DETERMINATION OF UREA BY PHTHALDEHYDE METHOD
Release time:
2022-10-18
尿素是最早被测定的体液中的物质之一t其测定方 法主要有二乙酰乙肟法、脲酶法、酶偶联速率法等t这些方法或是存在煮沸，或是试剂价格较昂贵 目前，市场上
供应了邻苯二甲醛显色法(OPA)测定尿素试剂盒，该 法不需煮沸，简便、快速、准确、稳定易得。我们根据反应 原理，在 ISP-M 半自动生化分析仪上进行尿素测定， 效果满意。特做报道
1 原理
在酸性环境中，血清中的尿素在_妾替比林存在下与 邻苯二甲醛作用生成黄色物质，该物质在偏钒酸根的作 用下变为蓝色物质。该物质藩液颜色的深浅在一定范围
内与血清尿素浓度成正比，与同样处理的标准物一起比 色测定，即可得到血清标本中尿素的浓度。
2 材料与方法
2．I 试剂 OPA法测定尿索试剂盒}其中包括A液 B 液，尿素标准液，僳定长城临床试剂公司t批号 951102。 酶偶联速率法测定尿素试剂盒：上海长征医学科学有限
公司，批号 950828
2．2 仪器
IsP—M 半自动生化分析仅，型号V／talob 一 21。
2．3 方法 A液和 B液在临用前 10：1混台．25ul样 本中加入1．1ml混台液，37"(315分钟．在IsP—M半自 动 生化分析仪上 测定 终点法，比空白调霉，波长 600m'nt比色即可得出尿素的含量。同时以咎氨酰脱氢 醇偶联反应法，也在半自动生化分析仅上进行动力学分 析。
3 结果与讨论
作者单位，五河县压院检验科(233309) 试剂。但进口试剂价格较贵，使用国产试剂或自配试剂 必须达到与进口试剂相同的检验效果，井用标本质控液 进行标定，方可使用 丛玉隆在血液分析仪应用中的几个问题Ⅲ一文中 指出：细胞直方图既给临床提供诊断参考数据，也为操 作人员提供对仪器工作状态和实验结果是否可信的监 控，一般必须在仔细分析直方图后，确定是否需要显微 镜检查再发报告。我们应该不断的总结经验 ，发现问题， 不断提高专业人员的技术水平，使其更好的为临床医疗 工作服务
3．1 线性范围 在 0～71．4mraol／I范围内配制 Lo种 不同浓度的尿素溶液 ，每种浓度溶液重复测定 3捷 该 法在尿素维度o～28mraol／I之阃线性良好。
3．2 精密度实验 选用高、中、低值尿素(浓度为 Lo． 71、5．36、357mmol／I)，测定 2O次，其批内CV分别为 31％、23 、2．1’6，其批同 CV分别为 4．2％、3．6 、 3．2％ 。
3．3 回收事实验
在尿素浓度为 7．87和 21．50mmol／ l的血清标本中，加入尿素浓度为 6．Ommol／1．，然后进行 测定，其回收率分别为 90 和 98
3．4 干扰实验 配制 513mmol／1胆红索、7．6retool／ 1CHO、3．5mmol／1甘油三酯、4gA血红蛋白做干扰实 验．发现无明显干扰 (P>005)t维生索 C浓度低于
75mg／1时无干扰(P>0．05)实验室使用的重氨试剂对 苯磺酸可干扰反应．测定时应防止此类物质在实验室中 污染 。
3．5 准确性实验 用 OPA显色法和各氧酰脱氢酶偶联速率法同时测定血清标本 50倒(本院门诊健康体检 者)，分别得 土S为 5．18士1．70、5．28±1．73。统计学
处理，两法无显著性差异(P>0．05)，相关系数 r为 o． 9804．回归方程 Y一1．0ix+0．0456。
总之，邻苯二甲醛法测定尿索既有酶法的方便．可 自动化分析，又价廉，试剂稳定可室温保存，且变异小干 扰固索少，线性范围良好，并与酶法有高度相关。实践证
明该法不失为经济、实用值得推广的尿素测定技术。
Urea is one of the first substances to be measured in body fluids.Master of LawThere should be diacetyl glyoxime method, urease method, enzyme coupling rate method, etc.The method is either boiling, or the reagent is more expensive at present, the market.
The supply of phthalaldehyde chromogenic method (OPA) determination of urea kit,The method does not need to boil, is simple, rapid, accurate, stable and easy to obtain. We react accordingprinciple, in the ISP-Urea determination on M semi-automatic biochemical analyzer,The effect is satisfied. Special report
1 Principle
In an acidic environment, urea in serum in the presence of_concubineThe action of o-phthalaldehyde generates a yellow substance, which is used in the production of metavanadate.Use it to turn into a blue substance. The depth of the color of the substance's liquid is within a certain range.
It is proportional to the serum urea concentration and is compared with the standard of the same treatment.The concentration of urea in the serum sample can be obtained by color determination.
2 Materials and methods
2.I reagent OPA assay urine cord kit} which includes liquid a BLiquid, urea standard solution, batch number 951102 of t of bo ding great wall clinical reagent co., ltd.Determination of urea by enzyme coupling rate method kit: Shanghai Changzheng Medical Science Co., Ltd.
Company, lot number 950828
2.2 instrument
IsP-M semi-automatic biochemical analysis only, model V/talobOne21。
2.3 Method Liquid A and Liquid B are mixed 10:1 before use. 25ul sampleAdd 1.1ml of mixed solution to this, 37"(3Fifteen minutes. In IsP-M semi-selfDetermination of end point method on dynamic biochemical analyzer, than blank mold, wavelength600m'The content of urea can be obtained by nt colorimetry. At the same time to blame amino acyl dehydrogenationAlcohol coupling reaction method, also in semi-automatic biochemical analysis only on dynamic creditsAnalysis.
3 Results and discussion
Author Unit, Inspection Department of Wuhe County Press Institute (233309)Reagents. But the price of imported reagents is more expensive, the use of domestic reagents or self-made reagents.Must achieve the same inspection effect as imported reagents, well-used sample quality control fluid.Calibration can be used beforeSeveral Problems of Cong Yulong in the Application of Hematology Analyzer ⅢIt is pointed out that the cell histogram not only provides diagnostic reference data for clinical, but also for the operation.The staff shall provide monitoring of the working status of the instrument and the reliability of the experimental results.control, it is generally necessary to determine whether microscopy is required after careful analysis of the histogram.The mirror examination report was sent again. We should continue to sum up experience and identify problems,Continuously improve the technical level of professionals to make it better for clinical treatment.Work Service
3.1 linear range in the range of 0~71.4mraol/I for the preparation of Lo speciesDifferent concentrations of urea solution, each concentration solution repeated determination of 3method in the urea dimension o ~ 28mraol/I has good linearity.
High, medium and low value urea (concentration of Lo．71, 5.36, 357mmol/I), measured 2O times, the batch CV is respectively31%, 23, 2.1 '6, with batch-to-batch CV of 4.2, 3.6,3．2％ 。
3.3 Recovery Experiment
at urea concentrations of 7.87 and 21.50mmol/In the serum sample of l, add urea at a concentration of 6.0 mmol/1., and then proceeddetermination, the recoveries were 90 and 98
Preparation of 513mmol/1 bile red cord and 7.6r in 3.4 interference experimentetool／1CHO, 3.5mmol/1 triglyceride, 4gA hemoglobin do interferenceCheck. It was found that there was no obvious interference (P>005)t-dimensional cord C concentration was lower
75mg/1 without interference (P>0.05) laboratory use of heavy ammonia reagent pairBenzenesulfonic acid can interfere with the reaction. Such substances should be prevented from being measured in the laboratory.Pollution.
OPA chromogenic method and each oxyacyl dehydrogenase couple for 3.5 accuracy experimentSimultaneous determination of serum specimens by combined rate method 50 inverted (hospital outpatient health examination), respectively, soil s is 5.18 1.70, 5.28±1.73. Statistics
treatment, there is no significant difference between the two methods (P. 0.05), and the correlation coefficient r is o.．9804. Regression equation Y a 1.0ix 0.0456.
In a word, the determination of urine by the o-phthalaldehyde method is convenient for the enzymatic method. CanAutomated analysis, inexpensive, stable reagents can be stored at room temperature, and the variation is small and dry.The linear range is good and highly correlated with enzymatic methods. Practice Certificate
This method is an economical and practical urea determination technique worthy of promotion.
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