the specific rotation, when the wavelength is shorter than 500 nm, the absorption mainly by urochrome is increased, which may deteriorate the measurement accuracy.

Further, by simultaneously measuring the optical rotation angle and the scattered light amount of urine, the concentrations of the optical rotation substance and the light scattering substance are simultaneously determined. In particular, it is effective when glucose and albumin are present in urine to the extent that they cannot be ignored.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION

[Embodiment 1] The first embodiment will be described in detail below. FIG. 1 is a configuration diagram of the polarimeter used in the present embodiment. The basic principle of this polarimeter is an optical null method based on polarization plane vibration using the Faraday effect. The 180 W low-pressure sodium lamp 1 emits parallel light. The bandpass filter 2 transmits only light having a wavelength of 589.0 nm among the light emitted from the sodium lamp 1. Polarizer 3
Transmits, for example, only light having a polarization component parallel to the paper surface of the light from the bandpass filter 2. With the modulation signal current input from the Faraday cell driver 5, the Faraday cell 4 modulates the polarization direction of the transmitted light to a small width by the optical Faraday effect. The substantial optical path length of the sample cell 6 containing urine is 10 cm. Rotating analyzer 7
Can be set to an arbitrary angle by the analyzer driver 8. Reference numeral 9 denotes an optical sensor that outputs a signal based on the intensity of transmitted light. The lock-in amplifier 10 performs phase-sensitive detection of the output of the optical sensor 9 using the modulated signal to the Faraday cell 4 as a reference signal. The computer 11 records the output of the lock-in amplifier 10 while sending a command to continuously rotate the rotary analyzer 7 to the analyzer driver 8. Thus, the function of finding the angle of the rotary analyzer 7 at which the output of the lock-in amplifier 10 becomes zero and calculating the optical rotation angle is provided. With the above configuration, an accuracy of about 10 ⁻³ deg is achieved.

The urinalysis in this example was performed as follows. First, a calibration curve is created to confirm the characteristics of the polarimeter. Pure water is put into the cell 6, and the lock-in amplifier 10
The angle of the rotary analyzer 7 at which the output of the sample becomes zero was measured. Using this angle as a reference, the optical rotation angle of a glucose aqueous solution prepared using pure water as a solvent and having concentrations of 20, 100, 200, 300, and 500 mg / dl, respectively, was measured. This result is shown by a white circle in FIG. From this, it was demonstrated that the glucose concentration could be measured. Next, the glucose concentration was previously adjusted to 50 mg / dl by a urine analyzer.
Hereinafter, the optical rotation angle of urine whose albumin concentration was determined to be 10 mg / dl or less was measured. Further, using this urine as a solvent, the concentration is 20, 100, 200, 300, 500 m, respectively.
g / dl glucose solution, ie, artificially formulated with diabetes. These optical rotation angles were measured. The results are shown by black circles in FIG. Since the optical rotation angle (black circle) of this artificial diabetes is represented by a straight line shifted 1.5 × 10 ⁻² deg parallel from the calibration curve, it accurately reflects the glucose concentration. The optical rotation angle of this urine alone was 1.5 × 10 ⁻² deg. From this and the albumin concentration range previously obtained by the urine analyzer, the glucose concentration C (mg / dl) can be determined to be in the following range by the equation (3). 30 ≦ C ≦ 42 This is in agreement with the result analyzed in advance.

Further, the urine analyzer has a glucose concentration of 300 mg / dl or more and an albumin concentration of 10 mg / dl.
For the urine determined to be / dl or less, the optical rotation angle was measured in the same manner. As a result, the optical rotation angle of this urine was 2.2 × 10 ⁻¹ de
g, and from the albumin concentration range obtained in advance by the urine analyzer, the glucose concentration C (mg / d
1) can be determined to be in the following range. 440 ≦ C ≦ 452 This also agrees with the result analyzed in advance. Thus, the albumin concentration was normal, that is, 10 mg / dl.
In the case of urine as described below, the measurement of the optical rotation angle allows the abnormality in the glucose concentration in urine (urine sugar level) to be detected with high accuracy. For example, a glucose concentration of 300 mg / dl or more can be determined with an error of about 12 mg / dl. As described above, according to the present embodiment, the urine sugar level can be inspected without using consumables such as test papers, and the practical effect is extremely large.

[Embodiment 2] This embodiment is an example of a urinalysis method in a case where glucose and albumin are present in urine to an extent not negligible. This embodiment will be described in detail below with reference to FIG. FIG. 3 is a configuration diagram of the polarimeter used in this embodiment. The basic principle of the polarimeter is the optical null method based on the polarization plane vibration using the Faraday effect, as in the first embodiment, and uses a light source having a different wavelength from the light source. Therefore, 3-11
The steps up to this are the same as those used in the first embodiment. 12
Is a semiconductor laser, which emits parallel light having an emission wavelength of 830 nm and 5 mW. This polarimeter operated in the same manner as in Example 1, and also achieved an accuracy of about 10 ^-3 deg. In this embodiment, the polarimeter used in the first embodiment is also used.

First, a calibration curve is prepared to confirm the characteristics of the polarimeter. Pure water was put into the cell, and the angle of the rotary analyzer 7 at which the output of the lock-in amplifier 10 became zero was measured. On the other hand, the concentrations were 20, 100, 200, 30 respectively.
A 0,500 mg / dl aqueous glucose solution and an albumin aqueous solution having a concentration of 20, 50, 100 mg / dl, respectively, were prepared. These optical rotation angles were measured with reference to the angle of the analyzer in the case of pure water. The result was as shown in FIG. 4, and it was confirmed that concentration measurement was possible.
The same sample was measured using a polarimeter using the light source having a wavelength of 589 nm used in Example 1. The result is shown in FIG.

From the equation (2) and the specific rotation shown in Table 1, the simultaneous equations shown in the equations (6) and (7) are established. Assuming that the concentrations of glucose and albumin are C ₁ and C ₂ (both in [kg / dl]), A ₅₈₉ = 0.1 × (50C ₁ -60C ₂ ) (6) A ₈₃₀ = 0.1 × (25C ₁₎ −10C ₂ ) (7) Here, A ₅₈₉ : the optical rotation angle [deg] for the light having the wavelength of 589 nm A ₈₃₀ : the optical rotation angle [deg] for the light having the wavelength of 830 nm. When the above A ₅₈₉ and A ₈₃₀ are measured, two unknowns C ₁ and C ₂ can be calculated from the equations (5) and (6). In fact, 100 ml of albumin per dl
g, glucose and 300 mg of coexisting aqueous solution were prepared, and the optical rotation angle was measured in the same manner. As a result, A ₅₈₉ = 9 × 10 ^-2 [deg] A ₈₃₀ = 6.5 × 10 ^-2 [deg] is obtained, and this is used to solve the simultaneous equations of equations (6) and (7). , It was confirmed that the ratio was the same as the charge ratio.

Similarly, when the glucose concentration is 50 mg / dl
Hereinafter, the optical rotation angle of urine determined to have an albumin concentration of 100 mg / dl or more was measured. As a result, a wavelength of 589 nm
The optical rotation angle A _{589 at} the time of and the optical rotation angle A _{830 at the} wavelength of 830 nm were A ₅₈₉ = −6 × 10 ⁻² [deg] A ₈₃₀ = −5 × 10 ⁻³ [deg], respectively. By solving the equations (5) and (6) using these, the glucose concentration was 30 mg / dl and the albumin concentration was 125 mg / dl, which were consistent with the analysis results.

The glucose concentration is 300 mg / dl.
As described above, the optical rotation angle of urine whose albumin concentration was determined to be 100 mg / dl or more was measured. As a result, a wavelength of 589 nm
The optical rotation angle A _{589 at} the time of and the optical rotation angle A _{830 at the} wavelength of 830 nm were A ₅₈₉ = 1 × 10 ⁻¹ [deg] and A ₈₃₀ = 8 × 10 ⁻² [deg], respectively. By solving equations (5) and (6) using these, the glucose concentration = 380 mg / dl,
An albumin concentration of 150 mg / dl was obtained, which was consistent with the analysis results. As described above, according to the present embodiment, even when glucose and albumin are present in urine to the extent that they cannot be ignored, the urinary sugar level and albumin concentration are inspected without using consumables such as test papers. It is possible,
Its practical effect is extremely large.

[Embodiment 3] This embodiment relates to a method for examining urinary albumin concentration and hematuria by light scattering. The configuration of the present embodiment will be described below with reference to FIG. The helium neon laser 13 has a wavelength of 63
3 nm, 5 mW parallel light is emitted. The substantial optical path length of the sample cell 14 containing urine is 10 cm and the width is 1 cm. Since the side surface of the sample cell 14 is also transparent, scattered light in the direction perpendicular to the optical path can be transmitted to the outside of the sample cell 14. The optical sensor 15 is arranged so that its viewing angle matches the sample cell 14, and can detect a scattered component of the laser light propagating in urine. Concentration 2
0, 50, and 100 mg / dl aqueous albumin solutions were prepared respectively, and the amount of scattered light from these and pure water was measured. The result was as shown in FIG. 7, and it was confirmed that there was a positive correlation between the albumin concentration and the amount of scattered light.

Next, a conventional urine analyzer has a glucose concentration of 50 mg / dl or less and an albumin concentration of 10 m / dl.
The amount of scattered light was measured for urine determined to be not more than g / dl, but no scattered light could be detected. The glucose concentration is 300 mg / dl or more, and the albumin concentration is 100 m / dl.
FIG. 7 shows a measurement of urine determined to be g / dl or more.
On the scale of で, about 6 signals were confirmed. Furthermore, about 6 signals could be confirmed by measuring urine having a glucose concentration of 50 mg / dl or less and an albumin concentration of 100 mg / dl or more. Here, the amount of scattered light is an arbitrary scale. As described above, the albumin concentration could be determined by measuring the scattered light amount of light propagating in urine. Thus, the scattering of light by relatively large particles such as albumin and blood is sufficiently larger than the scattering by urine glucose and other organic and inorganic substances. That is, the scattering of light propagating in urine is mainly caused by albumin or blood. Therefore, by irradiating urine with light and observing the amount of scattered light, albumin concentration and blood concentration can be determined. As described above, according to this example, the presence of large particles such as albumin and blood in urine can be inspected without using consumables such as test paper, and the practical effect is extremely high. large.

[Embodiment 4] This embodiment relates to a urine test method for simultaneously measuring the angle of rotation and the amount of scattered light of light propagating in urine. In particular, it is effective when glucose and albumin are present in urine to the extent that they cannot be ignored.
In the present embodiment, unlike the third embodiment, the amount of scattered light is observed in the form of a decrease in the amount of transmitted light. This embodiment is similar to the second embodiment.
The polarimeter shown in FIG. 3 described above was used as it was. The principle of the optical rotation angle measurement is the same as in the second embodiment. Hereinafter, a method of measuring the amount of transmitted light, that is, substantially the amount of scattered light will be described. When the output of the lock-in amplifier 10 is recorded while the rotation analyzer 7 is continuously rotated, the output change amount of the lock-in amplifier 10 per fixed rotation angle of the rotation analyzer 7 can be found. This output change amount corresponds to the transmitted light amount. By standardizing with the value when measuring pure water,
The amount of scattered light in urine can be grasped. By the above method,
With one measurement, the angle of rotation and the amount of scattered light can be simultaneously grasped, and the concentration of a light scattering substance such as albumin can be determined. That is, since the concentration range of albumin and the like necessary for Example 1 can be grasped, the glucose concentration can also be determined. As described above, according to the present embodiment, the glucose concentration and albumin concentration in urine can be simultaneously tested with a simple configuration and one measurement, and the practical effect is extremely large. As another method for measuring the amount of scattered light, for example, a rotary analyzer 7 is used.
Is temporarily fixed at a certain angle, and the output value of the lock-in amplifier 10 at this time can be grasped. Of course, the same effect can be obtained by directly detecting the scattered light from the side surface of the sample cell 6 as in the third embodiment. As explained above,
According to the present invention, it is possible to provide a urine test method that is easy to maintain without using consumables such as test paper.

The optical rotation angle increases as the wavelength of the measuring light becomes shorter until anomalous dispersion appears due to optical rotation dispersion. Therefore, it is more advantageous to use shorter wavelength light for more accurate measurement, but in the case of urine, it is generally 5%.
It is preferable to perform the process at a thickness of 00 nm or more. As shown in FIG. 8, which shows the spectral characteristics of normal urine, if the wavelength is shorter than 500 nm, the absorption mainly by urochrome (the yellow component contained in urine) increases, which may rather deteriorate the measurement accuracy. It is. Similarly, for absorption by chrome,
In measuring the amount of scattered light, the wavelength of the measurement light is preferably 500 nm or more.

[0027]

According to the present invention, it is possible to provide a urine test method that is easy to maintain without using consumables such as test paper.

[Brief description of the drawings]

FIG. 1 is a schematic diagram showing a configuration of a polarimeter used in Embodiment 1 of the present invention.

FIG. 2 is a characteristic diagram showing a relationship between an added glucose concentration and an optical rotation angle of an aqueous solution of glucose and urine in which glucose is dissolved.

FIG. 3 is a schematic diagram showing a configuration of a polarimeter used in Embodiment 2 of the present invention.

FIG. 4 is a characteristic diagram showing a relationship between the concentration of an aqueous solution of glucose and an aqueous solution of albumin and the angle of optical rotation with respect to light having a wavelength of 830 nm.

FIG. 5 is a characteristic diagram showing a relationship between the concentration of the aqueous solution and an optical rotation angle with respect to light having a wavelength of 589 nm.

FIG. 6 is a schematic diagram illustrating a configuration of a scattered light amount measuring device used in Embodiment 3 of the present invention.

FIG. 7 is a characteristic diagram showing the relationship between the concentration of an aqueous albumin solution and the amount of scattered light.

FIG. 8 is a characteristic diagram showing the relationship between the wavelength of incident light and the intensity of light transmitted through urine.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Sodium lamp 2 Band pass filter 3 Polarizer 4 Faraday cell 5 Faraday cell driver 6 Sample cell 7 Rotation analyzer 8 Rotation analyzer driver 9 Optical sensor 10 Lock-in amplifier 11 Computer 12 Semiconductor laser 13 Helium neon laser 14 Sample cell 15 light sensor

Continuation of the front page (56) References JP-A-5-203566 (JP, A) JP-A-61-145421 (JP, A) JP-A-7-294519 (JP, A) JP-A-6-510368 (JP, A) , A) (58) Field surveyed (Int. Cl. ⁷ , DB name) G01N 21/00-21/61 JOIS

Claims (4)

(57) [Claims] 1. A method for measuring the angle of rotation of urine in which the range of the angle of rotation expressed by an interfering substance other than a substance having an unknown concentration is known, and determining the glucose concentration C [kg / dl] by (A-A _h ) / (α × L) ≦ C ≦ (A- _Al ) / (α × L) where A: measured angle of rotation of urine [deg] A _h : angle of rotation generated by interfering optically rotating substance Maximum value [deg] _Al : Minimum value of the optical rotation angle generated by the interfering optically rotating substance [deg] α: Specific rotation of the optically rotating substance [deg / cm · dl / kg] L: Measurement optical path length [cm] A urine test method that determines the range. 2. The optical rotation angle of urine containing N kinds of optically rotating substances is measured by using light of at least N kinds of wavelengths, and the ratio of the optical rotation angle to the light of each wavelength and the ratio of each of the optically rotating substances is measured. determine the optical rotation angle range to be expressed by the interfering optical active substance by using the optical rotation, claim 1 urinalysis method according determining the concentration of the optical active substance in the urine. 3. The angle of rotation and the amount of scattered light of urine are measured simultaneously,
Optical rotation generated by the interfering optical rotation substance from the scattered light amount
Determines the angular range, the angle of rotation by optical rotation urinalysis method according to claim 1 or 2, wherein determining the concentration of a substance in said urine. 4. The urine test method according to claim 1, wherein the angle of rotation or the amount of scattered light of urine with respect to light having a wavelength of 500 nm or more is measured.