Quantitation of urinary o-xylene metabolites of rats and human beings by high performance liquid chromatography

Summary A high performance liquid chromatographic method for the determination of urinary o-xylene metabolites of rats, volunteers, and workers was described. In rat urine, the major metabolite, indicated by the glucuronic acid reaction, was separated with thin-layer liquid chromatography and identified as o-toluic acid glucuronide by high performance liquid chromatography. Another rather minor metabolite was demonstrated to be o-methyl hippuric acid. One of the major urinary metabolite in volunteers administered o-xylene orally was demonstrated to be o-methylhippuric acid and another minor metabolite was o-toluic acid glucuronide. In the urine of volunteers exposed to 138 ppm of o-xylene for 3 h in an artificial exposure chamber, o-methylhippuric acid was found to be the major metabolite and a minute amount of o-toluic acid glucuronide was found to be the minor metabolite. In the urine of shipbuilding workers, using a thinner containing toluene and xylenes (o-, m- and p-), hippuric acid and methylhippuric acids (o-, m- and p-) were recognized. Thus, urinary o-methylhippuric acid could be an index of o-xylene exposure of workers.     

尿2－硫代噻唑烷－4－羧酸的反相高效液相色谱法研究

应用反相高效液相色谱法（分离柱：ＵｔｒａｓｐｈｅｒｅＴＭＯＤＳＣ－１８）对二硫化碳（ＣＳ２）代谢产物２－硫代噻唑烷－４－羧酸（ＴＴＣＡ）进行了测定条件优选。以流动相为甲醇（１４％）、冰乙酸（１. ５％）和水（８４. ５％）；流速１. ５ｍｌ／ｍｉｎ；紫外线检测波长２７２ｎｍ时分离检测效果最好。同时，对尿样前处理方法也进行了探讨。本法平均回收率９８. ９％，最小检出量０. ００８ｎｇ，ＴＴＣＡ浓度在０～２００ｍｇ／Ｌ范围内呈直线相关（ｒ＝０. ９９９６）是ＣＳ２生物监测的良好方法。     

尿2－硫代噻唑烷－4－羧酸的反相高效液相色谱法研究

应用反相高效液相色谱法（分离柱：ＵｔｒａｓｐｈｅｒｅＴＭＯＤＳＣ－１８）对二硫化碳（ＣＳ２）代谢产物２－硫代噻唑烷－４－羧酸（ＴＴＣＡ）进行了测定条件优选。以流动相为甲醇（１４％）、冰乙酸（１. ５％）和水（８４. ５％）；流速１. ５ｍｌ／ｍｉｎ；紫外线检测波长２７２ｎｍ时分离检测效果最好。同时，对尿样前处理方法也进行了探讨。本法平均回收率９８. ９％，最小检出量０. ００８ｎｇ，ＴＴＣＡ浓度在０～２００ｍｇ／Ｌ范围内呈直线相关（ｒ＝０. ９９９６）是ＣＳ２生物监测的良好方法。     

Quantitation of urinary metabolites of toluene,xylene, styrene,ethylbenzene, benzene and phenol by automated high performance liquid chromatography

Summary An automated high performance liquid chromatographic method (HPLC) for the direct determination of urinary concentrations of hippuric acid (HA), and o-, m- and p-methyl hippuric acids (MHAs), metabolites of toluene and o-, m-, and p-xylenes, and of urinary phenyl glyoxylic acid (PGA) and mandelic acid (MA), metabolites of styrene or ethylbenzene, is described. Methanol was added to urine, the mixture was centrifuged and the supernatant was injected into HPLC. A stainless-steel column packed with octadecyl silanized silicate was used and the mobile phase was a mixed solution of 5 mM potassium phosphate monobasic/acetonitrile (90/10). The method is simple and specific. Urine can be analyzed without solvent extraction. Analysis can be performed satisfactorily within 45 min for samples containing HA, MHAs, PGA and MA, and within 15 min for those containing HA, PGA and MA. Another automated HPLC method for the determination of urinary concentrations of phenylsulfate (PhS) and phenylglucuronide (PhG), metabolites of benzene and phenol, is also described. Urine was centrifuged and the supernatant was injected into HPLC. A column packed with octadecyl silicate and a mobile phase of 50 mM of potassium phosphate monobasic/acetonitrile (85/15) were used. The whole analyses and quantitative determination can be performed within 15 min for samples containing PhS and PhG in the worker's urine with a simple mobile phase. The accuracy and precision in the present methods by the use of automated HPLC were satisfactory.     

Determination of picloram in soil and water by reversed-phase liquid chromatography

A reversed-phase liquid Chromatographic method is presented for the determination of picloram in the parts per billion 2 (ppb) range in soil, soil solution, and stream samples. Quantification is effected by UV absorption at 254 nm. Derivatization is not necessary. The method permits 92% ±7.1 recovery from water samples and 61.8% ± 11.1 recovery from soil samples.     

Application of high-pressure liquid chromatography in the analysis of urinary metabolites of aromatic solvents

The introduction of HPLC methods in industrial toxicology represents a valuable tool for the routine monitoring of workers occupationally exposed to aromatic solvents. The HPLC method described here permits the simultaneous determination of metabolites of ethylbenzene, styrene, toluene, xylene isomers, benzene, phenol and cresol isomers in diluted urine samples. Pretreatment of urine samples with steam distillation is necessary only for determination of phenol and cresols because of their low concentrations. A comparison between a GLC and the HPLC procedure for mandelic and phenylglyoxylic acids confirmed the satisfactory performance of the HPLC method.     

Biochemical markers for assessment of niacin status in young men: urinary and blood levels of niacin metabolites

Biochemical markers of niacin status were studied in healthy young men fed 6.1 to 32 niacin equivalents (NE) per day over an 11-wk period while residing in a metabolic unit. Methylated metabolites of niacin, N1-methylnicotinamide (NMN) and N1-methyl-2-pyridone-5-carboxamide (2-pyr), in urine and plasma were determined during periods of low (6.1 or 10.1 NE per day), adequate (19 NE per day = 1 RDA) and high (25 or 32 NE per day) niacin intakes and after small test doses of nicotinamide. Urine excretion of less than 1.2 mg/d of either NMN or 2-pyr was a reliable indicator of subjects receiving the lowest intake of 6.1 NE/d, but the NMN metabolite was a better marker of subjects ingesting 10.1 NE/d. The ratio of 2-pyr/NMN in urine was not as good a measure of the 6.1 NE/d intake as the individual metabolite excretions and was not responsive to the 10.1 NE/d intake. Plasma niacin metabolites were generally not as reliable as urinary metabolites for identifying subjects receiving low niacin intakes, however, values for plasma 2-pyr dropped quickly and were eventually nondetectable. After a 1 RDA oral dose of nicotinamide, increases in urine and plasma 2-pyr levels above pre-dose baseline values were significantly decreased in subjects receiving low, as compared to adequate, niacin intake. A leucine supplement had no effect on the rate of repletion of niacin-deficient subjects nor on the level of methylated niacin metabolites in urine or plasma.     

人血浆中头孢氨苄浓度高效液相色谱法测定

目的建立高效液相色谱法测定人血浆中头孢氨苄的浓度。方法采用噻克硝唑为内标,色谱柱为YMC ODS-A C18(150 mm×4.6 mm),流动相为乙腈∶甲醇∶pH 4.5的醋酸盐缓冲液(6∶10∶84),流速为1.0 mL/min,检测波长为260 nm。结果头孢氨苄血清浓度在0.101~32.100μg/mL范围内具有良好线性关系,最低定量限为0.101μg/mL,方法回收率为97.5%~103.0%,日内、日间相对标准差均<7%。结论高效液相色谱法简便、准确、灵敏,适用于头孢氨苄血药浓度监测和药代动力学研究。     

果酒中11种酚类化合物反相高效液相色谱检测方法的建立

目的采用反相高效液相色谱法(RP-HPLC)建立11种酚类化合物的检测方法,同时测定其在果酒中的含量。方法色谱柱:Agilent Eclipse XDB C18(4.6mm×250mm,5μm);流动相:梯度洗脱,A相:0.1%磷酸;B相:甲醇,洗脱程序:10%B等度(0～10min);10%～20%B线性变换(10～15min);20%B等度(15～25min);20%～60%B线性变换(25～65min);60%B等度(65～75min);检测波长:210、270、320、327、360nm;柱温30℃;流速1.0ml/min。结果在10～500μg/ml浓度范围内,没食子酸、绿原酸、表儿茶素、阿魏酸、白皮杉醇、金丝桃苷、木犀草素7种功能成分均具有良好的线性相关(R2均≥0.9995);在8～400μg/ml浓度范围内,牡荆素、白藜芦醇、槲皮素、山奈酚4种功能成分均具有良好的线性相关(R2=1.0000);回收率为85.1%～112.0%,RSD均<2%;11种功能成分的定量限为0.32～1.59ng,检测限为0.10～0.48ng。结论建立的方法准确、可靠,实现了11种功能成分的分离分析,可用于其在不同种类果酒中的含量测定。     

Quantitative determination of urinary lipid metabolites by high pressure liquid chromatography as indicators of menadione-induced in vivo lipid peroxidation

The one- and two-electron-reduction reactions of menadione result in the generation of reactive oxygen species which are believed to mediate the cytotoxicity of this xenobiotic. The induction of lipid peroxidation in liver and isolated hepatocytes occurs in response to the menadione-mediated formation of reactive oxygen species. However, studies on the effects of menadione on the urinary excretion of lipid metabolites have not been conducted. The effect of a single oral dose of 60 mg menadione/kg to rats on the urinary excretion of the lipid metabolites malondialdehyde (MDA), formaldehyde (FA), acetaldehyde (ACT), and acetone (ACON) has been examined over 48 h post-treatment. The urinary metabolites were identified by gas chromatography-mass spectrometry and quantitated by high pressure liquid chromatography. Time-dependent increases in the urinary excretion of the four metabolites were observed after menadione administration. Over the 48 h of the study, the menadione-induced urinary excretion of MDA, FA, ACT, and ACON increased by approximately 1.5-, 2.0-, 1.7-, and 3.2-fold, respectively, relative to control animals. The data were expressed in nmoles/kg body weight/4.5 h. The results clearly demonstrate that menadione increases the urinary excretion of four lipid metabolites. These metabolites may have widespread applicability as biomarkers of altered lipid metabolism in disease states and exposure to environmental pollutants/xenobiotics which induce enhanced lipid peroxidation. The non-invasive methods offer advantages over most other methods for assessing oxidative stress in vivo.     

反相高效液相色谱法同时测定化妆品中的7种性激素

目的：建立一种简单、准确、灵敏、同时测定化妆品中7种性激素（雌二醇、雌三醇、雌酮、睾酮、甲基睾酮、孕酮、己烯雌酚）的高效液相色谱检测方法。方法：样品经甲醇提取、净化后，在XTerra RP18色谱柱（5μm，4．6mm&#215;150mm）上，以桕甲醇／乙腈（50：30：20，v／v／v）为流动相，利用二极管阵列检测器进行检测，雌二醇、雌三醇、雌酮、己烯雌酚的检测波长为197nm，睾酮、甲基睾酮、孕酮的检测波长为240nm。结果：7种性激素分离良好并排除了样品中杂质峰的干扰。样品的平均回收率范围为92．0％～95．2％；相对标准偏差为1．60％～2．65％；检出限为0．15～1．0mg／L。结论：该方法简便、准确并可同时测定化妆品中的7种性激素。     

湿膜萃取-反向高效液相色谱法测定水中环境类激素

建立了湿膜萃取 -梯度洗脱反向高效液相色谱同时测定 7种环境类激素的方法 ,优化了色谱条件和湿膜萃取条件 ,并应用于实际水体的测定。     

离子对高效液相色谱法分析米曲霉发酵液中的曲酸

为建立米曲霉培养液中曲酸的离子对高效液相色谱法 ,采用 0 0 1mol L磷酸氢二钠及 2mmol四丁基溴化胺做流动相 ,选择 2 2 6nm紫外吸收波长 ,测定米曲酶发酵液中的曲酸 ,曲酸在C1 8色谱柱上获得良好的分离。本方法检出限为 0 0 1 2 μg ml;麦芽汁 蛋白胨、麦芽汁 酵母膏、马铃薯 酵母膏 蔗糖三种培养液中回收率分别为 98 2 %、1 0 3 4%、97 2 % ;RSD分别为 0 51 %、0 45 %、0 43 %。     

反相高效液相色谱法同时测定广州部分地区肉类4种抗生素的残留量

〔目的〕高效液相色谱法同时测定肉类中磺胺嘧啶 (SD)、磺胺二甲基嘧啶 (SMZ)、土霉素 (OTC)、磺胺甲恶唑(SN2 )抗生素的残留量。〔方法〕色谱柱为ODS -C1 8,以 2mmol L醋酸铵 :乙腈 :80 :2 0 (pH =2 .0 3 )为流动相 ,流速 0 .8ml/min ,486紫外检测器 ,检测波长为 2 70nm。〔结果〕该方法各抗生素的线性范围均为 0 .0 1～ 1.0 μg/ml ,相关系数均为 0 .99以上 (n =5 ) ;最低检测限为 0 .0 1μg/ml;回收率均为 95 .6%～ 10 6.0 % ;日内、日间变异系数均在允许范围内 (小于 15 % )。〔结论〕该方法操作简便 ,是动物性食品残留抗生素的快速而准确的检测方法。