KR100776394B1 - Glycated hemoglobin cartridge and method for measuring glycated hemoglobin - Google Patents

Abstract

A glycated hemoglobin catridge is provided to separate the glycated hemoglobin contained in a blood sample rapidly and simply in one measurement space and be useful for allowing a glycated hemoglobin measuring device to quantitatively measure the total hemoglobin and the glycated hemoglobin in a sample. A glycated hemoglobin catridge(100) comprises a first storage container(110) containing hemolysis solution for hemolyzing blood; a second storage container(120) containing a sample solution including a glycated hemoglobin binding material-bead selectively reacting with the glycated hemoglobin; a top cover(130) including an inlet(131) to which a solution is injected and an insertion inducing portion(132); and a measurement cell(140) attached to the top cover, wherein the measurement cell includes a measuring space portion in which the glycated hemoglobin in the injected blood sample is separated by precipitating the glycated hemoglobin binding material-bead bound to the glycated hemoglobin generated by the reaction of the glycated hemoglobin in the hemolyzed blood sample injected from the first and second storage containers through the inlet with the glycated hemoglobin binding material-bead in the sample solution. A method for measuring glycated hemoglobin comprises the steps of: (a) putting a blood sample in the first storage container to hemolyze it; (b) injecting the hemolyzed blood into the measurement space portion of the measurement cell; (c) measuring the total hemoglobin in the hemolyzed blood sample; (d) injecting a sample solution including the glycated hemoglobin binding material-bead of the second storage container into the measurement space portion of the measurement cell; (e) isolating the glycated hemoglobin in the hemolyzed blood sample by precipitating the glycated hemoglobin bound to the glycated hemoglobin binding material-bead; (f) measuring the amount of the glycated hemoglobin isolated hemoglobin in the hemolyzed blood sample; and (g) comparing the amount of the total hemoglobin in the hemolyzed blood sample with the amount of the glycated hemoglobin isolated hemoglobin.

Description

Glycated Hemoglobin cartridge and method for measuring Glycated Hemoglobin}

1 is an example showing a glycated hemoglobin cartridge according to the present invention,

Figure 2 is an exploded perspective view of the glycated hemoglobin cartridge according to the present invention,

Figure 3 is an embodiment of use of the glycated hemoglobin cartridge of the present invention,

Figure 4 is a bottom view of the top cover of the glycated hemoglobin cartridge according to the present invention,

5 is a front view of a measuring cell of the glycated hemoglobin cartridge according to the present invention;

6 is a cross-sectional view of the glycated hemoglobin cartridge of the present invention;

Figure 7 is an exemplary view for explaining the process of separation of glycated hemoglobin and hemoglobin in the glycated hemoglobin cartridge of the present invention,

8 is a flowchart illustrating a method for measuring glycated hemoglobin present in hemolyzed blood samples using the glycated hemoglobin cartridge of the present invention;

9 is a graph showing results of the precipitation time by gravity according to the particle size of the glycated hemoglobin binding material-beads used in the present invention;

10 is a graph showing the results of the glycated hemoglobin reaction degree according to the particle size of the glycated hemoglobin binding material-beads used in the present invention;

11 is a graph showing the results of the separation of glycated hemoglobin according to the presence or absence of a stirring plate in the glycated hemoglobin cartridge according to the present invention.

<Brief Description of Drawings>

100: glycated hemoglobin cartridge

110: first storage container 120: second storage container

130: top cover

  131: inlet 132: insertion guide

  133: absorption pad attachment portion

140: measuring cell

  141: measuring space portion 142: sample space portion

  143: support groove 144: hemoglobin modified sample

150: stirring plate

The present invention relates to a technique for separating glycated hemoglobin in a blood sample, and more particularly, to a glycated hemoglobin cartridge and a method for separating glycated hemoglobin using the same.

Human red blood cells contain proteins for oxygen transport called hemoglobin. When blood sugar (glucose) rises in the blood, some of the glucose in the blood is bound to hemoglobin. This hemoglobin combined with glucose is called glycated hemoglobin (glycated Hemoglobin), hemoglobin A1c (HbA1c) is also called.

The normal life of red blood cells is about 120 days, and some of the red blood cells in our body are destroyed every day, and the amount of red blood cells similar to this destroyed amount is generated and maintained at a constant level. However, once glycated hemoglobin is created by combining with glucose, the red blood cells have a glycated hemoglobin until their life span is broken down.

Glucose testing is a test that checks how much sugar is in the blood every day, whereas glycated hemoglobin test reflects the average blood sugar level of eight weeks. In the case of blood glucose test, fasting blood glucose level should be fasted for at least 8 hours, and blood sugar level should be checked by blood collection 2 hours after eating, but glycated hemoglobin test should be done regardless of meal time. Has the advantage.

Since glycated hemoglobin test reflects a relatively long-term blood glucose level, it is used as an indicator to know whether diabetes has been well managed by treatment in recent months. In other words, the treatment goal of diabetes is to maintain normal blood sugar in order to prevent complications, and frequent blood glucose measurement is necessary to determine whether diabetes management is performed well.

However, measuring blood glucose frequently is cumbersome, so when blood glucose cannot be measured frequently, or when there is no need to measure it frequently, the glycated hemoglobin test can be used to determine the average blood sugar level over a long period of time. It is easy to see if this is going well. If the glycated hemoglobin level is high, it may be regarded as a condition that diabetes treatment is poorly performed, and the patient may manage diabetes by applying a stricter diet or increasing an insulin injection dose. .

By the way, the conventional glycated hemoglobin measurement was mostly measured in the clinical laboratory of the hospital, in this case, the sample pretreatment process, the volume of the equipment, the reagents and consumables were expensive.

In the case of US Pat. No. 5,162,237, a reactant blocked by a cover flows out of the reaction channel inside the bio cartridge, and the sample is measured at one corner, and then the sample is reacted at the other corner. Therefore, the measurement must take place sequentially at timed intervals, and the person taking the measurement must intervene in the measurement phase by pulling the cover in order of the experiment so that the reactant flows into the test sample.

In the case of US Pat. No. 6,300,142, the test sample is reacted to the first reactant through the first inlet, and subsequently to the second reactant through the second inlet to measure two or more analytes present in the sample. A device for measuring two analytes is disclosed. Even in this case, the measurements must take place sequentially, at timed intervals, and the measuring person must intervene in the measurement step to respond by sequentially injecting test samples-the measurement is complicated because the beads combined with glycated hemoglobin must be filtered out once. It takes a long time.

The present invention has been proposed in the background as described above, and an object of the present invention is to provide a glycated hemoglobin cartridge capable of quickly and simply separating glycated hemoglobin contained in a blood sample in one measurement space.

Furthermore, an additional object of the present invention is to provide a glycated hemoglobin cartridge which can quantitatively measure the amount of total hemoglobin and glycated hemoglobin contained in a blood sample in a glycated hemoglobin measurement apparatus.

Another object of the present invention is to provide a method for separating and measuring glycated hemoglobin contained in a blood sample using a glycated hemoglobin cartridge having one measurement space.

A glycated hemoglobin cartridge according to an aspect of the present invention for achieving the above object comprises a first storage container containing a hemolytic blood hemolytically, a sample comprising a glycated hemoglobin binding material-bead that reacts selectively with glycated hemoglobin A second reservoir containing the solution, a top cover including an inlet into which the solution is injected, and a measuring cell attached to the top cover, wherein the measuring cell is in the blood sample injected from the first and second reservoirs through the inlet; Glycated hemoglobin binding material in the glycated hemoglobin and the sample solution-glycated hemoglobin binding material combined with the glycated hemoglobin produced by the reaction between the beads-characterized in that it comprises a measurement space for separating the glycated hemoglobin in the blood sample precipitated and injected do. According to this aspect, the glycated hemoglobin cartridge according to the present invention can quickly and simply separate the glycated hemoglobin contained in the blood sample in one measurement space.

In the method of measuring glycated hemoglobin according to another aspect of the present invention, a blood sample is collected from a human body into a first storage container to hemolyse blood, and the hemolyzed blood of the first storage container is injected into a measurement space of a measurement cell. Step, measuring the total amount of hemoglobin in the hemolyzed blood sample injected into the measurement space of the measurement cell by using the glycated hemoglobin measurement device, measuring the sample solution containing the glycated hemoglobin binding material-bead of the second storage container Injecting into the measuring space of the cell, the glycated hemoglobin binding material in the blood injected into the measuring space of the measuring cell and the glycated hemoglobin binding material in the sample solution-the glycated hemoglobin binding material combined with the glycated hemoglobin produced by the beads Isolating the glycated hemoglobin in the hemolyzed blood sample, the glycated hemoglobin in the hemolyzed blood sample using a glycated hemoglobin measuring device The amount of hemoglobin in the hemolyzed blood sample was calculated by comparing the amount of hemoglobin in the hemolyzed blood sample with the step of measuring the amount of hemoglobin separated and the amount of hemoglobin in the hemolyzed blood sample using the glycated hemoglobin measurement device. Characterized in that it comprises a step.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily understand and reproduce the present invention.

1 is a perspective view of a glycated hemoglobin cartridge according to the present invention, Figure 2 is an exploded view of the glycated hemoglobin cartridge according to the present invention, Figure 3 is an embodiment of use of the glycated hemoglobin cartridge of the present invention.

As shown in FIG. 1, the glycated hemoglobin cartridge 100 of the present invention includes a first storage container 110, a second storage container 120, a top cover 130, and a measurement cell 140.

The first storage container 110 contains a hemolytic fluid for hemolysis of blood, and the second storage container 120 contains a sample solution containing glycated hemoglobin binding material-beads that selectively react with glycated hemoglobin. The first storage container 110 and the second storage container 120 may be provided separately from the top cover 130 and the measurement cell 140, or may be packaged and integrally provided.

Here, the hemolysin is a buffer solution containing a surfactant, such as 20 mM Hepes buffer solution (N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic Acid HEPES; pH 8.1). Hemoglobin and glycated hemoglobin are present in the hemolyzed blood sample. The glycated hemoglobin binding material may be embodied as, for example, boronic acid (BA), concanavalin A (Lectin), or an antibody that can specifically bind to glycated hemoglobin. Beads are polymeric polysaccharide supports such as agarose or cellulose, latex beads or glass such as polystyrene, polymethylmethacrylate, polyvinyltolune, or polyvinyltolune. It can be implemented as a glass bead.

The top cover 130 includes an inlet 131 through which the solution is injected, and an insertion guide part 132 that guides the glycated hemoglobin cartridge to be accurately inserted into the glycated hemoglobin measuring device. The top cover 130 is inclined toward the inlet 131 in which the inlet 131 is formed, and is made of a material having low light transmittance. The top cover 130 is implemented to be attached to the measurement cell 140, and further, the top cover 130 and the measurement cell 140 may be rotatably coupled.

The measurement cell 140 receives the blood sample and the sample solution contained in the first and second storage containers 110 and 120 through the inlet 131 of the top cover 130. The measurement cell 140 is preferably made of a material having high light transmittance in order to increase the accuracy of the colorimetric measurement performed in the glycated hemoglobin measurement device (reference numeral 50 of FIG. 3).

As shown in FIG. 2, the measurement cell 140 is formed in the glycated hemoglobin in the blood sample and the sample solution injected from the first and second storage containers 110 and 120 through the inlet 131 of the top cover 130. A glycated hemoglobin binding material-a glycated hemoglobin binding material combined with a glycated hemoglobin generated according to the reaction between the beads-includes a measurement space 141 in which the glycated hemoglobin in the injected blood is separated. Here, the glycated hemoglobin binding material combined with glycated hemoglobin-beads may be precipitated by gravity or electromagnetic force acting on the beads. Reference numeral 142 denotes a sample space part which prevents the blood sample after the measurement from flowing out of the measurement cell.

As shown in FIG. 2, the glycated hemoglobin cartridge 100 of the present invention may further include a stirring plate 150 that moves according to external force generated by the glycated hemoglobin measurement device 50, for example, centrifugal force and electromagnetic force. In this case, the measurement cell 140 further includes a support groove 143 supporting the stirring plate 150 disposed in the measurement space 141. When the external force generated in the glycated hemoglobin measuring device 50 is an electromagnetic force, the stirring plate 150 should be implemented with an electric conductor such as silver, copper, aluminum, iron, or an alloy mainly composed of copper. The stirring plate 150 allows the reaction between the glycated hemoglobin of the hemolyzed blood and the glycated hemoglobin binding material-bead of the sample solution in the measurement space 141 to be sufficient in a short time.

In one embodiment, as shown in Figure 2, the measurement cell 140 may be applied to the hemoglobin modified sample 144 on the inner wall of the measurement space 141. Hemoglobin modified sample 144 is potassium ferricyanide (K ₃ Fe (CN) ₆ ) and potassium cyanide (KCN) or potassium ferricyanide (K ₃ Fe (CN) ₆ ) and lithium thioncyanide (LiSCN) or anionic surfactant ( It can be implemented with sodium laurylsulfate (anionic surfactant) and Triton X-100 (TritonX-100) which is a nonionic surfactant. Hemoglobin present in various types of hemolyzed blood reacts with potassium cyanide (K ₃ Fe (CN) ₆ ) and potassium cyanide (KCN) applied to the inner wall of the measurement space 141 to cyanmet-hemoglobin (cyanmet-hemoglobin, HbCN), sodium laurylsulfate-hemoglobin (SLS-hemoglobin), azidemet-hemoglobin. Cyanmet-hemoglobin (HbCN), sodium laurylsulfate-hemoglobin (SLS-hemoglobin), which is formed by reacting the hemoglobin modified sample 144 with hemoglobin in hemolyzed blood Azidemet-hemoglobin can be measured by colorimetric measurement of the amount of total hemoglobin in the hemolyzed blood.

As shown in Fig. 3, the glycated hemoglobin cartridge 100 of the present invention is used in the glycated hemoglobin measurement device 50. The glycated hemoglobin measuring device 50 used in the present invention measures the amount of total hemoglobin in the blood sample and the amount of hemoglobin from which the glycated hemoglobin is separated according to a known colorimetric method. In other words, hemoglobin specifically absorbs light signals of a certain frequency. According to the properties of hemoglobin, the glycated hemoglobin measuring device 50 used in the present invention measures the amount of hemoglobin using a light emitting device such as a photodiode and a light receiving device. The glycated hemoglobin measurement device 50 is preferably implemented to move the stirring plate (reference numeral 150 of FIG. 2) by generating an external force, for example, centrifugal force and electromagnetic force. The glycated hemoglobin cartridge 100 of the present invention is mounted in accordance with the insertion groove 51 of the glycated hemoglobin measurement device 50 corresponding to the insertion guide unit 132.

Figure 4 is a bottom view of the top cover of the glycated hemoglobin cartridge according to the present invention. As shown, the top cover 130 is an inlet 131 into which the solution is injected and the insertion guide portion 132 to guide the glycated hemoglobin cartridge to be accurately inserted into the glycated hemoglobin measuring device on its outer surface and the absorption pad on the inner surface thereof. An absorbent pad attaching part 133 to be detached and attached is formed. In this case, the absorption pad absorbs the blood sample after the measurement and prevents the blood sample from flowing out of the measurement cell.

5 is a front view of the measuring cell of the glycated hemoglobin cartridge according to the present invention. The glycated hemoglobin cartridge of the present invention can be used for the purpose of correcting the zero point of the glycated hemoglobin measurement device and for measuring the glycated hemoglobin. In one example, the measurement cell 140 is used to correct the zero point of the glycated hemoglobin measurement device when the recognition device 200 is attached. The measuring cell 140 is used for measuring glycated hemoglobin when the recognition device 200 is detached. The measurement cell 140 is formed on the outer bottom surface of the recognition device mounting portion is detached, attached to the recognition device 200 to allow the glycated hemoglobin measuring device to recognize the type and installation of the glycated hemoglobin cartridge. Here, the recognition device mounting portion may be implemented as a mounting groove or an adhesive member.

6 is a cross-sectional view of the glycated hemoglobin cartridge according to the present invention. As shown, the glycated hemoglobin cartridge 600 according to the present invention is implemented such that the solution is injected into the measurement space 641 of the measurement cell 640 through the inlet 631 of the top cover 630. In addition, the glycated hemoglobin cartridge 600 is agitation plate 650 agitated the blood and the sample solution injected into the measurement space 641, the glycated hemoglobin binding material of the blood glycated hemoglobin and the sample solution in the measurement space 641 -Ensure that the reaction between beads is sufficient in a short time.

Figure 7 is an exemplary view for explaining the process of separation of glycated hemoglobin and hemoglobin in the glycated hemoglobin cartridge of the present invention.

First, the blood sample is put into the first storage container 81 in which the blood collected from the human body is contained, and the blood sample is hemolyzed. Hemoglobin and glycated hemoglobin are present in the hemolyzed blood sample. Thereafter, as shown in FIG. 7I, the blood sample hemolyzed in the first storage container 81 is injected into the measurement space part 70 of the measurement cell. Here, reference numeral 71 is a stirring plate, 72 is a glycated hemoglobin, 73 is a hemoglobin.

Subsequently, as shown in FIG. 7, the sample solution containing the glycated hemoglobin binding material-bead contained in the second storage container 82 is injected into the measurement space part 70 of the measurement cell. Here, reference numeral 74 denotes a glycated hemoglobin binding material-bead. After injecting the hemolyzed blood sample and the sample solution containing the glycated hemoglobin binding material-bead into the measurement space part 70, the stirring plate 72 is moved to stir the hemolyzed blood sample and the glycated hemoglobin binding material-bead. While agitation occurs, the measurement space 70 generates a reaction between glycated hemoglobin-binding material-beads and glycated hemoglobin-binding material-beads 75 in which glycated hemoglobin is bound.

Subsequently, as shown in III of FIG. 7, when a certain time elapses, the agitation plate 72 stops the movement and the glycated hemoglobin binding material-bead combined with the glycated hemoglobin present in the measurement space 70 is moved by gravity. Allow to settle. As such, glycated hemoglobin binding material combined with glycated hemoglobin-beads precipitate by gravity to separate hemoglobin and hemoglobin in hemolyzed blood. The present invention improves the degree of reaction between the glycated hemoglobin and glycated hemoglobin binding material-beads contained in the hemolyzed blood in the measurement space 70 and the precipitation time due to the gravity of the glycated hemoglobin-binding material bound to the glycated hemoglobin-beads. It is necessary to appropriately select the size of the particle size of the glycated hemoglobin binding agent-beads.

8 is a flowchart illustrating a method for measuring glycated hemoglobin present in hemolyzed blood samples using the glycated hemoglobin cartridge of the present invention.

First, the blood sample taken from the human body is put into a first storage container containing hemolyzed blood to hemolyse the blood (S801). The hemolyzed blood of the first storage container is injected into the measurement space of the measurement cell (S802). Using the glycated hemoglobin measuring device to measure the total amount of hemoglobin in the hemolyzed blood sample injected into the measurement space of the measurement cell (S803).

In the glycated hemoglobin cartridge of the present invention, a hemoglobin modified sample may be applied to the inner wall of the measurement space of the measurement cell. In other words, the amount of total hemoglobin in the blood sample may be measured by colorimetric measurement of hemolyzed blood, but hemoglobin present in various types may be modified by a hemoglobin modified sample applied to the inner wall of the measurement space of the measurement cell. Hemoglobin can be measured by colorimetry. Herein, the hemoglobin modified sample is potassium ferricyanide (K ₃ Fe (CN) ₆ ) and potassium cyanide (KCN) or potassium ferricyanide (K ₃ Fe (CN) ₆ ), lithium cyanide (LiSCN), anionic surfactant (anionic) Sodiumlaurylsulfate (surfactant) and triton X-100 (TritonX-100), which is a nonionic surfactant.

Thereafter, the sample solution containing the glycated hemoglobin binding material-bead of the second storage container is injected into the measurement space of the measurement cell (S804). Glycated hemoglobin in the blood injected into the measurement space of the measuring cell and glycated hemoglobin binding material in the sample solution-glycated hemoglobin binding material combined with the glycated hemoglobin produced in response to the beads-glycated hemoglobin in the hemolyzed blood sample by precipitation It is separated (S805). Here, the glycated hemoglobin binding material associated with glycated hemoglobin-beads is precipitated by gravity acting on the beads. In step S805 of separating the glycated hemoglobin, a stirring plate provided in the measurement space is formed so that the reaction between the glycated hemoglobin of the blood and the glycated hemoglobin binding material of the sample solution-beads is sufficiently performed in the measurement space of the measurement cell. Move during the reaction time to agitate the blood and sample solution injected into the measurement space.

A glycated hemoglobin measurement device is used to measure the amount of hemoglobin in which the glycated hemoglobin is separated in the hemolyzed blood sample (S806). The glycated hemoglobin measuring device used in the present invention measures the amount of total hemoglobin in the blood sample according to the colorimetric measuring method already well known. The amount of glycated hemoglobin in the hemolyzed blood sample is calculated by comparing the amount of total hemoglobin in the hemolyzed blood sample with the amount of the hemoglobin from which the glycated hemoglobin is separated (S807).

Figure 9 is a graph showing the results of the precipitation time by gravity by the particle size of the glycated hemoglobin binding material-beads used in the present invention, Figure 10 is the degree of glycated hemoglobin response by particle size of the glycated hemoglobin binding material-beads used in the present invention A graph of the results of the implementation.

Applicants have a carboxyl molecule for experiments and have Sepharose (hereinafter referred to as CM sepharose) having various particle size sizes, 3-aminophenylboronic acid and EDC. (1-Ethyl-3-carbodiimide hydrochloride) or EDAC (1-Ethyl-3-dimethylaminopropyl hydrochloride), buffered solution, washing solution, hemolyzed solution, and 6% and 9% glycated hemoglobin standard samples were used.

In more detail, 6 w / v% CM sepharose is washed with 100 mM scalpel buffer (4-Morpholineethanesulfonic acid) pH 4.7 to prepare 6 w / v%. 800 mM EDC and 66 mM 3-aminophenyl boronic acid were also prepared in a mass buffer solution, respectively. Prepare a sample solution containing glycated hemoglobin binding material-beads by reacting the prepared CM sepharose with EDC and 3-aminophenyl boronic acid. After the reaction, wash with a mass buffer solution and react with 4M sodium acetate and 50mM NaOH to remove carboxyl functional groups remaining without 3-aminophenyl boronic acid in CM sepharose.

Synthesized glycated hemoglobin binding agent--beads (N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic Acid, HEPES) as much as 20 mM to meet conditions that can react with actual glycated hemoglobin in blood Wash thoroughly and store.

20 mM Hepes buffer solution (N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic Acid, HEPES) at pH8.1, 0.1% sodium azide, 0.05% TritonX-100 After adding the hemolysis to prepare the hemolyzed 900 ul put in a storage container.

20 mM Hepes buffer solution (N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic Acid, HEPES) at pH8.1, 0.1% sodium azide, 0.05% TritonX-100 To prepare the sample solution containing the glycated hemoglobin-binding material-bead to add 1.3 w / v% to each 600 ul prepared in a tube.

Prepare 6% and 9% glycated hemoglobin standard samples, respectively, identified with G7 as reference equipment. Store the glycated hemoglobin standard solution in cryogenic freezer below -70 ℃.

6% and 9% glycated hemoglobin standard samples were hemolyzed in hemolyzed solution and placed in storage containers respectively. Thereafter, a sample solution containing glycated hemoglobin binding material-beads is introduced into each reservoir, followed by stirring. After 6% and 9% of the glycated hemoglobin standard solution and the sample solution are reacted for a predetermined time, the time taken for the glycated hemoglobin conjugated-beads to be precipitated is measured. As shown in Figure 9, when the particle size of the CM sepharose is 50㎛ or less, the time to settle by gravity increases rapidly.

Meanwhile, 6% and 9% glycated hemoglobin standard samples were measured in the hemolyzed blood, and then the amount of hemoglobin and the amount of hemoglobin from which the glycated hemoglobin was separated were measured by colorimetry, and the glycated hemoglobin was separated from the total hemoglobin level. The amount of hemoglobin is calculated by subtracting the amount of hemoglobin. As shown in Fig. 10, it can be seen that the degree of reaction between the glycated hemoglobin and the sample solution decreases when the particle size of the glycated hemoglobin binding material-bead is 500 µm or more.

Therefore, the glycated hemoglobin binding material-particle size size is preferably selected in consideration of the precipitation time of the glycated hemoglobin binding material and the glycated hemoglobin combined with the glycated hemoglobin after the reaction, in the present invention, glycated hemoglobin binding Substance-The particle size of beads is 50-500 탆.

Figure 11 is a graph of the results of the separation of glycated hemoglobin according to the presence or absence of a stirring plate in the glycated hemoglobin cartridge according to the present invention.

Applicants measured glycated hemoglobin in various glycated hemoglobin standard samples in a glycated hemoglobin cartridge with a stir plate and a glycated hemoglobin cartridge without a stir plate. In Fig. 11, diamond (◆) is a result of the separation of glycated hemoglobin using a glycated hemoglobin cartridge with a stirring plate, and rectangle (■) is a result of the separation of glycated hemoglobin using a glycated hemoglobin cartridge without a stirring plate. . As shown, it can be seen that the glycated hemoglobin cartridge with the stirring plate is better separated than the glycated hemoglobin cartridge without the stirring plate.

As described above, the glycated hemoglobin cartridge according to the present invention has a useful effect of quickly and simply separating and measuring the glycated hemoglobin contained in the hemolyzed blood sample in one measurement space.

In addition, the glycated hemoglobin cartridge according to the present invention has a useful effect of allowing the glycated hemoglobin measuring device to quantitatively measure the total hemoglobin and glycated hemoglobin contained in the sample.

Although the present invention has been described with reference to the accompanying drawings, it will be apparent to those skilled in the art that many various obvious modifications are possible without departing from the scope of the invention from this description. Therefore, it should be interpreted by the claims described to include many such variations.

Claims (17)

In the glycated hemoglobin cartridge used in the glycated hemoglobin measuring device, The glycated hemoglobin cartridge is: A first reservoir container containing hemolytic fluid for hemolysis of blood; A second storage container containing a sample solution containing glycated hemoglobin binding material that selectively reacts with glycated hemoglobin-beads; A top cover including an inlet through which a solution is injected; And A measurement cell attached to the top cover, The measurement cell, The glycated hemoglobin binding material in the hemolyzed hemoglobin sample injected from the first and second storage vessels through the inlet and the glycated hemoglobin binding material in the sample solution-the glycated hemoglobin binding material bound to the glycated hemoglobin produced in response to the beads- Glycosylated hemoglobin cartridge characterized in that it comprises a measurement space that is precipitated to separate the glycated hemoglobin in the injected blood sample. The method of claim 1, Glycosylated hemoglobin binding material coupled to the glycated hemoglobin-bead, Glycated hemoglobin cartridge, characterized in that precipitated by gravity acting on the beads. The method of claim 1, wherein the glycated hemoglobin cartridge is: Further comprising a stirring plate moving in accordance with the external force generated in the glycated hemoglobin measurement device, The measurement cell, The glycated hemoglobin cartridge further comprises a support groove for supporting the stirring plate disposed in the measurement space on an upper surface thereof. The method of claim 2, The glycated hemoglobin cartridge, characterized in that the external force generated in the glycated hemoglobin measuring device is an electromagnetic force. The method of claim 1, The measurement cell, The glycated hemoglobin cartridge further comprises a recognition device mounting portion to which the hemoglobin measuring device recognizes the type and installation of the glycated hemoglobin cartridge on its outer bottom surface. The method of claim 1, The top cover, A glycated hemoglobin cartridge further comprising an absorption pad on an inner surface thereof to absorb a solution flowing out of the measurement cell. The method of claim 1, The top cover, The glycated hemoglobin cartridge further comprises an insertion inducing unit for inducing the glycated hemoglobin cartridge to be accurately inserted into the glycated hemoglobin measuring device. The method of claim 1, The top cover, Glycosylated hemoglobin cartridge, characterized in that the outer surface formed with the inlet is inclined toward the inlet, made of a material having a low light transmittance. The method of claim 1, The glycated hemoglobin binding material-bead, A glycated hemoglobin cartridge, characterized in that the average particle size of the beads is about 50 to 500㎛. The method of claim 1, The glycated hemoglobin binding material-bead, The bead is at least one of a polysaccharide beads, latex (latex) beads or glass (glass) beads A glycated hemoglobin cartridge characterized in that. The method of claim 1, The glycated hemoglobin binding material-bead, The glycated hemoglobin binding material is at least one of boronic acid (boronic acid), concanavalin A (concanavalin A), the antibody (antibody) A glycated hemoglobin cartridge characterized in that. The method of claim 1, On the inner wall of the measuring space part of the measuring cell, Hemoglobin hemoglobin cartridge characterized in that the hemoglobin modified sample reacted with hemoglobin in the hemolyzed blood sample injected from the first reservoir through the inlet. The method of claim 12, The hemoglobin modified sample, At least potassium ferricyanide (K ₃ Fe (CN) ₆ ) and potassium cyanide (KCN), potassium ferricyanide (K ₃ Fe (CN) ₆ ) and lithium thioncyanide (LiSCN), sodium laurylsulfate and triton X Any one of -100 (TritonX-100) A glycated hemoglobin cartridge characterized in that. In the method for measuring glycated hemoglobin present in hemolyzed blood samples using a glycated hemoglobin cartridge used in the glycated hemoglobin measuring device, The glycated hemoglobin cartridge is a first storage container containing a hemolytic blood for hemolysis, a second storage container containing a sample solution containing a glycated hemoglobin binding material-beads that selectively react with glycated hemoglobin, and an inlet for solution injection It includes a top cover and a measuring cell attached to the top cover, Glycosylated hemoglobin in the sample solution and glycosylated hemoglobin in the sample solution-the glycosylated hemoglobin bound to the glycosylated hemoglobin produced in response to the bead-from the first and second reservoirs through the inlet; A measurement space portion in which beads are precipitated to separate glycated hemoglobin in the injected blood sample, The glycated hemoglobin measurement method is: Placing a blood sample taken from a human body into the first storage container to hemolyze blood; Injecting the hemolyzed blood of the first reservoir into a measurement space of the measurement cell; Measuring the amount of total hemoglobin in the hemolyzed blood sample injected into the measurement space of the measurement cell by using the glycated hemoglobin measurement device; Injecting a sample solution containing the glycated hemoglobin binding material-bead of the second reservoir into a measurement space of the measurement cell; The glycated hemoglobin binding material formed by the reaction between the glycated hemoglobin binding material in the sample solution and the glycated hemoglobin binding material-bead injected into the measurement space of the measurement cell was precipitated and precipitated in the hemolyzed blood sample. Separating glycated hemoglobin; Measuring the amount of hemoglobin in which glycated hemoglobin is separated in the hemolyzed blood sample using the glycated hemoglobin measuring device; And Calculating the amount of glycated hemoglobin in the hemolyzed blood sample by comparing the amount of total hemoglobin in the hemolyzed blood sample with the amount of the hemoglobin from which the glycated hemoglobin was separated using the apparatus for measuring glycated hemoglobin; Glycated hemoglobin measurement method comprising a. The method of claim 14, A method for measuring glycated hemoglobin, characterized in that the glycated hemoglobin binding material combined with the glycated hemoglobin-bead is precipitated by gravity acting on the beads. The method of claim 14, The glycated hemoglobin cartridge further comprises a stirring plate moving according to the external force generated in the glycated hemoglobin measuring device, The measuring cell further comprises a support groove for supporting the stirring plate in the measuring space portion on the upper surface thereof, The step of separating the glycated hemoglobin is: Stirring the blood and the sample solution injected into the measurement space of the measurement cell by moving the stirring plate of the measurement cell for a predetermined reaction time; Method for measuring glycated hemoglobin, characterized in that it further comprises. The method of claim 14, On the inner wall of the measuring space part of the measuring cell, A hemoglobin modified sample reacting with hemoglobin in the hemolyzed blood sample injected from the first reservoir through the inlet is applied, Measuring the amount of total hemoglobin in the hemolyzed blood sample includes: Hemoglobin hemoglobin, characterized in that for measuring the amount of hemoglobin in the hemolyzed blood using a hemoglobin hemoglobin measuring device produced by reacting the hemoglobin modified hemoglobin in the hemoglobin modified sample How to measure.

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