高效液相色谱与液质联用测定尿中苯巯基尿酸的方法比较
Determination of Urinary S-phenylmercapturic Acid by HPLC and LC/MS: A Comparison of the Two Methods
-
摘要:
目的 通过高效液相色谱(HPLC)与液质联用(LC/MS)检测尿中苯巯基尿酸(SPMA)的方法比对,评价HPLC方法的可行性。 方法 尿样经50%硫酸酸化、氯仿 : 异丙醇(体积比为5 : 1)萃取,挥干残渣用甲醇溶解后经十八烷基硅烷(ODS)柱分离,分别采用紫外检测器和质谱检测器检测;对86 名职业苯接触者尿样采用两种方法比对分析,以LC/MS 分析数据评定HPLC 测定结果的有效性。 结果 HPLC 方法和LC/MS 方法对SPMA的检出限(LOD)分别为40 μg/L 和10 μg/L,SPMA保留时间分别为31 min 和7.8 min 左右。86 份尿样中有13 份为假阳性,两方法检测结果的相关系数为0.915,但HPLC检测数据高估LC/MS数据8%~21%。 结论 HPLC法可作为职业苯接触者生物监测的初步筛检方法,LC/MS 可准确定量测定尿中SPMA浓度。 Abstract:Objective To compare the validity of HPLC and LC/MS analysis of urinary S-phenylmercapturic acid (SPMA)so as to assess the feasibility of HPLC method in measuring SPMA. Methods The urine samples were acidified by 50% sulfuric acid and pretreated by liquid-liquid extraction using chloroform : isopropyl alcohol(5 : 1, v/v). The residue dissolved with methanol was injected in ODS-3 column for SPMA separation, followed by ultraviolet detector and LC/ESI-NI/ MS analysis, respectively. Paired measurements were applied to the analysis of 86 urine samples from workers occupationally exposed to benzene. HPLC results were evaluated by comparison with the LC/MS analysis. Results The limits of detection of HPLC and LC/MS were 40 μg/L and 5 μg/L, respectively. Retention time of SPMA peak appear at about 31 min and 7.8 min, respectively. 13 of 86 samples in HPLC analysis were assigned as false-positives. HPLC results compared with LC/MS data showed to overestimate SPMA concentrations of about 8%-21% and a correlation coefficient of 0.915 was calculated between the two methods. Conclusion HPLC analytical method is suitable for preliminary screening of urinary SPMA in routine biomonitoring for benzene-exposed workers, while LC/MS could be an more accurate quantitative determination method for measuring urinary SPMA concentration.
下载:
