Multi-mode determination hemoglobin eluent and determination method thereof

Abstract

The invention discloses a multi-mode hemoglobin determination eluent and a determination method thereof. The eluent comprises an eluent A, an eluent B, an eluent C, an eluent E, an eluent F and an eluent G which are used for determining hemoglobin, the eluent A is prepared from the following raw materials in percentage by weight: 0.08 to 0.12 percent of citric acid monohydrate, 0.40 to 0.50 percent of citric acid trialkali metal salt hydrate, 0.3 to 0.5 percent of perchloric acid alkali metal salt and 0.01 to 0.04 percent of sodium hydroxide. According to the method, on the premise that a stationary phase and a mobile phase of a chromatographic column do not need to be replaced, a mode of separating and measuring various hemoglobin by one eluent is realized, and the method can meet the measurement requirements of different types of hemoglobin, including conventional hemoglobin measurement, mutation hemoglobin measurement and thalassemia-related hemoglobin measurement.

Description

Technical Field

[0001] The present invention relates to the technical field of hemoglobin determination, and in particular to a multi-mode hemoglobin determination eluent and a determination method thereof. Background Art

[0002] Hemoglobin, especially glycosylated hemoglobin (HbA1c), which is a glycosylated protein obtained by binding hemoglobin to glucose, reflects the average blood sugar level over the past 1 to 2 months and is therefore widely used for the examination of lifestyle diseases including diabetes and metabolic syndrome and blood sugar management. Therefore, a method for easily and accurately measuring hemoglobin such as HbA1c is required.

[0003] In the prior art, there are high performance liquid chromatography (HPLC), immunoassay, enzyme method, electrophoresis method, etc. as methods for determining HbA1c. Among them, HPLC is often used in the field of clinical examination as a standard method for determining HbA1c. As a method for separating and determining hemoglobin using HPLC, it is common to use a cationic group chromatographic column as a stationary phase and a phosphate-based or succinic acid-based buffer as an eluent to analyze the hemoglobin in the sample, which can usually effectively separate and determine hemoglobin HbA1a, HbA1b, HbF, L-A1c, HbA1c, and HbA0. If hemoglobin other than the above is separated and determined, such as HbA2, HbE, HbD, HbS, and HbC, it is necessary to replace different chromatographic columns and eluents, which is very time-consuming and inconvenient in the use process and operation, which is not only inefficient, but also affects the accuracy and repeatability of the determination, such as Bio-Rad, Arkola, etc. Summary of the invention

[0004] In view of the problems existing in the prior art, the present invention provides a multi-mode hemoglobin determination eluent and a determination method thereof. The present invention realizes separation and determination modes of multiple hemoglobins with one eluent without replacing the chromatographic column stationary phase and mobile phase.

[0005] In order to achieve the above-mentioned object, the present invention provides a multi-mode hemoglobin determination eluent, wherein the eluent includes eluent A, eluent B, eluent C, eluent E, eluent F, and eluent G for determining hemoglobin;

[0006] The raw materials of the eluent A include 0.08wt%-0.12wt% of citric acid monohydrate, 0.40wt%-0.50wt% of tri-alkali metal salt of citric acid hydrate, 0.3wt%-0.5wt% of alkali metal perchlorate and 0.01wt%-0.04wt% of sodium hydroxide;

[0007] The raw materials of the eluent B include 0.01wt%-0.08wt% of citric acid monohydrate, 0.50wt%-0.60wt% of tri-alkali metal salt of citric acid hydrate, 1.0wt%-2.0wt% of alkali metal perchlorate and 0.02wt%-0.08wt% of sodium hydroxide;

[0008] The raw materials of the eluent C include 0.001wt%-0.01wt% of citric acid monohydrate, 0.50wt%-0.60wt% of tri-alkali metal salt of citric acid hydrate, 0.05wt%-0.15wt% of alkali metal perchlorate and 0.02wt%-0.08wt% of sodium hydroxide;

[0009] The eluent E is a mixture of eluent A and eluent C, with a weight ratio of 0.4-1.0;

[0010] The eluent F is a mixture of eluent B and eluent C, with a weight ratio of 0.1 to 0.8;

[0011] The eluent G is a mixture of the eluent C and the eluent A, and the weight ratio thereof is 0.25 to 0.5.

[0012] Preferably, the osmotic pressure of the eluent A is 100mOsm-200mOsm, the pH value is 5.0-6.0, and the conductivity is 8.2sm / cm-8.6sm / cm;

[0013] The osmotic pressure of the eluent B is 500mOsm-700mOsm, the pH value is 6.0-7.0, and the conductivity is 25sm / cm-35sm / cm;

[0014] The osmotic pressure of the eluent C is 10mOsm-200mOsm, the pH value is 6.0-7.0, and the conductivity is 3.0sm / cm to 6.0sm / cm.

[0015] Preferably, the hemoglobin is selected from one or more of HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC.

[0016] The present invention also provides a method for measuring hemoglobin eluate using the multi-mode measurement, comprising the following steps:

[0017] Step X1: Select the corresponding eluent or eluent combination and measurement mode according to the type of hemoglobin to be measured;

[0018] Step X2: inputting a filler into a high pressure liquid delivery pump, and using the high pressure liquid delivery pump to deliver a selected eluent or eluent combination into the chromatographic column;

[0019] Step X3: separating and measuring hemoglobin in a chromatographic column;

[0020] Preferably, the filler is a methacrylate copolymer, which is used to connect a high-pressure liquid delivery pump and an eluent and to separate and measure hemoglobin; the chromatographic column has a cation exchange group of one or more of sulfonyl, carboxyl and phosphate.

[0021] Preferably, the assay modes include rapid mode, rapid variation mode, variation assay mode, and thalassemia mode;

[0022] Among them, the rapid mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, and HbA0 hemoglobin;

[0023] The rapid mutation mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0 hemoglobin and mutant hemoglobin V-Win, where mutant hemoglobin V-Win is a general term for HbE, HbD, HbS and HbC;

[0024] The variation mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0 hemoglobin and HbE, HbD, HbS and HbC;

[0025] The thalassemia mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC.

[0026] Preferably, the steps in the fast mode are:

[0027] S1: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 2.0 mL / min and a delivery time of 25 s;

[0028] S2: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 2.0 mL / min and a delivery time of 10 s;

[0029] S3: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 20 s.

[0030] Preferably, the specific steps in the rapid mutation mode are:

[0031] S4: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 30 s;

[0032] S5: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 25 s;

[0033] S6: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 15 s;

[0034] S7: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 30 s.

[0035] Preferably, the specific steps in the mutation mode are:

[0036] S8: Use a high-pressure liquid delivery pump to deliver the eluent A to the separation chromatographic column: the flow rate is 1.8 mL / min, and the delivery time is 35 s;

[0037] S9: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 45 s;

[0038] S10: Use a high-pressure liquid delivery pump to deliver eluent A to the separation column at a flow rate of 1.8 mL / min and a delivery time of 10 s.

[0039] S11: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s;

[0040] S12: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s;

[0041] S13: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 30 s.

[0042] Preferably, the specific steps in the thalassemia model are:

[0043] S14: Use a high-pressure liquid delivery pump to deliver eluent A to the separation column at a flow rate of 1.8 mL / min and a delivery time of 35 s.

[0044] S15: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 25 s;

[0045] S16: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s;

[0046] S17: Use a high-pressure liquid delivery pump to deliver the eluent E to the separation chromatographic column, and continuously switch and mix the liquids through a high-frequency solenoid valve, with a flow rate of 1.8 mL / min and a time of 120 s;

[0047] S18: Use a high-pressure liquid delivery pump to deliver the eluent F to the separation chromatographic column, and continuously switch and mix the liquids through a high-frequency solenoid valve, with a flow rate of 1.8 mL / min and a time of 100 s;

[0048] S19: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 50 s.

[0049] The technical solution of the present invention has the following beneficial effects:

[0050] When the eluent of the present invention is used to separate and measure different hemoglobins, there is no need to replace the eluent. Only the liquid path timing control needs to be switched to achieve free combination, mixing and liquid delivery of the eluent, so as to achieve the same effect as replacing the reagent. It can meet the rapid detection requirements of the separation and measurement of HbA1a, HbA1b, HbF, L-A1c, HbA1c, and HbA0 hemoglobin, and can also meet the separation and measurement requirements of HbA2, HbE, HbD, HbS, and HbC.

[0051] The present invention has multi-mode applicability: by providing a variety of eluents and measurement modes, the present invention can adapt to different types of hemoglobin measurement needs, including conventional hemoglobin measurement, mutant hemoglobin measurement and thalassemia-related hemoglobin measurement.

[0052] The method of the present invention simplifies the operation process, does not need to replace the chromatographic column and the eluent, and can achieve efficient separation and determination of different hemoglobins by simply changing the combination and proportion of the eluent.

[0053] The present invention has high time efficiency: since the steps of replacing the chromatographic column and the eluent are reduced, the method of the present invention greatly shortens the measurement time and improves the work efficiency, and is particularly suitable for the needs of emergency and batch testing; the simplified operation process and reduced reagent consumption reduce the overall operating cost, making the method of the present invention more economical.

[0054] The eluent composition of the present invention is carefully designed to ensure the accuracy and repeatability of the measurement result and improve the reliability of the detection.

[0055] The eluent composition of the present invention is suitable for the determination of various hemoglobins, including but not limited to HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC, making it a powerful tool for hemoglobin research and clinical diagnosis. DETAILED DESCRIPTION

[0056] The present invention is further described below in conjunction with specific embodiments.

[0057] The present invention provides a multi-mode hemoglobin determination eluent, the eluent comprising eluent A, eluent B, eluent C, eluent E, eluent F, and eluent G for determining hemoglobin;

[0058] The raw materials of the eluent A include 0.08wt%-0.12wt% of citric acid monohydrate, 0.40wt%-0.50wt% of tri-alkali metal salt of citric acid hydrate, 0.3wt%-0.5wt% of alkali metal perchlorate and 0.01wt%-0.04wt% of sodium hydroxide;

[0059] The raw materials of the eluent B include 0.01wt%-0.08wt% of citric acid monohydrate, 0.50wt%-0.60wt% of tri-alkali metal salt of citric acid hydrate, 1.0wt%-2.0wt% of alkali metal perchlorate and 0.02wt%-0.08wt% of sodium hydroxide;

[0060] The raw materials of the eluent C include 0.001wt%-0.01wt% of citric acid monohydrate, 0.50wt%-0.60wt% of tri-alkali metal salt of citric acid hydrate, 0.05wt%-0.15wt% of alkali metal perchlorate and 0.02wt%-0.08wt% of sodium hydroxide;

[0061] The eluent E is a mixture of eluent A and eluent C, with a weight ratio of 0.4-1.0;

[0062] The eluent F is a mixture of eluent B and eluent C, with a weight ratio of 0.1 to 0.8;

[0063] The eluent G is a mixture of the eluent C and the eluent A, and the weight ratio thereof is 0.25 to 0.5.

[0064] Preferably, the osmotic pressure of the eluent A is 100mOsm-200mOsm, the pH value is 5.0-6.0, and the conductivity is 8.2sm / cm-8.6sm / cm;

[0065] Osmotic pressure (100mOsm-200mOsm): This range helps maintain the natural state of cells in the sample, reduces hemolysis, and ensures mild conditions during the elution process to avoid denaturation or degradation of hemoglobin; pH value (5.0-6.0): This pH value range provides a stable environment for hemoglobin, prevents it from changing under acidic conditions, and provides suitable pH conditions for chromatographic analysis; Conductivity (8.2sm / cm-8.6sm / cm): This conductivity range helps control the migration speed of ions in the eluent, optimizes separation efficiency, shortens analysis time, and ensures sufficient resolution.

[0066] The osmotic pressure of the eluent B is 500mOsm-700mOsm, the pH value is 6.0-7.0, and the conductivity is 25sm / cm-35sm / cm;

[0067] Osmotic pressure (500mOsm-700mOsm): Higher osmotic pressure helps achieve more efficient separation in specific chromatographic analyses, especially when analyzing larger molecules or hemoglobin variants that require higher elution strength. pH (6.0-7.0): The near-neutral pH range helps preserve the native structure of hemoglobin, avoiding denaturation that may occur under alkaline conditions, ensuring the accuracy of the assay.

[0068] Conductivity (25sm / cm-35sm / cm): Higher conductivity helps to speed up the migration of ions and improve separation efficiency, and is suitable for application scenarios that require rapid elution;

[0069] The osmotic pressure of the eluent C is 10mOsm-200mOsm, the pH value is 6.0-7.0, and the conductivity is 3.0sm / cm to 6.0sm / cm.

[0070] Conductivity (3.0sm / cm to 6.0sm / cm): Moderate conductivity range allows for faster separations while maintaining adequate resolution, ideal for applications that require a balance between analysis time and resolution.

[0071] Furthermore, the hemoglobin is selected from one or more of HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC; by measuring multiple hemoglobin variants, more comprehensive blood disease diagnostic information can be provided, such as diabetes control (through HbA1c), thalassemia (through HbA2), and other hereditary hemoglobinopathies (such as HbE, HbS). Early diagnosis: Certain hemoglobin variants such as HbA1c are indicators of long-term blood sugar control and help to detect diabetes-related complications early. Regular monitoring of the levels of specific hemoglobin variants is essential for evaluating the effectiveness of treatment and adjusting treatment plans.

[0072] The present invention also provides a method for measuring hemoglobin eluate using the multi-mode measurement, comprising the following steps:

[0073] Step X1: Select the corresponding eluent or eluent combination and measurement mode according to the type of hemoglobin to be measured;

[0074] Step X2: inputting a filler into a high pressure liquid delivery pump, and using the high pressure liquid delivery pump to deliver a selected eluent or eluent combination into the chromatographic column;

[0075] Step X3: separating and measuring hemoglobin in a chromatographic column;

[0076] Preferably, the filler is a methacrylate copolymer, which is used to connect a high-pressure liquid delivery pump and an eluent and to separate and measure hemoglobin; the chromatographic column has a cation exchange group of one or more of sulfonyl, carboxyl and phosphate.

[0077] Preferably, the assay modes include rapid mode, rapid variation mode, variation assay mode, and thalassemia mode;

[0078] Among them, the rapid mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, and HbA0 hemoglobin; it is easy to operate, takes a short time, and is suitable for rapid screening of a large number of samples. Application: Commonly used for routine physical examinations and rapid testing of outpatients.

[0079] The rapid mutation mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0 hemoglobin and mutant hemoglobin V-Win, where mutant hemoglobin V-Win is a general term for HbE, HbD, HbS and HbC;

[0080] The rapid mutation mode adds detection of mutant hemoglobin on the basis of the rapid mode, expands the detection range, and is suitable for the preliminary screening and diagnosis of hereditary hemoglobin diseases.

[0081] The variant mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0 hemoglobin and HbE, HbD, HbS and HbC; it provides more precise separation and measurement, and can detect more types of variant hemoglobin, which is suitable for the diagnosis and typing of patients suspected of hereditary hemoglobinopathy.

[0082] The thalassemia mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC. It can comprehensively cover the hemoglobin types related to thalassemia and provide detailed measurement results. It is suitable for the screening, diagnosis and treatment monitoring of thalassemia.

[0083] The present invention can adapt to different detection needs by providing multiple measurement modes, whether it is rapid screening or detailed diagnosis. The rapid mode and rapid variation mode greatly shorten the measurement time and improve the detection efficiency. The variation mode and thalassemia mode provide more precise separation and measurement, ensuring the accuracy of the results; the unified measurement process and automated operation reduce the complexity of the operation and lower the requirements for the operator's skills.

[0084] Preferably, the steps in the fast mode are:

[0085] S1: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 2.0 mL / min and a delivery time of 25 s;

[0086] S2: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 2.0 mL / min and a delivery time of 10 s;

[0087] S3: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 20 s.

[0088] Preferably, the specific steps in the rapid mutation mode are:

[0089] S4: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 30 s;

[0090] S5: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 25 s;

[0091] S6: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 15 s;

[0092] S7: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 30 s.

[0093] Preferably, the specific steps in the mutation mode are:

[0094] S8: Use a high-pressure liquid delivery pump to deliver the eluent A to the separation chromatographic column: the flow rate is 1.8 mL / min, and the delivery time is 35 s;

[0095] S9: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 45 s;

[0096] S10: Use a high-pressure liquid delivery pump to deliver eluent A to the separation column at a flow rate of 1.8 mL / min and a delivery time of 10 s.

[0097] S11: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s;

[0098] S12: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s;

[0099] S13: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 30 s.

[0100] Preferably, the specific steps in the thalassemia model are:

[0101] S14: Use a high-pressure liquid delivery pump to deliver eluent A to the separation column at a flow rate of 1.8 mL / min and a delivery time of 35 s.

[0102] S15: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 25 s;

[0103] S16: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s;

[0104] S17: Use a high-pressure liquid delivery pump to deliver the eluent E to the separation chromatographic column, and continuously switch and mix the liquids through a high-frequency solenoid valve, with a flow rate of 1.8 mL / min and a time of 120 s;

[0105] S18: Use a high-pressure liquid delivery pump to deliver the eluent F to the separation chromatographic column, and continuously switch and mix the liquids through a high-frequency solenoid valve, with a flow rate of 1.8 mL / min and a time of 100 s;

[0106] S19: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 50 s.

[0107] As can be seen from the above embodiments, when the eluent of the present invention is used to separate and measure different hemoglobin types, there is no need to replace the eluent. Only the liquid path timing control needs to be switched to achieve free combination, mixing and liquid delivery of the eluent, so as to achieve the same effect as replacing the reagent. It can meet the rapid detection requirements of the separation and measurement of HbA1a, HbA1b, HbF, L-A1c, HbA1c, and HbA0 hemoglobin, and can also meet the separation and measurement requirements of HbA2, HbE, HbD, HbS and HbC.

[0108] The present invention has multi-mode applicability: by providing a variety of eluents and measurement modes, the present invention can adapt to different types of hemoglobin measurement needs, including conventional hemoglobin measurement, mutant hemoglobin measurement and thalassemia-related hemoglobin measurement.

[0109] The method of the present invention simplifies the operation process, does not need to replace the chromatographic column and the eluent, and can achieve efficient separation and determination of different hemoglobins by simply changing the combination and proportion of the eluent.

[0110] The present invention has high time efficiency: since the steps of replacing the chromatographic column and the eluent are reduced, the method of the present invention greatly shortens the measurement time and improves the work efficiency, and is particularly suitable for the needs of emergency and batch testing; the simplified operation process and reduced reagent consumption reduce the overall operating cost, making the method of the present invention more economical.

[0111] The eluent composition of the present invention is carefully designed to ensure the accuracy and repeatability of the measurement result and improve the reliability of the detection.

[0112] The eluent composition of the present invention is suitable for the determination of various hemoglobins, including but not limited to HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC, making it a powerful tool for hemoglobin research and clinical diagnosis.

[0113] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent structural changes made by using the description and content of the present invention under the inventive concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A multi-mode hemoglobin eluate, characterized in that The eluents include eluent A, eluent B, eluent C, eluent E, eluent F, and eluent G for measuring hemoglobin; The raw materials of the eluent A include 0.08wt%-0.12wt% of citric acid monohydrate, 0.40wt%-0.50wt% of tri-alkali metal salt of citric acid hydrate, 0.3wt%-0.5wt% of alkali metal perchlorate and 0.01wt%-0.04wt% of sodium hydroxide; The raw materials of the eluent B include 0.01wt%-0.08wt% of citric acid monohydrate, 0.50wt%-0.60wt% of tri-alkali metal salt of citric acid hydrate, 1.0wt%-2.0wt% of alkali metal perchlorate and 0.02wt%-0.08wt% of sodium hydroxide; The raw materials of the eluent C include 0.001wt%-0.01wt% of citric acid monohydrate, 0.50wt%-0.60wt% of tri-alkali metal salt of citric acid hydrate, 0.05wt%-0.15wt% of alkali metal perchlorate and 0.02wt%-0.08wt% of sodium hydroxide; The eluent E is a mixture of eluent A and eluent C, with a weight ratio of 0.4-1.0; The eluent F is a mixture of eluent B and eluent C, with a weight ratio of 0.1 to 0.8; The eluent G is a mixture of the eluent C and the eluent A, and the weight ratio thereof is 0.25 to 0.

5.

2. The multimodal hemoglobin eluate according to claim 1, characterized in that: The osmotic pressure of the eluent A is 100mOsm-200mOsm, the pH value is 5.0-6.0, and the conductivity is 8.2sm / cm-8.6sm / cm; The osmotic pressure of the eluent B is 500mOsm-700mOsm, the pH value is 6.0-7.0, and the conductivity is 25sm / cm-35sm / cm; The osmotic pressure of the eluent C is 10mOsm-200mOsm, the pH value is 6.0-7.0, and the conductivity is 3.0sm / cm to 6.0sm / cm.

3. The multi-mode hemoglobin eluate according to claim 1, characterized in that: The hemoglobin is selected from one or more of HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC.

4. A method for measuring hemoglobin using the multimodal measurement hemoglobin eluate according to any one of claims 1 to 3, characterized in that: The steps include: Step X1: Select the corresponding eluent or eluent combination and measurement mode according to the type of hemoglobin to be measured; Step X2: inputting a filler into a high pressure liquid delivery pump, and using the high pressure liquid delivery pump to deliver a selected eluent or eluent combination into the chromatographic column; Step X3: Separating and measuring hemoglobin in a chromatographic column.

5. The method for multimodal determination of hemoglobin eluate according to claim 4, characterized in that: The filler is a methacrylate copolymer, which is used to connect a high-pressure liquid delivery pump and an eluent and to separate and measure hemoglobin; the cation exchange group of the chromatographic column is one or more of a sulfonic acid group, a carboxyl group and a phosphate group.

6. The method for multimodal determination of hemoglobin eluate according to claim 4, characterized in that: The measurement modes include rapid mode, rapid variation mode, variation measurement mode, and thalassemia mode; Among them, the rapid mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, and HbA0 hemoglobin; The rapid mutation mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0 hemoglobin and mutant hemoglobin V-Win, where mutant hemoglobin V-Win is a general term for HbE, HbD, HbS and HbC; The variation mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0 hemoglobin and HbE, HbD, HbS and HbC; The thalassemia mode is used to separate and measure HbA1a, HbA1b, HbF, L-A1c, HbA1c, HbA0, HbA2, HbE, HbD, HbS and HbC.

7. The method for multimodal determination of hemoglobin eluate according to claim 6, characterized in that: The steps in quick mode are: S1: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 2.0 mL / min and a delivery time of 25 s; S2: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 2.0 mL / min and a delivery time of 10 s; S3: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 20 s.

8. The method for multimodal determination of hemoglobin eluate according to claim 6, characterized in that: The specific steps in the rapid mutation mode are: S4: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 30 s; S5: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 25 s; S6: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 15 s; S7: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 30 s.

9. The method for multimodal determination of hemoglobin eluate according to claim 6, characterized in that: The specific steps in the mutation mode are: S8: Use a high-pressure liquid delivery pump to deliver the eluent A to the separation chromatographic column: the flow rate is 1.8 mL / min, and the delivery time is 35 s; S9: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 45 s; S10: Use a high-pressure liquid delivery pump to deliver eluent A to the separation column at a flow rate of 1.8 mL / min and a delivery time of 10 s. S11: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s; S12: Use a high-pressure liquid delivery pump to deliver eluent B to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s; S13: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 30 s.

10. The method for multimodal determination of hemoglobin eluate according to claim 6, characterized in that: Specific steps in the thalassemia model: S14: Use a high-pressure liquid delivery pump to deliver eluent A to the separation column at a flow rate of 1.8 mL / min and a delivery time of 35 s. S15: Use a high-pressure liquid delivery pump to deliver the eluent C to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 25 s; S16: Use a high-pressure liquid delivery pump to deliver eluent A to the separation chromatographic column at a flow rate of 1.8 mL / min and a delivery time of 10 s; S17: Use a high-pressure liquid delivery pump to deliver the eluent E to the separation chromatographic column, and continuously switch and mix the liquids through a high-frequency solenoid valve, with a flow rate of 1.8 mL / min and a time of 120 s; S18: Use a high-pressure liquid delivery pump to deliver the eluent F to the separation chromatographic column, and continuously switch and mix the liquids through a high-frequency solenoid valve, with a flow rate of 1.8 mL / min and a time of 100 s; S19: Use a high-pressure liquid delivery pump to deliver the eluent G to the separation chromatographic column, and use a high-frequency solenoid valve to continuously switch and mix the liquids at a flow rate of 1.8 mL / min for 50 s.