WO2023010613A1 - 一种用于定量检测液体生物样本中特定组分的弛豫核磁共振方法 - Google Patents
一种用于定量检测液体生物样本中特定组分的弛豫核磁共振方法 Download PDFInfo
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Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A90/00—Technologies having an indirect contribution to adaptation to climate change
- Y02A90/30—Assessment of water resources
Definitions
- the invention belongs to the technical field of analysis and detection of biochemical methods, and in particular relates to a relaxation nuclear magnetic resonance method for quantitatively detecting a specific component in a liquid biological sample, and a composition and a reaction solution for improving detection performance.
- Relaxation nuclear magnetic resonance technology has great application prospects in the fields of food science and biological detection because of its fast and easy operation.
- due to the low resolution of relaxation nuclear magnetic resonance it is difficult to distinguish multiple Therefore, the quantitative detection of small molecules in complex samples still has great limitations.
- the specificity, sensitivity, and Anti-interference has high requirements.
- One relaxation NMR detection method known in the art is the paramagnetic ion-mediated magnetic sensor, which amplifies the weak signal of the target component into a liquid through a target-induced Fe 2+ /Fe 3+ relaxation switch The change in relaxation of the sample as a whole.
- their sensitivity is still relatively low, which cannot meet the application requirements of trace component detection.
- the oxidation-reduction process of paramagnetic ions is easily interfered by levodopa, uric acid, vit-C, bilirubin, glutathione and other substances, resulting in false measurement results, seriously affecting the accuracy of the measurement sex.
- a solution proposed in the field is to pretreat biological samples by physical and chemical means to remove the interfering proteins as much as possible, but in practical applications, the samples Some small molecular compounds with redox properties cannot be simply eliminated, and redundant interfering components are easily introduced.
- a solution proposed in the art is to use the chelate of potassium thiocyanate and iron ions to enhance the contrast of the relaxation signal, but the formed complex is insoluble in water , It is easy to cause uneven sampling during detection and affect the measurement results. At the same time, potassium thiocyanate has certain toxicity and is harmful to the environment.
- the purpose of this invention is to further expand the application of relaxation nuclear magnetic resonance technology in the direction of biological detection, and propose a relaxation nuclear magnetic resonance method for quantitative detection of the content of specific components in liquid biological samples, and involves a series of methods using The composition and the prepared reaction solution for improving the sensitivity, specificity and anti-interference of relaxation detection, more specifically, convert the target component into the quantitative detection of the product hydrogen peroxide through the enzymatic reaction method, and pass the The composition enhances the 1 H-NMR relaxation signal difference caused by a trace amount of hydrogen peroxide, detects the change in the relaxation time of the reaction solution and compares the result with the standard tube to calculate the content of the target component.
- the trace specific component in the liquid biological sample that is, the target component, generates a hydrogen peroxide product related to its content
- the liquid biological sample is a sample derived from an organism and has a liquid texture
- the buffer in the specific enzymatic decomposition reaction is added with a composition to improve the detection performance, and the composition to improve the detection performance is composed of a specific type of enzyme, sorbitol, and ferrous salt;
- the liquid biological samples include whole blood, serum, urine, saliva, tissue fluid.
- the hydrogen peroxide products related to the generation of specific trace components refer to glucose and glucose oxidase, cholesterol ester and cholesterol esterase and cholesterol oxidase, triglyceride and lipase, glycerol kinase and phosphoglycerol oxidase. class combination.
- the relaxation NMR method refers to rapidly detecting information of a sample at least including transverse or longitudinal relaxation time based on 1 H-NMR time-domain signals.
- the standard tube sample of known concentration refers to a standard solution corresponding to the current target component, and the concentration of the standard is known.
- the specific type of enzyme is selected according to the current target component, including any one of glucose oxidase, cholesterol oxidase, and glycerol phosphate oxidase.
- Described ferrous salt comprises any one in ferrous chloride, ammonium ferrous sulfate.
- the final concentration of the specific type of enzyme is sufficient to specifically hydrolyze or oxidize the target component, the reaction process is accompanied by the product hydrogen peroxide, and the content of the final hydrogen peroxide product is proportional to the target component.
- the final concentration of ferrous ions produced by the ferrous salt is optimally prepared according to the content range of the target components, so that the ferrous ions in the final solution will not be completely oxidized.
- the present invention provides a method for rapidly detecting the content of specific components in liquid biological samples by nuclear magnetic resonance relaxation method.
- the composition of the present invention is The biochemical reaction between the components can improve the detection sensitivity and specificity of the relaxation nuclear magnetic resonance method for the trace components in the sample, and convert the content of the target component into the content of the product hydrogen peroxide, and then into the relaxation value measured by NMR.
- the change of Yu time greatly improves the contrast of detection difference.
- the NMR relaxation method adopted by the invention has high efficiency and low detection limit, and the related composition can improve the detection performance, and is more suitable for the detection of trace components in biological samples.
- Fig. 1 is the specific application of the reaction solution of the present invention and the composition therein in the quantitative detection of glucose in blood.
- Fig. 2 is the transverse relaxation curve and the corresponding relaxation time T 2 of the solution in the measurement tube of Example 1 of the present invention.
- Fig. 3 is the contrast of the result of measuring serum glucose content (embodiment 1) and the standard value (biochemical analyzer) by the method of the present invention, and its accuracy is better than commercially available common electronic blood glucose meters.
- the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.
- a relaxation nuclear magnetic resonance method for quantitatively detecting the content of a specific component in a liquid biological sample, a composition and a reaction solution for improving detection performance of the present invention are realized through the following technical scheme:
- a relaxation nuclear magnetic resonance method for quantitatively detecting the content of a specific component in a liquid biological sample the specific steps are:
- the liquid biological samples include whole blood, serum, urine, saliva, interstitial fluid and other samples derived from living organisms and having a liquid texture.
- Described specific component refers to the component that can specifically generate hydrogen peroxide product through one or several steps of enzymatic decomposition reaction, can be glucose (glucose oxidase), cholesterol ester (cholesteryl esterase, cholesterol esterase, cholesterol oxidase), triglycerides (lipase, glycerol kinase, phosphoglycerol oxidase), etc.
- the relaxation NMR method refers to rapid detection of transverse or longitudinal relaxation time of a sample based on 1 H-NMR of time-domain signals.
- the standard tube sample of known concentration refers to a standard solution corresponding to the current target component, and the concentration of the standard is known.
- the present invention provides the use of a composition in improving the detection performance of relaxation nuclear magnetic resonance.
- the improvement refers to the improvement of sensitivity, specificity, and/or anti-interference performance.
- the term "detection performance” mainly refers to sensitivity, specificity and anti-interference.
- the specific type of enzyme is selected according to the current target component, including but not limited to glucose oxidase, cholesterol oxidase, phosphoglycerol oxidase and the like.
- the ferrous salt can be dissolved in a buffer to generate ferrous ions, including but not limited to ferrous chloride, ammonium ferrous sulfate and the like.
- composition among the present invention are embodied by following biochemical reaction respectively:
- glucose ⁇ glucose oxidase ⁇ gluconic acid+H 2 O 2 said specific class of enzymes converts the detection of glucose into the detection of hydrogen peroxide with high specificity.
- triglyceride ⁇ lipase ⁇ glycerol + fatty acid, glycerol ⁇ glycerol kinase ⁇ glycerol-3-phosphate, glycerol-3-phosphate ⁇ glycerol phosphate oxidase ⁇ dihydroxyacetone phosphate + H 2 O 2 , said specific class of enzymes converts the detection of triglycerides to the detection of hydrogen peroxide with high specificity.
- Ferrous ions + H 2 O 2 ⁇ ferric ions the ferrous salt converts changes in hydrogen peroxide content into samples with high sensitivity through the ferromagnetic properties of ferrous ions/ferric ions The difference in relaxation time.
- the present invention provides a reaction solution for relaxation nuclear magnetic resonance detection, which is used for a series of high-sensitivity specific reactions with the target components in the detection sample, which contains the above-mentioned composition and a conventional buffer, Further samples are included.
- the final concentration of the specific type of enzyme is sufficient to cause specific hydrolysis or oxidation of the latter, the reaction process is accompanied by product hydrogen peroxide, and the content of the final hydrogen peroxide is proportional to the target component.
- the final concentration of ferrous ions produced by the ferrous salt is prepared according to the content range of the target components, so that the ferrous ions in the final solution will not be completely oxidized.
- the relaxation nuclear magnetic resonance method of the present invention quantitatively detects a specific component in a liquid biological sample.
- the trace nuclear magnetic signal of the specific component The difference is converted into a more significant relaxation signal difference caused by the oxidation of metal ions, which can detect the content of various components in liquid biological samples with high sensitivity, high specificity, and high anti-interference.
- the method can be applied to the rapid detection of multiple indicators.
- the glucose content in human serum samples was detected (detection range 3-20mmol/L).
- Embodiment result is as follows (italics are calculated data):
- the total cholesterol content in human serum samples was detected (detection range 2-10mmol/L).
- reaction solution suitable for quantitative detection of total cholesterol pH6.7 phosphate buffer 50mmol/L, cholesterol esterase ⁇ 0.4kU/L, cholesterol oxidase ⁇ 0.2kU/L, sodium cholate 0.1%, sorbic acid Alcohol 50mmol/L, ferrous ammonium sulfate 0.5mmol/L;
- the triglyceride content in human serum samples was detected (detection range 1-5 mmol/L).
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- Analytical Chemistry (AREA)
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Abstract
一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,具体过程包括步骤a):通过特异性的一步或几步酶分解反应使液体生物样本中微量目标组分生成与其含量相关的过氧化氢;步骤b):产物过氧化氢在组合物影响下高灵敏度地氧化亚铁离子并导致样本溶液测得的 1H-NMR弛豫时间降低;步骤c):与已知浓度的标准管样本比较并计算出被测样本中目标组分的含量。通过该方法,可以高灵敏度、高特异性、高抗干扰地检测液体生物样本中的多种组分含量,有助于进一步拓展核磁共振技术在生物检测方向的应用。
Description
本发明属于生物化学方法分析检测技术领域,具体涉及一种用于定量检测液体生物样本中特定组分的弛豫核磁共振方法及提升检测性能的组合物与反应液。
弛豫核磁共振技术因其快速、易操作的特点在食品科学、生物检测等领域具有极大的应用前景,但受限于弛豫核磁共振分辨率较低,时域上无法很好的区分多种组分的信号叠加,因此应用于复杂样本中的小分子定量检测仍具有很大局限。尤其是应用于生物样本中微量成分的检测时,易受到样本基质中不同水平的干扰物影响,导致检测误差或无法检出,因此对弛豫核磁共振方法在具体应用中的特异性、灵敏度、抗干扰性具有较高要求。
在本领域中已知的一种弛豫核磁共振检测方法是顺磁离子介导的磁传感器,通过目标诱导的Fe
2+/Fe
3+弛豫转换,将目标组分的微弱信号放大为液体样本整体的弛豫变化。但它们的灵敏度仍相对较低,无法满足微量组分检测的应用需求。同时在实际的临床应用中,顺磁离子的氧化还原过程易受到左旋多巴、尿酸、vit-C、胆红素、谷胱甘肽等物质的干扰造成假性测量结果,严重影响测定的准确性。在提升弛豫核磁共振方法的抗干扰性方面,本领域已提出的一种解决方法是通过物理化学手段对生物样本进行预处理,尽可能除去其中的干扰蛋白,但在实际应用中,样本中存在的一些具有氧化还原性质的小分子化合物并不能被简单除尽,还易引入多余的干扰组分。在提升弛豫核磁共振方法的灵敏性方面,本领域已提出的一种解决方法是利用硫***与铁离子的螯合物来增强弛豫信号对比度,但形成的络合物不溶于水,易造成检测时取样不均匀,影响测定结果。同时硫***具有一定毒性,对环境亦有危害。
在弛豫核磁共振检测领域,目前尚缺乏一种简单、安全、有效的方案,能够在针对液体生物样本的定量检测中同时提升检测性能的多个方面,包括但不限于针对葡萄糖、胆固醇、甘油三酯的微量检测灵敏度、抗干扰能力以及检测特异性等。
发明内容
技术问题:本发明的目的在于进一步拓展弛豫核磁共振技术在生物检测方向上的应用,提出一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,并涉及一系列用于提升弛豫检测灵敏度、特异性以及抗干扰性的组合物及配制形成的反应液,更具体地,是经由酶反应方法将目标组分转换为对产物过氧 化氢的定量检测,并通过所述组合物增强微量过氧化氢引起的
1H-NMR弛豫信号差异,检测反应液弛豫时间变化并与标准管结果比较计算出目标组分的含量。
技术方案:一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,具体步骤为:
a)通过特异性的酶分解反应使液体生物样本中的微量特定组分即目标组分生成与其含量相关的过氧化氢产物;所述的液体生物样本为来源于生物体且质地为液体的样本;所述的特异性的酶分解反应中的缓冲液里添加有提升检测性能的组合物,所述提升检测性能的组合物由特定种类酶、山梨醇、亚铁盐组成;
b)过氧化氢产物在组合物影响下高灵敏度地氧化亚铁离子并导致样本溶液测得的
1H-NMR弛豫时间降低;
c)与已知浓度的标准管样本检测结果比较并计算出被测样本中目标组分的含量。
所述液体生物样本包括全血、血清、尿液、唾液、组织液。
所述微量特定组分生成与其含量相关的过氧化氢产物指的是葡萄糖与葡萄糖氧化酶,胆固醇酯与胆固醇酯酶及胆固醇氧化酶,甘油三酯与脂肪酶、甘油激酶及磷酸甘油氧化酶这类组合。
所述弛豫核磁共振方法是指基于
1H-NMR时域信号快速检测样本至少包括横向或纵向弛豫时间在内的信息。
所述已知浓度的标准管样本是指对应当前目标组分的标准物溶液,标准物浓度已知。
所述特定种类酶根据当前目标组分选取,包括葡萄糖氧化酶、胆固醇氧化酶、磷酸甘油氧化酶中的任一种。
所述亚铁盐包括氯化亚铁、硫酸亚铁铵中的任一种。
所述特定种类酶的终浓度足以使目标组分发生特异性水解或氧化,反应过程中伴随有产物过氧化氢,且最终生成过氧化氢产物的含量与目标组分成比例关系。
所述亚铁盐产生的二价铁离子的终浓度根据目标组分的含量范围进行优化调配,使终溶液中二价铁离子不会被完全氧化。
本发明提供了一种通过核磁共振弛豫法检测快速检测液体生物样本中特定组分含量的方法,与现有检测液体样本的弛豫核磁共振技术相比,本发明中的组合物通过与目标组分间的生物化学反应可以提升弛豫核磁共振方法对样本中微 量组分的检测灵敏度和特异性,将目标组分的含量转化为产物过氧化氢的含量,继而转化为NMR测得的弛豫时间的变化,大大提高检测差异对比度。本发明采用的NMR弛豫法高效、检出限低,涉及的组合物可以提升检测性能,更适用于生物样本中微量组分的检测。
图1为本发明反应液及其中的组合物在针对血液中葡萄糖定量检测中的具体用途。
图2为本发明实施例1测定管中溶液的横向弛豫曲线及对应的弛豫时间T
2。
图3为本发明的方法测定血清葡萄糖含量(实施例1)的结果与标准值(生化分析仪)的对比,其准确度优于市售普通电子血糖仪。
以下结合说明书附图和具体实施例来进一步说明本发明,但实施例并不对本发明做任何形式的限定。除非特别说明,本发明采用的试剂、方法和设备均为本技术领域常规试剂、方法和设备。
除非特别说明,以下实施例所用试剂和材料均为市购。
本发明的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法及提升检测性能的组合物与反应液通过如下技术方案予以实现:
第一方面,一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,具体步骤为:
a)通过特异性的一步或几步酶分解反应使液体生物样本中的微量目标组分生成与其含量相关的过氧化氢;
b)产物过氧化氢在组合物影响下高灵敏度地氧化亚铁离子并导致样本溶液测得的
1H-NMR弛豫时间降低;
c)与已知浓度的标准管样本检测结果比较并计算出被测样本中目标组分的含量。
其中:
所述液体生物样本包括全血、血清、尿液、唾液、组织液等来源于生物体且质地为液体的样本。
所述特定组分是指可以经由一步或几步酶分解反应特异性地生成过氧化氢产物的组分,在具体的实施方案中可以是葡萄糖(葡萄糖氧化酶),胆固醇酯(胆固醇酯酶、胆固醇氧化酶),甘油三酯(脂肪酶、甘油激酶、磷酸甘油氧化酶) 等。
所述弛豫核磁共振方法是指基于时域信号的
1H-NMR快速检测样本横向或纵向弛豫时间。
所述已知浓度的标准管样本是指对应当前目标组分的标准物溶液,其标准物浓度已知。
第二方面,本发明提供一种组合物在提升弛豫核磁共振检测性能中的用途,所述组合物是指由包括特定种类酶、山梨醇、亚铁盐在内的生物化学试剂,有利地,所述提升是指灵敏度、特异性,和/或抗干扰性的性能的提升。
在本发明的定义中,术语“检测性能”主要是指灵敏度、特异性和抗干扰性。
其中:
所述特定种类酶根据当前目标组分选取,包括但不限于葡萄糖氧化酶、胆固醇氧化酶、磷酸甘油氧化酶等。
所述亚铁盐可以溶于缓冲液中产生二价铁离子,包括但不限于氯化亚铁、硫酸亚铁铵等。
本发明中组合物的用途分别通过如下生物化学反应予以体现:
在一个具体的实施方案中,胆固醇酯┄
胆固醇酯酶→胆固醇+脂肪酸,胆固醇┄
胆
固醇氧化酶→Δ
4-胆甾烯酮+H
2O
2,所述特定种类酶高特异性地将胆固醇酯和胆固醇的检测转化为过氧化氢的检测。
在一个具体的实施方案中,葡萄糖┄
葡萄糖氧化酶→葡萄糖酸+H
2O
2,所述特定种类酶高特异性地将葡萄糖的检测转化为过氧化氢的检测。
在一个具体的实施方案中,甘油三酯┄
脂肪酶→甘油+脂肪酸,甘油┄
甘油激酶→甘油-3-磷酸,甘油-3-磷酸┄
磷酸甘油氧化酶→磷酸二羟丙酮+H
2O
2,所述特定种类酶高特异性地将甘油三酯的检测转化为过氧化氢的检测。
二价铁离子+H
2O
2→三价铁离子,所述亚铁盐通过二价铁离子/三价铁离子不同的铁磁性质,高灵敏性地将过氧化氢含量的变化转化为样本弛豫时间的差异。
山梨醇+H
2O
2→过氧自由基,所述山梨醇与过氧化氢发生的链式反应提供的过氧自由基可以成倍氧化亚铁离子,极大提高反应液对过氧化氢的灵敏度。
第三方面,本发明提供一种弛豫核磁共振检测所用反应液,用于与检测样本中的目标组分发生一系列高灵敏性特异反应,其含有如上所述的组合物以及常规缓冲液,进一步包括样本。
其中:
所述特定种类酶的终浓度足以使后者发生特异性水解或氧化,反应过程中伴随有产物过氧化氢,且最终生成过氧化氢的含量与目标组分成比例关系。
所述亚铁盐产生的二价铁离子的终浓度根据目标组分的含量范围进行调配,使终溶液中二价铁离子不会被完全氧化。
本发明的弛豫核磁共振定量检测液体生物样本中特定组分的方法,与现有测定液体样本的弛豫核磁共振技术相比,通过引入本发明的组合物,将特定组分的微量核磁信号差异转化为金属离子氧化引起的更为显著的弛豫信号差异,可以高灵敏度、高特异性、高抗干扰性地检测液体生物样本中的多种组分含量。特别地,在复杂的临床检验情况下,通过调整不同的特定种类酶及对应的反应液,本方法可适用于多种指标的快速检测。
实施例1
检测人血清样本中的葡萄糖含量(检测范围3~20mmol/L)。
1.配制适用于葡萄糖定量检测的反应液:其中pH6.7磷酸盐缓冲液50mmol/L,葡萄糖氧化酶≥3ng/mL,山梨醇200mmol/L,氯化亚铁0.5mmol/L;
2.配制2只标准管样本液:其中葡萄糖浓度分别为3mmol/L以及20mmol/L,牛血清白蛋白0.2%;
3.取试管4只,按下表操作:
混匀后,37℃水浴孵育15min。
4.检测每只试管中溶液的横向弛豫时间T
2,计算全血葡萄糖(mmol/L)=3+17*(标准管1-测定管)/(标准管1-标准管2)。
实施例结果如下(斜体为计算数据):
标准管1 | 标准管2 | 测定管1 | 测定管2 | 测定管3 | 测定管4 | |
T 2/ms | 2059.75 | 218.43 | 1757.53 | 1293.44 | 1595.93 | 1866.5 |
Glucose/mM | 3 | 20 | 5.81 | 10.07 | 7.31 | 4.79 |
实施例2
检测人血清样本中的总胆固醇含量(检测范围2~10mmol/L)。
1.配制适用于总胆固醇定量检测的反应液:其中pH6.7磷酸盐缓冲液50mmol/L,胆固醇酯酶≥0.4kU/L,胆固醇氧化酶≥0.2kU/L,胆酸钠0.1%,山梨醇50mmol/L,硫酸亚铁铵0.5mmol/L;
2.配制2只标准管样本液:其中胆固醇标准物浓度分别为2mmol/L以及10mmol/L,牛血清白蛋白0.2%;
3.取试管4只,按下表操作:
混匀后,37℃水浴孵育15min。
4.检测每只试管中溶液的纵向弛豫时间T
1,计算血清总胆固醇(mmol/L)=2+8*(标准管1-测定管)/(标准管1-标准管2)。
实施例3
检测人血清样本中的甘油三酯含量(检测范围1~5mmol/L)。
1.配制适用于甘油三酯定量检测的反应液:其中pH6.7磷酸盐缓冲液50mmol/L,脂肪酶≥3U/mL,甘油激酶≥0.2U/mL,磷酸甘油氧化酶≥2.5U/mL,ATP≥0.15mmol/L,胆酸钠0.1%,山梨醇20mmol/L,硫酸亚铁铵0.5mmol/L;
2.配制2只标准管样本液:其中甘油三酯标准物浓度分别为1mmol/L以及5mmol/L,牛血清白蛋白0.2%;
3.取试管4只,按下表操作:
混匀后,37℃水浴孵育15min。
4.检测每只试管中溶液的纵向弛豫时间T
1,计算血清甘油三酯(mmol/L)=1+4*(标准管1-测定管)/(标准管1-标准管2)。
上述实施例为本发明针对血清中常见生化指标的定量检测的几种实施方式,但本发明的实施方式并不受上述实例的限制,其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化,均应为等效的置换方式,都包含在本发明的保护范围之内。
Claims (9)
- 一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,具体步骤为:a)通过特异性的酶分解反应使液体生物样本中的微量特定组分即目标组分生成与其含量相关的过氧化氢产物;所述的液体生物样本为来源于生物体且质地为液体的样本;所述的特异性的酶分解反应中的缓冲液里添加有提升检测性能的组合物,所述提升检测性能的组合物由特定种类酶、山梨醇、亚铁盐组成;b)过氧化氢产物在组合物影响下高灵敏度地氧化亚铁离子并导致样本溶液测得的 1H-NMR弛豫时间降低;c)与已知浓度的标准管样本检测结果比较并计算出被测样本中目标组分的含量。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述液体生物样本包括全血、血清、尿液、唾液、组织液。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述微量特定组分生成与其含量相关的过氧化氢产物指的是葡萄糖与葡萄糖氧化酶,胆固醇酯与胆固醇酯酶及胆固醇氧化酶,甘油三酯与脂肪酶、甘油激酶及磷酸甘油氧化酶这类组合。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述弛豫核磁共振方法是指基于 1H-NMR时域信号快速检测样本至少包括横向或纵向弛豫时间在内的信息。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述已知浓度的标准管样本是指对应当前目标组分的标准物溶液,标准物浓度已知。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述特定种类酶根据当前目标组分选取,包括葡萄糖氧化酶、胆固醇氧化酶、磷酸甘油氧化酶中的任一种。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述亚铁盐包括氯化亚铁、硫酸亚铁铵中的任一种。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述特定种类酶的终浓度足以使目标组分发生特异性水解或氧化,反应过程中伴随有产物过氧化氢,且最终生成过氧化氢产物的含量与目标组分成比例关系。
- 根据权利要求1所述的一种用于定量检测液体生物样本中特定组分含量的弛豫核磁共振方法,其特征在于,所述亚铁盐产生的二价铁离子的终浓度根据目标组分的含量范围进行优化调配,使终溶液中二价铁离子不会被完全氧化。
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WO2021089707A1 (en) * | 2019-11-07 | 2021-05-14 | Nanonord A/S | A method of and a system for determining protein concentration in a selected material by nuclear magnetic resonance relaxometry |
CN112067647A (zh) * | 2020-11-11 | 2020-12-11 | 东南大学 | 一种检测液体生物样本葡萄糖含量的弛豫核磁共振方法 |
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