CN106908411A - A kind of urea in serum nitrogen content near infrared ray method - Google Patents
A kind of urea in serum nitrogen content near infrared ray method Download PDFInfo
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Abstract
本发明公开了一种血清中尿素氮含量近红外光谱测定方法，步骤一、配制若干个含有不同尿素氮含量的血清标定液；步骤二、用近红外光谱仪对每个血清标定液进行光谱采集；步骤三、根据血清标定液的近红外光谱，建立鉴别模型；步骤四、用近红外光谱仪对待检测的血清样品进行光谱采集，将血清样品的光谱与鉴别模型对比，获得血清样品中的尿素氮含量。本发明具有检测时间短、检测效率高、检测准确性高等优点。The invention discloses a method for measuring urea nitrogen content in serum by near-infrared spectroscopy. Step 1: Prepare several serum calibration solutions containing different urea nitrogen contents; Step 2: Use a near-infrared spectrometer to collect the spectrum of each serum calibration solution; Step 3, according to the near-infrared spectrum of the serum calibration solution, establish an identification model; step 4, use a near-infrared spectrometer to collect the spectrum of the serum sample to be detected, compare the spectrum of the serum sample with the identification model, and obtain the urea nitrogen content in the serum sample . The invention has the advantages of short detection time, high detection efficiency, high detection accuracy and the like.
Description
技术领域technical field
本发明属于血液检测领域，涉及一种血清中尿素氮含量的测定方法，尤其涉及一种血清中尿素氮含量近红外光谱测定方法。The invention belongs to the field of blood detection, and relates to a method for measuring urea nitrogen content in serum, in particular to a method for measuring urea nitrogen content in serum by near-infrared spectroscopy.
背景技术Background technique
尿素是人体蛋白质代谢的主要终末产物，氨基酸脱氨基产生NH3和C02，两者在肝脏中合成尿素。通常肾脏为排泄尿素的主要器官，尿素从肾小球滤过后在各段小管均可重吸收。各种肾实质性病变，如肾小球肾炎、间质性肾炎、急慢性肾功能衰竭等均可使血尿素氮增高。如能排除肾外因素，尿素氮可作为尿毒症诊断指标之一。Urea is the main end product of human protein metabolism, amino acid deamination produces NH 3 and C0 2 , and the two synthesize urea in the liver. Usually, the kidney is the main organ for excreting urea, and urea can be reabsorbed in various tubules after being filtered from the glomerulus. Various renal parenchymal diseases, such as glomerulonephritis, interstitial nephritis, acute and chronic renal failure, etc., can increase blood urea nitrogen. If extrarenal factors can be excluded, blood urea nitrogen can be used as one of the diagnostic indicators of uremia.
血清(或血浆)中的尿素氮，在尿素氮试剂的酸性环境中与二乙酰-肟(DAM)共沸后，可缩合成一红色化合物，称为fearon反应。其颜色的深浅与血清(或血浆)中尿素氮的含量成正比，与同样处理的尿素氮标准液比色，即可测算出血清(或血浆)中尿素氮的含量。由于此方法干扰性大，目前临床实验室多采用尿素酶偶联法，用尿素酶分解尿素产生氨，氨在谷氨酸脱氢酶的作用下使NADH氧化为NAD+时，通过340nm吸光度的降低值可计算出尿素氮含量。但是，尿素酶偶联法测试时间较长，步骤繁琐，检测效率慢，且大量的人为操作易造成检测结果不准确。The urea nitrogen in serum (or plasma) can be condensed into a red compound after being azeotroped with diacetyl-oxime (DAM) in the acidic environment of urea nitrogen reagent, which is called fearon reaction. The depth of its color is directly proportional to the content of urea nitrogen in serum (or plasma), and the content of urea nitrogen in serum (or plasma) can be measured by colorimetric comparison with the same treated urea nitrogen standard solution. Due to the high interference of this method, the current clinical laboratory mostly adopts the urease coupling method, which uses urease to decompose urea to produce ammonia . The reduced value can be used to calculate the urea nitrogen content. However, the urease coupling method takes a long time to test, the steps are cumbersome, the detection efficiency is slow, and a large number of human operations are likely to cause inaccurate detection results.
发明内容Contents of the invention
本发明提供一种血清中尿素氮含量近红外光谱测定方法，以克服现有技术的缺陷。The invention provides a method for measuring blood urea nitrogen content by near-infrared spectroscopy to overcome the defects of the prior art.
为实现上述目的，本发明提供一种血清中尿素氮含量近红外光谱测定方法，步骤一、配制若干个含有不同尿素氮含量的血清标定液；步骤二、用近红外光谱仪对每个血清标定液进行光谱采集；步骤三、根据血清标定液的近红外光谱，建立鉴别模型；步骤四、用近红外光谱仪对待检测的血清样品进行光谱采集，将血清样品的光谱与鉴别模型对比，获得血清样品中的尿素氮含量。In order to achieve the above object, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, step 1, preparing several serum calibration solutions containing different urea nitrogen contents; step 2, using a near-infrared spectrometer for each serum calibration solution Carry out spectral collection; Step 3, establish a discrimination model according to the near-infrared spectrum of the serum calibration solution; Step 4, use a near-infrared spectrometer to collect the spectrum of the serum sample to be detected, compare the spectrum of the serum sample with the discrimination model, and obtain the serum sample urea nitrogen content.
其中，步骤一中，血清标定液是指已知其尿素氮含量的血清样品。且血清标定液的数量需满足建立模型的要求。优选的，血清标定液的数量至少为30个。若干个血清标定液的尿素氮含量的范围为2-7mmol/L。Wherein, in step 1, the serum calibration solution refers to a serum sample whose urea nitrogen content is known. And the quantity of serum standard solution needs to meet the requirements of model establishment. Preferably, the number of serum standard solutions is at least 30. The urea nitrogen content of several serum calibration solutions ranged from 2-7mmol/L.
步骤三中，鉴别模型是指血清标定液的光谱与血清标定液中尿素氮含量之间的对应关系。In step 3, the identification model refers to the corresponding relationship between the spectrum of the serum calibration solution and the content of urea nitrogen in the serum calibration solution.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，步骤一是采用尿素酶偶联法测定血清中的尿素氮含量，并配制出不同尿素氮含量的血清标定液；尿素酶偶联法是指采用尿素酶、谷氨酸脱氢酶、α-酮戊二酸、NADH与血清混合，根据NADH与血清反应前后的变化，即依据NADH与血清反应前后的结果，得到血清中尿素氮的含量。Further, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which may also have the following characteristics: wherein, the first step is to use urease coupling method to measure urea nitrogen content in serum, and prepare different urea nitrogen content of serum calibration solution; urease coupling method refers to the use of urease, glutamate dehydrogenase, α-ketoglutaric acid, NADH and serum mixed, according to the changes before and after the reaction between NADH and serum, that is, according to the NADH and serum The results before and after the reaction were obtained to obtain the content of blood urea nitrogen in the serum.
其中，上述血清可以为采集而来的多份血清，用以配制血清标定液。Wherein, the above-mentioned serum can be multiple serums collected to prepare the serum calibration solution.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，尿素酶偶联法的具体步骤为：步骤a、试剂与血清混合，并于36～38℃温浴3-6min；试剂包括尿素酶、谷氨酸脱氢酶、α-酮戊二酸和NADH；步骤b、在340nm波长处测量试剂和血清的混合液中NADH的吸光度下降速率，并与相同处理的尿素氮标准液比较，计算获得样品中尿素氮的含量。Further, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which may also have the following characteristics: wherein, the specific steps of the urease coupling method are as follows: step a, mixing reagents with serum, and 36-38 Incubate at ℃ for 3-6min; reagents include urease, glutamate dehydrogenase, α-ketoglutarate and NADH; step b, measure the absorbance decrease rate of NADH in the mixed solution of reagent and serum at 340nm wavelength, and compare with The content of urea nitrogen in the sample is calculated by comparing the urea nitrogen standard solution with the same treatment.
其中，步骤b中，“与相同处理的尿素氮标准液比较”是指：配置尿素氮标准液，加入试剂，混合均匀并温浴，其中，尿素氮标准液与血清等量，试剂的加入量与其加入至血清中的量相等，温浴温度和时间也与样品的处理条件相同。然后，在340nm波长处测量试剂和尿素氮标准液的混合液中NADH的吸光度下降速率，由于尿素氮标准液的浓度已知，通过计算可知血清中尿素氮的含量。并且，尿素氮标准液可以与血清同时处理，以使检测结果更加准确并节省检测时间。Wherein, in step b, "comparing with the urea nitrogen standard solution of the same treatment" refers to: configuring the urea nitrogen standard solution, adding reagents, mixing evenly and warming the bath, wherein, the urea nitrogen standard solution is equal to the serum, and the addition amount of the reagent is the same as that of the serum. The amount added to the serum was equal, and the incubation temperature and time were also the same as the treatment conditions of the samples. Then, measure the absorbance decrease rate of NADH in the mixed solution of the reagent and the urea nitrogen standard solution at a wavelength of 340nm. Since the concentration of the urea nitrogen standard solution is known, the content of urea nitrogen in the serum can be known by calculation. Moreover, the blood urea nitrogen standard solution can be processed at the same time as the serum to make the test results more accurate and save test time.
尿素氮标准液的配制方法为：称取尿素，溶于无氨去离子水中，并用无氨去离子水定容。尿素氮标准液的浓度为6～8mmol/L。The preparation method of urea nitrogen standard solution is as follows: weigh urea, dissolve it in ammonia-free deionized water, and use ammonia-free deionized water to make up to volume. The concentration of urea nitrogen standard solution is 6-8mmol/L.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，试剂还包括磷酸盐缓冲液；磷酸盐缓冲液的pH指为7.7±0.2；优选的，磷酸盐缓冲液为TRIS-盐酸缓冲剂。Further, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which may also have the following characteristics: wherein, the reagent also includes a phosphate buffer; the pH of the phosphate buffer is 7.7±0.2; preferably, Phosphate buffer is TRIS-HCl buffer.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，试剂还包括防腐剂；优选的，防腐剂为ProcIin-300。Further, the present invention provides a method for measuring blood urea nitrogen content in serum by near-infrared spectroscopy, which may also have such a feature: wherein, the reagent also includes a preservative; preferably, the preservative is Proclin-300.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，试剂由磷酸盐缓冲液30～70nmol/L、尿素酶4000～6000U/L、谷氨酸脱氢酶8000～10000U/L、α-酮戊二酸140～180nmol/L、NADH2.0～4.0nmol/L、防腐剂150～250μl/L组成；试剂与血清的体积比为1:70～80。Further, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which may also have the following characteristics: wherein, the reagents are 30-70 nmol/L of phosphate buffer, 4000-6000 U/L of urease, glutamic acid Dehydrogenase 8000～10000U/L, α-ketoglutarate 140～180nmol/L, NADH2.0～4.0nmol/L, preservative 150～250μl/L; the volume ratio of reagent to serum is 1:70～ 80.
其中，试剂由上述各组分混合制成。Wherein, the reagent is prepared by mixing the above-mentioned components.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，步骤二和步骤四中，使用近红外光谱仪检测血清标定液和血清样品时，还采用配合近红外光谱仪的液体检测装置；液体检测装置包括通光孔、半透半反镜片和样品通道探测器；通光孔、半透半反镜片和样品通道探测器设于同一直线上，形成样品测试光路。Further, the present invention provides a method for measuring blood urea nitrogen content in serum by near-infrared spectroscopy, which may also have the following characteristics: wherein, in step 2 and step 4, when using a near-infrared spectrometer to detect the serum calibration solution and serum samples, a combined method is also used. A liquid detection device for a near-infrared spectrometer; the liquid detection device includes a light hole, a semi-transparent mirror and a sample channel detector; the light hole, the semi-transparent mirror and the sample channel detector are arranged on the same straight line to form a sample test light path.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，液体检测装置还包括空气通道探测器；空气通道探测器设于半透半反镜片的一侧，与样品测试光路垂直的方向，用于接收半透半反镜片的反射光。Further, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which may also have the following characteristics: wherein, the liquid detection device also includes an air passage detector; Side, the direction perpendicular to the light path of the sample test, is used to receive the reflected light from the half-transparent and half-reflective mirror.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，液体检测装置还包括样品试管；样品试管放置在半透半反镜片和样品通道探测器之间，位于样品测试光路上；进行近红外光谱采集时，血清标定液或待检测的血清样品均放入样品试管中。Further, the present invention provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which may also have such characteristics: wherein, the liquid detection device also includes a sample test tube; the sample test tube is placed between the semi-transparent and semi-reflective lens and the sample channel detector The space is located on the optical path of the sample test; when collecting near-infrared spectra, the serum calibration solution or the serum sample to be tested are placed in the sample tube.
检测时，近红外光线通过通光孔，射至半透半反镜片上，一部分近红外光线由半透半反镜片反射进入空气通道探测器；另一部分近红外光线透过半透半反镜片照射样品试管，透过样品试管及其内部的测试液体(血清标定液或血清样品)射入样品通道探测器，光线在透过测试液体时发生吸收现象。During detection, the near-infrared light passes through the light hole and hits the semi-transparent and semi-reflective lens, part of the near-infrared light is reflected by the semi-transparent and semi-reflective lens and enters the air channel detector; the other part of the near-infrared light passes through the semi-transparent and semi-reflective lens to irradiate the sample The test tube is injected into the sample channel detector through the sample test tube and the test liquid (serum calibration solution or serum sample) inside, and light absorption occurs when the light passes through the test liquid.
进一步，本发明提供一种血清中尿素氮含量近红外光谱测定方法，还可以具有这样的特征：其中，样品试管呈长方体，测试光线与样品试管的管壁垂直。Further, the present invention provides a near-infrared spectroscopic method for measuring blood urea nitrogen content, which may also have the following characteristics: wherein, the sample test tube is in the shape of a cuboid, and the test light is perpendicular to the tube wall of the sample test tube.
本发明的有益效果在于：本发明提供一种血清中尿素氮含量近红外光谱测定方法，采集若干个不同尿素氮含量血清的近红外光谱，建立尿素氮含量与光谱相对应的鉴别模型，当需要检测其他血清样品的尿素氮含量时，仅需采集该血清样品的近红外光谱，并将其与上述鉴别模型对比，即可获得该血清样品中的尿素氮含量。与传统的标准检测方法相比，首先，本方法的待检测血清样品无需预处理，不使用任何化学试剂，无环境污染、成本低；其次，检测所需的血清样品量少，且每份样品的检测时间约为近红外光谱的采集时间，检测时间短、效率高；此外，本方法避免了人为操作的误差，提高检测准确率。The beneficial effect of the present invention is: the present invention provides a kind of near-infrared spectrum determination method of urea nitrogen content in serum, collects the near-infrared spectrum of several different urea nitrogen content serums, establishes the identification model corresponding to urea nitrogen content and spectrum, when needs When detecting the urea nitrogen content of other serum samples, it is only necessary to collect the near-infrared spectrum of the serum sample and compare it with the above identification model to obtain the urea nitrogen content of the serum sample. Compared with the traditional standard detection method, firstly, the serum sample to be tested in this method does not require pretreatment, does not use any chemical reagents, has no environmental pollution, and has low cost; secondly, the amount of serum sample required for detection is small, and each sample The detection time is about the acquisition time of the near-infrared spectrum, and the detection time is short and the efficiency is high; in addition, this method avoids the error of human operation and improves the detection accuracy.
具体实施方式detailed description
实施例一Embodiment one
本实施例提供一种血清中尿素氮含量近红外光谱测定方法：The present embodiment provides a method for measuring blood urea nitrogen content by near-infrared spectroscopy:
步骤一、采用尿素酶偶联法测定血清中的尿素氮含量，并配制出若干个不同尿素氮含量的血清标定液。上述血清可以为采集而来的多份血清，用以配制血清标定液。Step 1: Use the urease coupling method to measure the urea nitrogen content in the serum, and prepare several serum calibration solutions with different urea nitrogen contents. The above-mentioned serum can be collected multiple serums to prepare serum calibration solution.
其中，血清标定液的数量需满足建立模型的要求。优选的，血清标定液的数量至少为30个。Among them, the quantity of serum calibration solution needs to meet the requirements of model establishment. Preferably, the number of serum standard solutions is at least 30.
若干个血清标定液的尿素氮含量的范围为2-7mmol/L。The urea nitrogen content of several serum calibration solutions ranged from 2-7mmol/L.
尿素酶偶联法的具体步骤为：步骤a、试剂与血清混合，并于36℃温浴6min。其中，试剂由磷酸盐缓冲液30nmol/L、尿素酶4000U/L、谷氨酸脱氢酶8000U/L、α-酮戊二酸140nmol/L、NADH2.0nmol/L、防腐剂150μl/L组成。其中，磷酸盐缓冲液为TRIS-盐酸缓冲剂，防腐剂为ProcIin-300。血清与试剂的体积比为1:70。步骤b、在340nm波长处测量试剂和血清的混合液中NADH的吸光度下降速率，并与相同处理的尿素氮标准液比较，计算获得样品中尿素氮的含量。The specific steps of the urease coupling method are as follows: step a, mixing the reagent and serum, and incubating at 36° C. for 6 minutes. Among them, the reagent is composed of phosphate buffer 30nmol/L, urease 4000U/L, glutamate dehydrogenase 8000U/L, α-ketoglutarate 140nmol/L, NADH 2.0nmol/L, preservative 150μl/L . Wherein, the phosphate buffer is TRIS-hydrochloric acid buffer, and the preservative is Proclin-300. The volume ratio of serum to reagent is 1:70. Step b. Measure the decrease rate of absorbance of NADH in the mixture of reagent and serum at a wavelength of 340nm, and compare it with the same treated urea nitrogen standard solution to calculate the content of urea nitrogen in the sample.
其中，尿素氮标准液的浓度为6mmol/L。在尿素氮标准液中加入试剂，混合均匀并温浴，其中，尿素氮标准液与血清等量，试剂的加入量与其加入至血清中的量相等，温浴温度和时间也与样品的处理条件相同。然后，在340nm波长处测量试剂和尿素氮标准液的混合液中NADH的吸光度下降速率，由于尿素氮标准液的浓度已知，通过计算可知血清中尿素氮的含量。Among them, the concentration of urea nitrogen standard solution is 6mmol/L. Add reagents to urea nitrogen standard solution, mix well and warm bath, wherein, the urea nitrogen standard solution is equal to the amount of serum, the amount of reagent added is equal to the amount added to the serum, and the temperature and time of the warm bath are also the same as the processing conditions of the sample. Then, measure the absorbance decrease rate of NADH in the mixed solution of the reagent and the urea nitrogen standard solution at a wavelength of 340nm. Since the concentration of the urea nitrogen standard solution is known, the content of urea nitrogen in the serum can be known by calculation.
步骤二、用近红外光谱仪对每个血清标定液进行光谱采集。Step 2, use a near-infrared spectrometer to collect the spectrum of each serum calibration solution.
步骤三、根据血清标定液的近红外光谱，建立鉴别模型。该鉴别模型是指血清标定液的光谱与血清标定液中尿素氮含量之间的对应关系。Step 3: Establish an identification model based on the near-infrared spectrum of the serum calibration solution. The identification model refers to the corresponding relationship between the spectrum of the serum calibration solution and the content of urea nitrogen in the serum calibration solution.
步骤四、用近红外光谱仪对待检测的血清样品进行光谱采集，将血清样品的光谱与鉴别模型对比，获得血清样品中的尿素氮含量。Step 4: Use a near-infrared spectrometer to collect the spectrum of the serum sample to be tested, compare the spectrum of the serum sample with the identification model, and obtain the urea nitrogen content in the serum sample.
其中，步骤二和步骤四中，使用近红外光谱仪检测血清标定液和血清样品时，还采用配合近红外光谱仪的液体检测装置。该液体检测装置包括通光孔、半透半反镜片、样品试管、样品通道探测器和空气通道探测器。Wherein, in step 2 and step 4, when using the near-infrared spectrometer to detect the serum calibration solution and serum samples, a liquid detection device that cooperates with the near-infrared spectrometer is also used. The liquid detection device comprises a light hole, a semi-transparent and semi-reflective lens, a sample test tube, a sample channel detector and an air channel detector.
通光孔、半透半反镜片和样品通道探测器设于同一直线上，形成样品测试光路。空气通道探测器设于半透半反镜片的一侧，与样品测试光路垂直的方向，用于接收半透半反镜片的反射光。样品试管放置在半透半反镜片和样品通道探测器之间，位于样品测试光路上，进行近红外光谱采集时，血清标定液或待检测的血清样品均放入样品试管中。样品试管呈长方体，测试光线与样品试管的管壁垂直。The light hole, the semi-transparent and semi-reflective mirror and the sample passage detector are arranged on the same straight line to form a sample testing optical path. The air channel detector is arranged on one side of the semi-transparent and semi-reflective lens, in the direction perpendicular to the light path of the sample test, and is used to receive the reflected light of the semi-transparent and semi-reflective lens. The sample test tube is placed between the semi-transparent and semi-reflective lens and the sample channel detector, and is located on the sample test optical path. When performing near-infrared spectrum collection, the serum calibration solution or the serum sample to be tested are placed in the sample test tube. The sample test tube is a cuboid, and the test light is perpendicular to the wall of the sample test tube.
检测时，近红外光线通过通光孔，射至半透半反镜片上，一部分近红外光线由半透半反镜片反射进入空气通道探测器；另一部分近红外光线透过半透半反镜片照射样品试管，透过样品试管及其内部的测试液体(血清标定液或血清样品)射入样品通道探测器，光线在透过测试液体时发生吸收现象。During detection, the near-infrared light passes through the light hole and hits the semi-transparent and semi-reflective lens, part of the near-infrared light is reflected by the semi-transparent and semi-reflective lens and enters the air channel detector; the other part of the near-infrared light passes through the semi-transparent and semi-reflective lens to irradiate the sample The test tube is injected into the sample channel detector through the sample test tube and the test liquid (serum calibration solution or serum sample) inside, and light absorption occurs when the light passes through the test liquid.
实施例二Embodiment two
本实施例提供一种血清中尿素氮含量近红外光谱测定方法，与实施例一的方法基本相同，区别仅在于：步骤一中的尿素酶偶联法不同。This example provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which is basically the same as the method in Example 1, except that the urease coupling method in Step 1 is different.
尿素酶偶联法的具体步骤为：步骤a、试剂与血清混合，并于38℃温浴3min。其中，试剂由磷酸盐缓冲液70nmol/L、尿素酶6000U/L、谷氨酸脱氢酶10000U/L、α-酮戊二酸180nmol/L、NADH4.0nmol/L、防腐剂250μl/L组成。其中，磷酸盐缓冲液为TRIS-盐酸缓冲剂，防腐剂为ProcIin-300。血清与试剂的体积比为1:80。步骤b、在340nm波长处测量试剂和血清的混合液中NADH的吸光度下降速率，并与相同处理的尿素氮标准液比较，计算获得样品中尿素氮的含量。The specific steps of the urease coupling method are as follows: step a, mixing the reagent and serum, and incubating at 38° C. for 3 minutes. Among them, the reagent is composed of phosphate buffer 70nmol/L, urease 6000U/L, glutamate dehydrogenase 10000U/L, α-ketoglutarate 180nmol/L, NADH 4.0nmol/L, preservative 250μl/L . Wherein, the phosphate buffer is TRIS-hydrochloric acid buffer, and the preservative is Proclin-300. The volume ratio of serum to reagent is 1:80. Step b. Measure the decrease rate of absorbance of NADH in the mixture of reagent and serum at a wavelength of 340nm, and compare it with the same treated urea nitrogen standard solution to calculate the content of urea nitrogen in the sample.
其中，尿素氮标准液的浓度为8mmol/L。处理方式与实施例一相同。Among them, the concentration of urea nitrogen standard solution is 8mmol/L. The processing method is the same as that in Embodiment 1.
实施例三Embodiment three
本实施例提供一种血清中尿素氮含量近红外光谱测定方法，与实施例一的方法基本相同，区别仅在于：步骤一中的尿素酶偶联法不同。This example provides a method for measuring urea nitrogen content in serum by near-infrared spectroscopy, which is basically the same as the method in Example 1, except that the urease coupling method in Step 1 is different.
尿素酶偶联法的具体步骤为：步骤a、试剂与血清混合，并于37℃温浴4min。其中，试剂由尿素酶5000U/L、谷氨酸脱氢酶9000U/L、α-酮戊二酸160nmol/L、NADH3.0nmol/L。血清与试剂的体积比为1:75。步骤b、在340nm波长处测量试剂和血清的混合液中NADH的吸光度下降速率，并与相同处理的尿素氮标准液比较，计算获得样品中尿素氮的含量。The specific steps of the urease coupling method are as follows: step a, mixing the reagent and serum, and incubating at 37° C. for 4 minutes. Among them, the reagents are urease 5000U/L, glutamate dehydrogenase 9000U/L, α-ketoglutarate 160nmol/L, NADH 3.0nmol/L. The volume ratio of serum to reagent is 1:75. Step b. Measure the decrease rate of absorbance of NADH in the mixture of reagent and serum at a wavelength of 340nm, and compare it with the same treated urea nitrogen standard solution to calculate the content of urea nitrogen in the sample.
其中，尿素氮标准液的浓度为7mmol/L。处理方式与实施例一相同。Among them, the concentration of urea nitrogen standard solution is 7mmol/L. The processing method is the same as that in Embodiment 1.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107677634A (en) * | 2017-10-11 | 2018-02-09 | 合肥工业大学 | Method based on clenbuterol hydrochloride in Ftir Spectroscopy detection pig urine |
| CN108061708A (en) * | 2017-12-14 | 2018-05-22 | 青岛顺昕电子科技有限公司 | Urea content analysis system and its control system, control method |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2371982Y (en) * | 1999-06-08 | 2000-04-05 | 中国人民解放军北京军区总医院 | Ascitic fluid ultrafilter therapeutic equipment with double peristaltic pump |
| CN103276052A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Urea nitrogen detection reagent |
| CN105891150A (en) * | 2016-05-10 | 2016-08-24 | 广东星创众谱仪器有限公司 | Liquid detection device and detection method thereof for near-infrared spectrum analyzer |
| CN105950704A (en) * | 2016-04-26 | 2016-09-21 | 天津市宝坻区人民医院 | Method for simultaneous determination of double items of urea nitrogen and creatinine in serum |
| CN205656125U (en) * | 2016-05-25 | 2016-10-19 | 广东星创众谱仪器有限公司 | A liquid detection device for near infrared spectroscopy analysis appearance |
| CN106252196A (en) * | 2015-06-03 | 2016-12-21 | 东京毅力科创株式会社 | Substrate board treatment and substrate processing method using same |
| CN106323876A (en) * | 2011-01-21 | 2017-01-11 | 提拉诺斯公司 | Systems and methods for sample use maximization |
-
2017
- 2017-04-26 CN CN201710283945.2A patent/CN106908411A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2371982Y (en) * | 1999-06-08 | 2000-04-05 | 中国人民解放军北京军区总医院 | Ascitic fluid ultrafilter therapeutic equipment with double peristaltic pump |
| CN106323876A (en) * | 2011-01-21 | 2017-01-11 | 提拉诺斯公司 | Systems and methods for sample use maximization |
| CN103276052A (en) * | 2013-05-24 | 2013-09-04 | 宁波美康生物科技股份有限公司 | Urea nitrogen detection reagent |
| CN106252196A (en) * | 2015-06-03 | 2016-12-21 | 东京毅力科创株式会社 | Substrate board treatment and substrate processing method using same |
| CN105950704A (en) * | 2016-04-26 | 2016-09-21 | 天津市宝坻区人民医院 | Method for simultaneous determination of double items of urea nitrogen and creatinine in serum |
| CN105891150A (en) * | 2016-05-10 | 2016-08-24 | 广东星创众谱仪器有限公司 | Liquid detection device and detection method thereof for near-infrared spectrum analyzer |
| CN205656125U (en) * | 2016-05-25 | 2016-10-19 | 广东星创众谱仪器有限公司 | A liquid detection device for near infrared spectroscopy analysis appearance |
Non-Patent Citations (1)
| Title |
|---|
| 刘永峰 等: "应用近红外光谱法定量检测牛奶中尿素氮的研究", 《西北农林科技大学学报（自然科学版）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107677634A (en) * | 2017-10-11 | 2018-02-09 | 合肥工业大学 | Method based on clenbuterol hydrochloride in Ftir Spectroscopy detection pig urine |
| CN108061708A (en) * | 2017-12-14 | 2018-05-22 | 青岛顺昕电子科技有限公司 | Urea content analysis system and its control system, control method |
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