尿中马尿酸、甲基马尿酸的高效液相色谱测定法

目的建立高效液相色谱法测定尿中马尿酸和甲基马尿酸（2、3、4-甲基马尿酸）。方法选择甲醇＋0．4％醋酸水溶液为流动相，检测波长为254nm，流速0．7ml／min；样品处理后经高效液相反相分离，采用色谱保留时间定性，峰面积定量。结果在选定的色谱条件下，标准曲线的线性关系良好，其相关系数大于0．999；精密度试验，相对标准偏差为0．25％-2．36％（n=6）；样品加标回收率为93．37％-100．6％；方法最低检测浓度马尿酸为0．038mg／L，2-甲基马尿酸为0．12mg／L，3．甲基马尿酸为0．011mg／L，4．甲基马尿酸为0．026mg／L。结论该方法操作简单、方法精密度、准确度均能满足实验要求，结果可靠。     

尿中甲基马尿酸和马尿酸共存时薄层分离与比色测定法

本文报告了用薄层层析分离和对二甲基氨基苯甲醛比色测定尿中甲基马尿酸和马尿酸的方法。方法检测限0.004g/L(1.0ml尿)。尿样全程测定精密度:甲基马尿酸4～9.5%,马尿酸4.3～9.6%。样品加标回收率:甲基马尿酸83～103%,马尿酸86～92%     

高效液相色谱串联质谱法测定聚丙烯酰胺残留单体

目的 建立聚丙烯酰胺水处理剂及饮用水中丙烯酰胺的高效液相色谱-串联质谱测定方法。方法 聚丙烯酰胺处理剂直接用甲醇超声提取,水样通过活性炭固相萃取柱富集,甲醇洗脱,采用T3色谱柱分离,以甲醇-0.1%甲酸水溶液作为流动相,以多反应监测(multiple reaction monitoring,MRM),ESI⁺电离方式,外标法定量。结果 丙烯酰胺在1.0～100.0 μg/L和0.50～10.0 mg/L质量浓度范围内线性良好,相关系数分别为0.999 3和0.999 8,方法回收率为95.0%～102%,RSD≤4.7%,方法检出限分别为2 μg/kg和0.000 2 μg/L。结论 该方法快速、准确、灵敏,适合聚丙烯酰胺水处理剂和饮用水中丙烯酰胺的测定。     

高效液相色谱法测定尿中马尿酸的规范化研究

目的:进一步完善和规范化甲苯接触者尿中马尿酸的高效液相色谱测定法。方法:采用ODS C18柱,流动相为甲醇-0.01 mol/L磷酸(40+60),流量为1.0 ml/min,检测波长228 nm。结果:马尿酸在10μg/ml～500μg/ml范围内线性良好,相关系数r为0.9999,平均加标回收率为99%,平均相对标准偏差RSD为1.1%,最低检测浓度0.41 mg/L。结论:该方法快速、准确、灵敏度高,有良好的重现性和稳定性,可用于职业接触甲苯人群尿中马尿酸的测定。     

磁固相萃取-高效液相色谱检测牛肉样品中的游离雌激素

目的 建立磁固相萃取(magnetic solid phase extraction,MSPE)与高效液相色谱-二极管阵列检测器(high performance liquid chromatography-diode array detector,HPLC-DAD)检测牛肉中游离雌激素的新方法。 方法 制备基于金属有机骨架化合物的磁性纳米粒子,作为MSPE吸附材料与HPLC-DAD建立检测17β-雌二醇、雌酮、炔雌醇、己烯雌酚、双烯雌酚和双酚A的新方法,并将该方法应用于实际牛肉样品中这6种雌激素的分析检测。 结果 方法中6种雌激素的检出限为0.1~0.8 μg/L,线性范围为2~5 000 μg/L 和5~5 000 μg/L,方法标准偏差为4.9%~10.5%(n=8),实际样品加标回收率为79.1%~118%。 结论 新方法具有检出限低,线性范围广、抗干扰能力强等优点,对于检测生肉类样品中雌激素具有良好的应用。     

尿中马尿酸,甲基马尿酸的高效液相色谱测定法

本文在酸性下用醋酸乙酯提取,反相C_(18)柱,酸性甲醇/水系统,液相色谱法测定尿中马尿酸(1)和甲基马尿酸(2)。测定范围0.1～1.0mg/L(取1ml尿分析),检测限0.015mg/L(1),0.03m g/L(2)。变异系数(n=6)在5％以下。接触者尿加标平均回收率为99.05％(1)和104.3％(2)。尿样在4℃可保存1周;提取后蒸去溶剂的残渣在室温可保存半年。     

职业接触二甲苯尿中甲基马尿酸生物限值的研究

选择不同暴露水平的二甲苯接触个体,检测个体二甲苯暴露浓度,同时采集当日工人班末尿样,对尿中二甲苯的代谢产物甲基马尿酸的含量进行测定,将二甲苯的外暴露空气浓度与体内甲基马尿酸代谢量进行回归分析,结合国外限值资料,提出适合我国职业卫生现状的甲基马尿酸生物限值的建议。接触工人在空气中二甲苯浓度为0～144.21 mg/m3时,班末尿中甲基马尿酸(MHA)与空气中二甲苯浓度之间存在良好相关性,肌酐校正回归方程为y(g/g肌酐)=0.0052x(mg/m3)+0.0112,r=0.731;比重校正回归方程为y(g/L)=0.0089x(mg/m3)-0.0259,r=0.793。将我国二甲苯职业接触限值50 mg/m3分别代入回归方程,推出工作班末尿中甲基马尿酸含量为0.271 g/g肌酐和0.419 g/L。依据我国二甲苯实际接触情况,参考国外相关标准和文献,建议我国职业接触二甲苯的代谢产物尿中甲基马尿酸的生物接触限值为班末尿0.3 g/g肌酐或0.4 g/L。     

早产儿视网膜病患儿出生尿代谢产物差异分析的初步研究

目的 探讨早产儿视网膜病(ROP)患儿出生尿代谢产物特征性变化,以期为ROP的诊断提供新的生物标记物。方法 收集2013年1月—2016年12月于中山大学附属第六医院住院的21例ROP实验组患儿及同期21例非ROP对照组患儿尿液标本,利用气相色谱-质谱技术进行代谢产物测定,运用正交偏最小二乘判别分析法(OPLS-DA)进行代谢组学分析。结果 联合代谢聚类图与权重图分析发现10个权重较大的物质(油酸、犬尿酸、2-甲基3-羟基丁酸、3-氨基异丁酸、3-羟基3-甲基戊二酸、马尿酸、蛋氨酸、甘油酸、次黄嘌呤和4-羟基苯乳酸)中ROP组患儿尿甲硫氨酸、3-氨基异丁酸、马尿酸水平明显降低,3-羟基3-甲基戊二酸明显升高,与对照组比较差异均具有统计学意义(P均<0.05)。结论 尿甲硫氨酸、3-羟基-3-甲基戊二酸、3-氨基异丁酸、马尿酸等代谢产物在ROP早期诊断中具有潜在的应用价值。     

液相色谱-蒸发光散射检测法测定保健食品中8种糖的含量

目的 采用蒸发光散射检测器,建立高效液相色谱法检测保健食品中8种糖的含量。 方法 超声辅助加热溶剂萃取保健食品样品中的8种糖(木糖、果糖、山梨糖、葡萄糖、蔗糖、麦芽糖、乳糖和棉籽糖);采用NH₂P-50-4E色谱柱,以乙腈-0.1%三乙胺水溶液为流动相,建立高效液相色谱梯度洗脱,蒸发光散射法测定保健食品中8种糖。 结果 在5.0~400.0 mg/L范围内,色谱峰面积的对数与8种糖质量浓度的对数呈线性关系,最低检测质量浓度范围为5.0~20 mg/L,检测精密度<1.5%,样品加标回收率为86.7%~104.5%。 结论 液相色谱-蒸发光散射法适用于保健食品中多种糖的含量测定,该方法具有操作简单、抗干扰能力强和准确灵敏等优点。     

气相色谱-质谱法测定水中丁香酚类麻醉剂及其代谢物的研究

目的 建立水中丁香酚类麻醉剂及其主要代谢物的气相色谱-质谱检测方法。 方法 待测物通过HLB固相萃取小柱富集,经乙酸乙酯洗脱,使用气相色谱-质谱法测定。 结果 方法对丁香酚等物质的检出限可达0.01 μg/L,样品加标回收率为78.6%~104%,相对标准偏差≤7.0%(n=5)。 结论 新方法灵敏、准确,可用于环境水体中丁香酚类鱼用麻醉剂及其主要代谢物污染的检测。     

Quantitative determination of urinary hippuric acid and m- or p-Methylhippuric Acid as Indices of toluene and m- or p-Xylene exposure by high performance liquid chromatography

Summary A high performance liquid chromatographic method for the determination of hippuric and m-(p-)methylhippuric acids in urine, metabolites of toluene and m-(p-)xylene, is described. A stainless-steel column packed with octadecyl silanized silica gel was used and the mobile phase was a mixed solution of methanol/water/acetic acid (20/80/0.2). The method is simple and specific. Urine can be analyzed directly, without solvent extraction or pretreatment. The method has a lower detection limit of 0.2 g on column. All the analysis and quantitative determination can be performed within about 30 min. for samples containing m-(p-)methylhippuric acid.     

Measurement by gas chromatography of urinary hippuric acid and methylhippuric acid as indices of toluene and xylene exposure.

A gas chromatographic method was applied to the determination of the urinary glycine conjugates, hippuric, o-, m- and p-methylhippuric acids. These were extracted with ethyl acetate from urine after acidification with hydrochloric acid. The internal standard solution (heptadecanoic acid methanol solution) was added before extraction and a diazomethane-ether-ethanol solution was subsequently added to the dried extracts. The methylated residues were dissolved in methanol and injected into a gas chromatograph as described by Buchet and Lauwerys (1973). By the combined use of gas chromatography and mass spectrometry the methyl esters of hippuric acid and m-methylhippuric acid were identified in the urine of a volunteer who had been exposed to toluene and m-xylene vapours. When the urine specimen contained salicyluric acid (a urinary metabolite of salicylic acid) two sharp peaks were observed. The faster peak coincided with m- or p-methylhippuric acid. The upper limit of urinary hippuric acid concentration in healthy subjects with no occupational exposure was calculated by this method to be 1.026 microgram/ml (fiducial limit 5%) after correction to 1.024 for variation in urinary density.     

离子色谱法测定尿中甲酸的含量

目的 建立一种简单、准确的检测方法,用于甲醇中毒尿样中甲酸的快速定量分析。方法 取尿样稀释、过膜,采用万通881型离子色谱仪进行分析,淋洗液为0.5 mmol/L H₂SO₄溶液,待测物分离柱为Metrosep Organic Acids有机酸交换柱,检测器为电导检测器。结果 尿中甲酸的方法检出限为0.2 mg/L,在0.5~10.0 mg/L浓度范围内线性关系良好,相关系数为0.9997,样品加标回收率在98.2%~102.7%之间,相对标准偏差(relative standard deviation,RSD)在2.8%~4.7%之间。结论 该方法前处理简单,选择性好、准确度高、精密度好,适合尿液中甲酸的快速定量测定,可为甲醇中毒调查和治疗提供参考数据。     

Urinary trans-trans muconic acid (exposure biomarker to benzene) and hippuric acid (exposure biomarker to toluene) concentrations in Mexican women living in high-risk scenarios of air pollution

This study aimed to determine t,t-muconic acid (t,t-MA; exposure biomarker for benzene) and hippuric acid (HA; exposure biomarker for toluene) concentrations in the urine of women living in Mexico. In a cross-sectional study, apparently healthy women (n = 104) were voluntarily recruited from localities with a high risk of air pollution; t,t-MA and HA in urine were quantified using a high-performance liquid chromatography (HPLC) technique. Mean urinary levels of t,t-MA ranged from 680 to 1,310 μg/g creatinine. Mean values of HA ranged from 0.38 to 0.87 g/g creatinine. In conclusion, compared to data recently reported in literature, we found high urinary levels of t,t-MA and HA in assessed women participating in this study. We therefore deem the implementation of a strategy aimed at the reduction of exposure as a necessary measure for the evaluated communities.     

A new derivatization procedure for the analysis of hippuric acid and m-methyl-hippuric acid by gas chromatography

Summary The industrial solvents, toluene and xylene, have physicochemical properties that can be hazardous to the workers exposed. Since hippuric acid and m-methyl-hippuric acid represent the products of toluene and xylene biotransformation in urine, they are used as biological markers in studies on occupational exposure to these solvents. Several methods have been used to determine hippuric acid and m-methyl-hippuric acid —either based on gas chromatography or on high-performance liquid chromatography. In this study we propose the derivatization of hippuric acid and methyl-hippuric acid using methanol in acid medium (HCl), a low-cost reagent with a low level of toxicity. The method has been routinely used in our laboratory for 1 year and has proven to be a reliable procedure for the biological control of occupational exposure to toluene and/or xylene.     

Quantitative determination of hippuric and m-methylhippuric acids in urine by high-speed liquid chromatography.

High-speed liquid chromatography employing an ultraviolet photometric detector has been applied to the simultaneous determination of hippuric and m-methylhippuric acids in urine. Reversed-phase partition chromatography is carried out on a muBondapak C18 column with methanol-water as the eluent system. This method obviates the necessity for isolation or reaction of these acids before assay. The only pretreatment necessary is extraction of sample with ethyl acetate. A linear relationship is obtained between the peak heights and the hippuric or m-methylhippuric acid concentrations. Mean recovery of hippuric and m-methylhippuric acids in urine is 99.8% and 99.3% respectively. The determination of hippuric acid by this method gives lower concentrations in normal urine than does the colorimetric method of Umberger and Fiorese (1963).     

Quantitative determination of hippuric and m-methylhippuric acids in urine by high-speed liquid chromatography

High-speed liquid chromatography employing an ultraviolet photometric detector has been applied to the simultaneous determination of hippuric and m-methylhippuric acids in urine. Reversed-phase partition chromatography is carried out on a muBondapak C18 column with methanol-water as the eluent system. This method obviates the necessity for isolation or reaction of these acids before assay. The only pretreatment necessary is extraction of sample with ethyl acetate. A linear relationship is obtained between the peak heights and the hippuric or m-methylhippuric acid concentrations. Mean recovery of hippuric and m-methylhippuric acids in urine is 99.8% and 99.3% respectively. The determination of hippuric acid by this method gives lower concentrations in normal urine than does the colorimetric method of Umberger and Fiorese (1963).     

顶空气相色谱-质谱法同时测定尿中5种有机溶剂接触的生物监测指标

目的 建立尿中丙酮、二氯甲烷、甲乙酮、1-溴丙烷、甲基异丁基甲酮的顶空气相色谱-质谱测定方法。方法 尿样经顶空自动进样器进样,气相色谱柱分离,质谱检测器检测。结果 本方法测定尿中各组分在各浓度范围内线性良好,相关系数为0.9991～0.9996,各组分检出限分别为0.40 mg/L、0.60μg/L、0.02 mg/L、0.09μg/L、0.01 mg/L,方法精密度为1.8%～4.9%,样品加标回收率为96.9%～104.5%。结论 本方法检出限低,准确度、精密度高,样品前处理简单,适用于尿中该5种生物监测指标的同时测定。     

超高效液相色谱-质谱/质谱法测定黄酒中的甜蜜素

目的建立超高效液相色谱-质谱/质谱法测定黄酒中甜蜜素的分析方法。方法黄酒样品经0.1%甲酸-水溶解,超声提取,以0.1%甲酸水-乙腈为流动相,梯度洗脱,经Zobax C18色谱柱分离,采用MRM正负模式同时采集,外标法定量。结果该方法检出限为0.030 mg/kg;线性范围为10 ng/ml^1000 ng/ml,相关系数为0.999,在添加水平为0.1μg/kg、0.5μg/kg、2.5μg/kg时,回收率为95%~102%,RSD值为1.0%~4.5%(n=6)。结论该方法操作简便、准确、灵敏度高、稳定性好,可用于黄酒中甜蜜素的测定。     

Improved high performance liquid chromatography determination of hippuric acid and methylhippuric acid isomers in urine

A study was made on the effect of detection wavelength and separation mode of HPLC on determination of urinary hippuric acid (HA) and three isomers of methylhippuric acid (MHA). The interference of other constituents of urine in the determination was effectively decreased by detection at a short wavelength of 227.6 nm. Meta and para MHAs were separated by the addition of beta-cyclodextrine to the mobile phase. Four metabolites were successfully separated from other components of urine by the combination of ODS-silica packed-column and mobile phase (method F). The detection limits were found to be 50 and 5 mg/l for HA and MHAs, respectively. MHAs could not be detected in the non-exposed subjects. Average levels (+/- SD) of HA in non-exposed males and females were 272.2 (+/- 210.8) and 393.0 (+/- 269.8) mg/l, respectively. The urinary levels of HA in females were significantly higher than those in males.