本文關(guān)鍵詞：重氮偶聯(lián)分光光度法測(cè)定人體血液中亞硝酸鹽含量的比較研究及亞硝酸鹽在大鼠主要檢材中的分布　出處：《昆明醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 亞硝酸鹽 紫外分光光度法 比較研究 對(duì)氨基苯磺酸 4-氨基-5-羥基萘-2 7-二磺酸單鈉鹽

【摘要】：目的 選擇六組具有代表性的重氮偶聯(lián)分光光度法測(cè)定人體血液中亞硝酸鹽含量的最佳偶合劑,探索各組重氮偶合劑的最適反應(yīng)條件,建立六組人體血液中亞硝酸鹽的分光光度計(jì)定量分析方法；用所建立的方法,分別測(cè)出六組偶聯(lián)-分光光度法測(cè)定亞硝酸鹽含量的線性范圍、檢出限、人體血液中亞硝酸鹽的含量和各組方法的精密度及回收率,比較不同試劑重氮偶聯(lián)測(cè)定血液中亞硝酸鹽含量的優(yōu)越性,最終確定最適宜分析生物檢材中亞硝酸鹽含量的重氮偶聯(lián)分光光度法。 建立急性亞硝酸鹽中毒大鼠模型,選用最適宜分析生物檢材中亞硝酸鹽含量的重氮偶聯(lián)分光光度法,即對(duì)氨基苯磺酸溶液與4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液分別為重氮試劑和偶聯(lián)試劑測(cè)定亞硝酸鹽中毒大鼠模型心血、肝臟、胃及內(nèi)容物和腎臟中亞硝酸鹽的含量,對(duì)實(shí)驗(yàn)數(shù)據(jù)進(jìn)行分析,闡明急性亞硝酸鹽中毒大鼠主要臟器中亞硝酸鹽的分布狀況。 方法 1.通過查閱文獻(xiàn),選定六組具有代表性的偶聯(lián)-分光光度法測(cè)定亞硝酸鹽含量的試劑方法：(1)對(duì)氨基苯磺酰胺和鹽酸萘乙二胺分別為重氮試劑和偶氮試劑；(2)對(duì)氨基苯磺酸和8-羥基喹啉分別為重氮試劑和偶氮試劑；(3)對(duì)氨基苯磺酸和4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽分別為重氮試劑和偶氮試劑；(4)對(duì)氨基苯磺酸和1-苯基-3-甲基-5-吡唑啉酮分別為重氮試劑和偶氮試劑；(5)4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽(H酸)既作為重氮試劑又作為偶氮試劑(偶聯(lián)反應(yīng)在pH7.5條件下進(jìn)行)；(6)4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽既作為重氮試劑又作為偶氮試劑(偶聯(lián)反應(yīng)在pH4.5條件下進(jìn)行)。 2.隨機(jī)抽取10個(gè)健康成年人靜脈血各1mL混合(內(nèi)含抗凝劑),抽取1mL混合血置于250mL容量瓶中,加入50mL超純水和10mL氫氧化鈉溶液(20.0g/L),再加入10mL硫酸鋅溶液(0.42mol/L),混合均勻。置60℃水浴中加熱10min,取出冷卻至室溫,定容至250mL,混合均勻。放置30min,使用定量濾紙過濾并棄去初濾液50mL,加入干燥的亞硝酸鈉160μg,使之完全溶解。 3.每組試劑方法分別固定其它反應(yīng)條件,改變一種反應(yīng)條件,測(cè)其最適宜的反應(yīng)參數(shù),確定總反應(yīng)最適宜條件；建立六組試劑方法,測(cè)定每組方法的相關(guān)參數(shù),分別用六組方法測(cè)定血液中亞硝酸鹽含量及回收率,并進(jìn)行六組方法結(jié)果的比較研究。 4.將14只健康雄性Sprague-Dawley大鼠隨機(jī)分為空白對(duì)照組(n=4)、生理鹽水灌胃對(duì)照組(n=4)和亞硝酸鈉灌胃實(shí)驗(yàn)組(n=6)。分別對(duì)每組動(dòng)物進(jìn)行編號(hào)。生理鹽水對(duì)照組和實(shí)驗(yàn)組大鼠均灌胃給藥�？瞻讓�(duì)照組采用斷頭法處死,生理鹽水對(duì)照組于給藥后60min采用斷頭法處死,實(shí)驗(yàn)組在給藥后任其死亡�？焖偬崛∷写笫笮难⒏闻K、胃及內(nèi)容物和腎臟,以超純水浸泡萃取其中的亞硝酸鹽。 5.以對(duì)氨基苯磺酸溶液與4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液分別為重氮試劑和偶聯(lián)試劑,采用標(biāo)準(zhǔn)工作曲線法測(cè)定各組大鼠心血、肝臟、胃及內(nèi)容物和腎臟中亞硝酸鹽的含量,采用SPSS軟件對(duì)定量分析結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果 1.對(duì)氨基苯磺酰胺溶液與鹽酸萘乙二胺溶液分別為重氮試劑和偶聯(lián)試劑反應(yīng)條件：室溫下,在10mL比色管中加入4.0g/L pH1鹽酸對(duì)氨基苯磺酰胺溶液1mL及梯度濃度亞硝酸鈉溶液1mL,靜置3min進(jìn)行重氮反應(yīng),后加入1.0g/L鹽酸萘乙二胺溶液1mL,加水至刻度,混勻,靜置1min進(jìn)行偶合反應(yīng),以未加亞硝酸鈉的混合溶液為參比進(jìn)行吸光度測(cè)定。此反應(yīng)生成紫紅色的偶聯(lián)產(chǎn)物,在最大吸收波長(zhǎng)540nm處,亞硝酸鹽的濃度在0.1-1.51μg/mL范圍內(nèi)符合Beer定律,相關(guān)系數(shù)丫為0.99975,檢出限為3.86ng/mL; 2.對(duì)氨基苯磺酸溶液與8-羥基喹啉溶液分別為重氮試劑和偶聯(lián)試劑反應(yīng)條件：室溫下,在10mL比色管中加入4.0g/L pH1鹽酸對(duì)氨基苯磺酸溶液1mL及系列梯度濃度亞硝酸鈉溶液1mL,靜置1min進(jìn)行重氮反應(yīng),后加入0.6g/L8-羥基喹啉溶液1mL及pH12.5氨水1mL,加水至刻度,混勻,靜置1mmin進(jìn)行偶合反應(yīng),以未加亞硝酸鈉的混合溶液為參比進(jìn)行吸光度測(cè)定。此反應(yīng)生成桔黃色偶聯(lián)產(chǎn)物,在最大吸收波長(zhǎng)500nm處,亞硝酸鹽的濃度在0.2-4.6μg/mL范圍內(nèi)符合Beer定律,相關(guān)系數(shù)γ為0.99995,檢出限為26.13ng/mL; 3.對(duì)氨基苯磺酸溶液與4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液分別為重氮試劑和偶聯(lián)試劑反應(yīng)條件：室溫下,在10mL比色管中加入1.0g/L pH1鹽酸對(duì)氨基苯磺酸溶液1mL及系列梯度濃度亞硝酸鈉溶液1mL,靜置3mmin進(jìn)行重氮反應(yīng),后加入1.0g/L4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液1mL及pH4.5醋酸-醋酸鈉緩沖液1mL,加水至刻度,混勻,靜置30min進(jìn)行偶合反應(yīng),以未加亞硝酸鈉的混合溶液為參比進(jìn)行吸光度測(cè)定。此反應(yīng)生成粉紅色偶聯(lián)產(chǎn)物,在最大吸收波長(zhǎng)520nm處,亞硝酸鹽的濃度在0.2-5.25μg/mL范圍內(nèi)符合Beer定律,相關(guān)系數(shù)γ為0.99970,檢出限為61.58ng/mL； 4.對(duì)氨基苯磺酸溶液與1-苯基-3-甲基-5-吡唑啉酮溶液分別為重氮試劑和偶聯(lián)試劑反應(yīng)條件：室溫下,在10mL比色管中加入8g/L pH0.45鹽酸對(duì)氨基苯磺酸溶液1mL及系列梯度濃度亞硝酸鈉溶液1mL,靜置1min進(jìn)行重氮反應(yīng),后加入pH7.5磷酸鹽緩沖溶液1mL及1.0g/L1-苯基-3-甲基-5-吡唑啉酮溶液1mL,加水至刻度,混勻,靜置20min進(jìn)行偶合反應(yīng),以未加亞硝酸鈉的混合溶液為參比進(jìn)行吸光度測(cè)定。此反應(yīng)生成金黃色偶聯(lián)產(chǎn)物,在最大吸收波長(zhǎng)390nm處,亞硝酸鹽的濃度在0.2-6.0μg/mL范圍內(nèi)符合Beer定律,相關(guān)系數(shù)γ為0.99998,檢出限為15.13ng/mL; 5.4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽在pH7.5磷酸鹽緩沖液條件下與亞硝酸鹽反應(yīng)反應(yīng)條件：室溫下,在10mL比色管中加入8g/L pH1鹽酸4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液1mL及系列梯度濃度亞硝酸鈉溶液1mL,靜置3min進(jìn)行重氮反應(yīng),后加入pH7.5磷酸鹽緩沖溶液(磷酸二氫鈉-磷酸氫二鈉緩沖溶液)1mL,加水至刻度,混勻,靜置6min進(jìn)行偶合反應(yīng),以未加亞硝酸鈉的混合溶液為參比進(jìn)行吸光度測(cè)定。此反應(yīng)生成紫紅色偶聯(lián)產(chǎn)物,在最大吸收波長(zhǎng)550nm處,亞硝酸鹽的濃度在0.8-7.25μg/mL范圍內(nèi)符合Beer定律,相關(guān)系數(shù)γ為0.99081,檢出限為632.89ng/mL; 6.4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽在pH4.5醋酸鹽緩沖液(醋酸-醋酸鈉緩沖溶液)條件下與亞硝酸鹽反應(yīng)反應(yīng)條件：55℃水浴條件下,在10mL比色管中加入6.0g/LpH1鹽酸4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液1mL及系列梯度濃度亞硝酸鈉溶液1mL靜置3min進(jìn)行重氮反應(yīng),后加入pH4.5醋酸-醋酸鈉緩沖液1mL,加水至刻度,混勻,靜置6min進(jìn)行偶合反應(yīng),以未加亞硝酸鈉的混合溶液為參比進(jìn)行吸光度測(cè)定。此反應(yīng)生成紫紅色偶聯(lián)產(chǎn)物,在最大吸收波長(zhǎng)550nm處,亞硝酸鹽的濃度在0.4-5.25μg/mL范圍內(nèi)符合Beer定律,相關(guān)系數(shù)γ為0.99930,檢出限為193.99ng/mL。 7.除4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽(偶氮反應(yīng)在pH7.5條件下完成)方法,其他五種方法靈敏度高且線性好。對(duì)氨基苯磺酸和4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽分別為重氮組分和偶氮組分、對(duì)氨基苯磺酸和1-苯基-3-甲基-5-吡唑啉酮分別為重氮組分和偶氮組分、4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽(偶氮反應(yīng)在pH7.5條件下完成)和H酸(偶氮反應(yīng)在pH4.5條件下完成)四種方法線性范圍上限高,可測(cè)較大濃度亞硝酸鹽含量；對(duì)氨基苯磺酰胺和鹽酸萘乙二胺分別為重氮組分和偶氮組分方法線性范圍下限低,可測(cè)痕量亞硝酸鹽的含量。對(duì)氨基苯磺酸和8-羥基喹啉分別為重氮組分和偶氮組分、對(duì)氨基苯磺酸和4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽分別為重氮組分和偶氮組分、對(duì)氨基苯磺酸和1-苯基-3-甲基-5-吡唑啉酮分別為重氮組分和偶氮組分和4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽(偶氮反應(yīng)在pH4.5條件下完成)方法穩(wěn)定性較好。對(duì)氨基苯磺酸和1-苯基-3-甲基-5-吡唑啉酮分別為重氮組分和偶氮組分方法測(cè)得的血液亞硝酸鹽回收率最高,對(duì)氨基苯磺酸和4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽分別為重氮組分和偶氮組分方法的回收率次之,但測(cè)定的人體血液中亞硝酸鹽含量最接近加入量,準(zhǔn)確度最好。 8.對(duì)照組大鼠未見任何不良反應(yīng),實(shí)驗(yàn)組大鼠在瀕死期均出現(xiàn)皮膚黏膜發(fā)紺和呼吸困難等癥狀,且均在90min內(nèi)死亡。經(jīng)對(duì)氨基苯磺酸和4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽分別為重氮組分和偶氮組分方法檢測(cè),空白對(duì)照組和生理鹽水組大鼠檢材中均未測(cè)出亞硝酸鹽。亞硝酸鈉給藥實(shí)驗(yàn)組測(cè)得胃及內(nèi)容物中亞硝酸鈉含量最多,血液中含量次之,肝臟中亞硝酸鈉含量最少。 結(jié)論 對(duì)氨基苯磺酸溶液與4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液分別為重氮試劑和偶聯(lián)試劑方法,穩(wěn)定性較好,線性范圍廣,測(cè)量人體血液亞硝酸鹽含量實(shí)驗(yàn)中,其準(zhǔn)確度及回收率較高,最適合測(cè)定生物檢材中亞硝酸鹽的含量。 檢測(cè)急性亞硝酸鈉中毒案例最好的生物檢材為胃及內(nèi)容物和心血,在今后司法檢測(cè)鑒定亞硝酸鹽中毒的案例中可采用對(duì)氨基苯磺酸溶液與4-氨基-5-羥基萘-2,7-二磺酸單鈉鹽溶液分別為重氮試劑和偶聯(lián)試劑方法。
[Abstract]:objective
Six was the best coupling agent Determination of nitrite content in human blood diazo coupling representative spectrophotometry, the optimum reaction conditions were explored diazo coupling agent, spectrophotometry to establish six groups of nitrite in human blood count quantitative analysis method; with the establishment of the six groups were analyzed. Coupled spectrophotometric determination of nitrite content in the linear range, detection limit, precision and recovery of each method and content of nitrite in human blood rate, comparative superiority of determination of nitrite content in the blood of the diazo coupling reagent, and ultimately determine the most suitable diazo coupling analysis of nitrite content in biological samples by spectrophotometry spectrophotometric method.
Establishment of a rat model of acute nitrite poisoning, choose the most suitable diazo coupling analysis of nitrite content in biological samples by spectrophotometry, namely p-aminobenzenesulfonic acid solution and 4- amino -5- -2,7- two hydroxy naphthalene sulfonic acid monosodium salt solution respectively as diazo reagent and coupling reagent determination of nitrite poisoning rat model of blood, liver, content the nitrite contents of stomach and kidney and the analysis of experimental data, to clarify the distribution of nitrite in major organs of rats with acute nitrite poisoning.
Method
1. through the literature, selected six groups of reagent method determination of nitrite content of coupling - representative spectrophotometric method: (1) Sulfanilamide and hydrochloric Naphthylethylenediamine respectively diazo and azo reagents; (2) sulfanilate and 8- hydroxyquinoline were diazo and azo reagents (3); p-aminobenzenesulfonic acid and 4- amino -5- hydroxy naphthalene sulfonic acid monosodium salt -2,7- two respectively, diazo and azo reagents; (4) sulfanilic acid and 1- phenyl -3- methyl -5- pyrazolone respectively diazo and azo reagents; (5) 4- -5- -2,7- amino hydroxy naphthalene two sulfonic acid monosodium salt (H) as diazo and azo reagents (as coupling reaction under the condition of pH7.5); (6) 4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt as diazo and azo reagents (as coupling reaction under the condition of pH4.5).
2. randomly selected 10 healthy adults 1mL mixed venous blood (containing anticoagulants), mixed blood from 1mL to a 250mL volumetric flask, add 50mL ultrapure water and 10mL sodium hydroxide solution (20.0g/L), adding 10mL solution of zinc sulfate (0.42mol/L), mixed evenly. The heating 10min 60 C water bath, remove the cooling to room temperature and set the volume at 250mL, mixed evenly. Placed 30min, using quantitative filter and discard primary filtrate 50mL, adding dry sodium nitrite 160 G, which is completely dissolved.
3. each reagent method respectively fixed other reaction conditions, a change in the reaction conditions, the optimum reaction parameters, determine the total reaction the most suitable conditions; establish six group reagent method, related parameters determination method in each group, respectively with six sets of methods for determination of nitrite content and recovery rate of blood, comparative study and six the results of the group method.
4. a total of 14 healthy male Sprague-Dawley rats were randomly divided into control group (n=4), saline control group (n=4) and sodium nitrite perfused experimental group (n=6) respectively. The number of each animal. The saline control group and experimental group rats were intragastric administration of blank. The control group were killed by decapitation, the saline control group after administration by 60min were killed by decapitation, the experimental group in the administration after its death. The rapid extraction of all rats blood, liver, stomach and kidney and the contents of nitrite, ultra pure water for extraction of them.
5. with p-aminobenzenesulfonic acid solution and 4- amino -5- -2,7- two hydroxy naphthalene sulfonic acid monosodium salt solution respectively as diazo reagent and coupling reagent, determination of blood, the rats liver by standard working curve method, and the contents of nitrite content in the stomach and kidney, the quantitative analysis results were statistically analyzed by SPSS software.
Result
1. sulfanilamide solution and hydrochloric acid naphthalene ethylenediamine respectively diazo reagent and coupling reagent reaction conditions: room temperature, 10mL colorimetric tube and add 4.0g/L pH1 hydrochloride on 1mL and concentration gradient BENZENESULFONAMIDES solution of sodium nitrite solution 1mL, static 3min diazo reaction, after adding 1.0g/L hydrochloric acid naphthalene ethylenediamine 1mL, add water to the scale, mixing, static 1min coupling reaction, the reference absorbance was determined by mixed solution without nitrite. This reaction produces coupling product purple, the maximum absorption wavelength of 540nm, the concentration of nitrite in the range of 0.1-1.51 in the range of g/mL accords with Beer law. The correlation coefficient y is 0.99975, the detection limit is 3.86ng/mL;
2. sulfanilic acid solution and 8- hydroxyquinoline solution respectively diazo reagent and coupling reagent reaction conditions at room temperature in the 10mL colorimetric tube into 4.0g/L pH1 hydrochloric acid p-aminobenzenesulfonic acid solution 1mL and series of concentrations of sodium nitrite solution 1mL, static 1min diazo reaction, after adding 0.6g/L8- hydroxyquinoline 1mL and pH12.5 1mL ammonia solution, add water to the scale, mixing, static 1mmin coupling reaction, the reference absorbance was determined by mixed solution without nitrite. This reaction produces orange coupling product, the maximum absorption wavelength of 500nm, nitrite concentration in the range of g/mL 0.2-4.6 with Beer related law the coefficient is 0.99995, the detection limit is 26.13ng/mL;
3. sulfanilic acid solution with 4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt solution respectively as diazo reagent and coupling reagent reaction conditions at room temperature in the 10mL colorimetric tube into 1.0g/L pH1 hydrochloric acid p-aminobenzenesulfonic acid solution 1mL and series of concentrations of sodium nitrite solution 1mL, static 3mmin diazo after the reaction, adding 1.0g/L4- amino -5- hydroxy naphthalene sulfonic acid sodium salt solution two -2,7- single 1mL and pH4.5 acetic acid sodium acetate buffer 1mL, add water to the scale, mixing, static 30min coupling reaction, the reference of absorbance was determined by mixed solution without nitrite. This reaction produces pink coupling products. The maximum absorption wavelength of 520nm, nitrite concentration in the range of g/mL 0.2-5.25 with Beer law, related coefficient is 0.99970, the detection limit is 61.58ng/mL;
4. sulfanilic acid solution with 1- phenyl -3- methyl -5- pyrazolone solution respectively for diazo reagent and coupling reagent reaction conditions at room temperature in the 10mL colorimetric tube into 8g/L pH0.45 hydrochloric acid p-aminobenzenesulfonic acid solution 1mL and series of concentrations of sodium nitrite solution 1mL, static 1min diazo reaction after joining pH7.5, 1mL phosphate buffer and 1.0g/L1- phenyl -3- methyl -5- pyrazolone solution 1mL, add water to the scale, mixing, static 20min coupling reaction, the reference absorbance was determined by mixed solution without nitrite. This reaction produces gold coupling products in the maximum absorption wavelength of 390nm 0.2-6.0, the nitrite concentration in the range of g/mL with the Beer law, related coefficient is 0.99998, the detection limit is 15.13ng/mL;
5.4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt in pH7.5 phosphate buffer conditions and nitrite reaction conditions: room temperature, 10mL colorimetric tube and add 8g/L pH1 4- -5- -2,7- hydrochloride amino hydroxy naphthalene sulfonic acid sodium salt solution and two single 1mL series of concentrations of sodium nitrite solution 1mL, static 3min weight nitrogen reaction after adding pH7.5 phosphate buffer solution (sodium dihydrogen phosphate - two sodium hydrogen phosphate buffer solution) 1mL, add water to the scale, mixing, static 6min coupling reaction, the reference absorbance was determined by mixed solution without nitrite. This reaction produces purple coupling products in the maximum absorption wavelength of 550nm 0.8-7.25, the nitrite concentration in the range of g/mL with the Beer law, related coefficient is 0.99081, the detection limit is 632.89ng/mL;
6.4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt in pH4.5 acetate buffer (sodium acetate buffer solution) under the condition of nitrite and reaction conditions: 55 DEG C water bath under the condition of 10mL in the cuvette adding 6.0g/LpH1 HCl 4- amino -5- hydroxy naphthalene sulfonic acid sodium salt solution two -2,7- single 1mL and a series of sub concentration gradient sodium nitrate solution 1mL static 3min diazo reaction, after adding pH4.5 acetic acid sodium acetate buffer 1mL, add water to the scale, mixing, static 6min coupling reaction, the reference absorbance was determined by mixed solution without nitrite. This reaction produces purple coupling products at the maximum absorption wavelength at 550nm, the nitrite concentration in the range of g/mL 0.4-5.25 with Beer law, related coefficient is 0.99930, the detection limit is 193.99ng/mL.
In addition to the 7. 4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt (azo reaction under the condition of pH7.5), the other five methods with high sensitivity and good linearity. Sulfanilic acid and 4- amino -5- hydroxy naphthalene sulfonic acid monosodium salt -2,7- two respectively as diazo component and azo group, sulfanilic acid and 1- phenyl -3- methyl -5- pyrazolone respectively as diazo component and azo group, 4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt (azo reaction under the condition of pH7.5) and H (azo reaction was completed under the condition of pH4.5) four methods of linear range limit, measurable greater concentration of nitrite content; Sulfanilamide and hydrochloric Naphthylethylenediamine respectively as diazo component and azo group method is linear in the range of lower limit, we can measure the contents of trace nitrite. P-aminobenzenesulfonic acid and 8- hydroxyquinoline respectively as diazo component and azo group, 4-aminobenzene 4- -5- amino acid and hydroxy naphthalene sulfonic acid monosodium salt -2,7- two respectively as diazo component and azo group, sulfanilic acid and 1- phenyl -3- methyl -5- pyrazolone respectively as diazo component and azo group and 4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt (azo reaction in pH4.5 under the condition of stable method.) sulfanilic acid and 1- phenyl -3- methyl -5- pyrazolone respectively as diazo component and azo group method measured the blood nitrite recovery rate was highest, sulfanilic acid and 4- amino -5- hydroxy naphthalene sulfonic acid monosodium salt -2,7- two respectively as diazo component and azo composition for the recovery time, but the determination of nitrite content in human blood is most close to the amount of the best accuracy.
8. control rats without any adverse reactions, the experimental group was observed in rat skin mucous membrane cyanosis and dyspnea and other symptoms in the dying period, and all died within 90min. By p-aminobenzenesulfonic acid and 4- amino -5- hydroxy naphthalene sulfonic acid monosodium salt -2,7- two respectively as diazo component and azo component method, the control group and the saline group rats samples were not detected in nitrite. Sodium nitrite administered experimental group measured gastric contents and most nitrite content in the blood content of sodium nitrite content in the liver at least.
conclusion
P-aminobenzenesulfonic acid solution with 4- -5- -2,7- two amino hydroxy naphthalene sulfonic acid monosodium salt solution respectively as diazo reagent and coupling reagent method, good stability, wide linear range, measurement of human blood nitrite content in the experiment, the accuracy and recovery rate is high, the most suitable for the determination of nitrite content in biological samples.
Acute sodium nitrite poisoning cases detected the best biological samples for the stomach and the contents and efforts, in case of nitrite poisoning in judicial identification using p-aminobenzenesulfonic acid solution and 4- amino -5- -2,7- two hydroxy naphthalene sulfonic acid monosodium salt solution respectively as diazo reagent and coupling reagent method.

【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：D919.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 劉希東;間苯二酚偶聯(lián)反應(yīng)測(cè)定痕量亞硝酸根的光度法研究[J];渝西學(xué)院學(xué)報(bào)(自然科學(xué)版);2003年02期

，

本文編號(hào)：1375173