叶海朋, 邵吉, 谈思维, 单晓月, 张玲, 傅红, 张磊. 工作场所空气中双乙酰的4-硝基邻苯二胺柱前衍生-高效液相色谱测定方法[J]. 环境与职业医学, 2023, 40(9): 1079-1084. DOI: 10.11836/JEOM23073
引用本文: 叶海朋, 邵吉, 谈思维, 单晓月, 张玲, 傅红, 张磊. 工作场所空气中双乙酰的4-硝基邻苯二胺柱前衍生-高效液相色谱测定方法[J]. 环境与职业医学, 2023, 40(9): 1079-1084. DOI: 10.11836/JEOM23073
YE Haipeng, SHAO Ji, TAN Siwei, SHAN Xiaoyue, ZHANG Ling, FU Hong, ZHANG Lei. Determination of diacetyl in workplace air by high performance liquid chromatography using 4-nitro-o-phenylenediamine as precolumn derivatization[J]. Journal of Environmental and Occupational Medicine, 2023, 40(9): 1079-1084. DOI: 10.11836/JEOM23073
Citation: YE Haipeng, SHAO Ji, TAN Siwei, SHAN Xiaoyue, ZHANG Ling, FU Hong, ZHANG Lei. Determination of diacetyl in workplace air by high performance liquid chromatography using 4-nitro-o-phenylenediamine as precolumn derivatization[J]. Journal of Environmental and Occupational Medicine, 2023, 40(9): 1079-1084. DOI: 10.11836/JEOM23073

工作场所空气中双乙酰的4-硝基邻苯二胺柱前衍生-高效液相色谱测定方法

Determination of diacetyl in workplace air by high performance liquid chromatography using 4-nitro-o-phenylenediamine as precolumn derivatization

  • 摘要:
    背景  双乙酰(DC)被广泛应用于食品调味行业,过量职业接触会引起人体严重的呼吸系统疾病,但目前对于工作场所空气中DC尚缺乏相应的国家标准检测方法。
    目的  建立4-硝基邻苯二胺(NPDA)柱前衍生-高效液相色谱法测定工作场所空气中DC的方法。
    方法  用溶液吸收法采集工作场所空气中DC,采用4-硝基邻二苯胺作为衍生化试剂。通过调整吸收液磷酸浓度,优化反应试剂比例、反应温度和时间,建立测定工作场所空气中DC的高效液相色谱方法,得到线性、检出限、定量下限等性能指标。采用相对比较法评价方法采样效率,吸收液保存试验评价样品稳定性,加标回收试验评价方法准确度和精密度,并进行干扰试验。将所建立的方法应用于实际样品检测,考察方法的适应性。
    结果  衍生化反应温度越高,反应时间越长,衍生化效率越高,因此选择60 ℃衍生化2 h。反应液经SB-C18色谱柱(250 mm×4.6 mm,5 μm)分离,在30 ℃柱温下,用甲醇-水(体积比:65%/35%)混合液为流动相,以1.0 mL·min−1的流速洗脱,紫外检测器(λmax=257 nm)检测,保留时间定性,外标法定量。本法DC检测范围为5~2000 μg·L−1,相关系数为0.999 9,检出限为1.3 μg·L−1,定量下限为4.3 μg·L−1,最低检出浓度为4.3 μg·m−3,最低定量浓度为14.3 μg·m−3(以采样体积V0=3.0 L计)。本法样品加标回收率为99.1%~100.8%,批内精密度为0.5%~3.0%,批间精密度为1.2%~2.0%。本法平均采样效率为94.5%,样品在4 ℃下至少可保存14 d,工作场所空气共存成分不干扰DC的测定。本法检测某香精生产车间空气,DC含量为5.86~8.85 mg·m−3
    结论  采用1.0%磷酸吸收液采集、NPDA柱前衍生-高效液相色谱法测定空气中DC,方法简单实用,准确度好,灵敏度高,DC吸收液无损失降解,可应用于工作场所空气中DC的检测。

     

    Abstract:
    Background  Diacetyl (DC) is widely used in the food flavoring industry and excessive occupational exposure to DC can cause serious respiratory diseases. However, there is no corresponding national standard method for the determination of DC in the air of workplace.
    Objective  To establish a method for the determination of DC in workplace air by high performance liquid chromatography using 4-nitro-o-phenylenediamine (NPDA) as precolumn derivatization.
    Methods  DC in the air of workplace was collected by solution absorption method. This experiment used NPDA as the derivatization reagent. By adjusting acidity of solution and optimizing concentration ration of DC/NPDA, derivatization temperature, and time, a method for the determination of DC in workplace air was proposed, and its performance indexes such as linearity, detection limit, and lower limit of quantification were obtained. Sampling efficiency was evaluated by relative comparison method, and sample stability was evaluated by sample preservation test. Accuracy and precision of the method were evaluated by standard addition recovery test with blank samples, and an interference test was carried out by adding standard samples. The established method was applied to actual samples to evaluate its adaptability.
    Results  A combination of 60 °C for 2 h was selected for derivatization because a higher derivatization reaction temperature and a longer reaction time associated with a higher derivatization efficiency. The solution was separated by SB-C18 column (250 mm×4.6 mm, 5 μm) at 30 ℃, using a mixture of methanol and water (v/v, 65%/35%) as mobile phase with an elution flow rate of 1.0 mL·min−1, and was detected with a variable wavelength detector (λmax=257 nm) by qualitative analysis based on retention time and quantitative analysis based on external standard method. In terms of the proposed method, the linear range of detection was from 5 μg·L−1 to 2000 μg·L−1, with a correlation coefficient of 0.9999, and a detection limit of 1.3 μg·L−1, the quantitative detection of the lower limit was 4.3 μg·L−1, with a sampling volume V0 of 3.0 L, the minimum detection concentration was 4.3 μg·m−3, and the minimum quantitative concentration was 14.3 μg·m−3. The recovery rate was 99.1%-100.8%, the intra-batch precision was 0.5%-3.0%, and the inter-batch precision was 1.2%-2.0%. The average sampling efficiency of this method was 94.5%, and the sample could be stored at 4 °C for at least 14 d. The coexisting components in the air of the workplace did not interfere with the determination of DC. The DC content in the air of a flavor workplace was 5.86-8.85 mg·m−3.
    Conclusion  A determination method for DC in workplace air by high performance liquid chromatography using NPDA as precolumn derivatization after being collected by 1.0% phosphoric acid absorbent is proposed and has the advantages of simple operation, high sensitivity, and good accuracy. With no DC loss and degradation, the method may satisfy the request for DC determination in the air of workplace.

     

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