Rapid screening of inorganic poisons by column-switching ion chromatography

柱切换离子色谱法快速筛查无机毒物

• 批准号: 13557037
• 负责人: ISHII Akira
• 金额: $ 1.79万
• 依托单位: Fujita Health University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557037/
• 关键词: Ion chromatography; Sodium azide; Semimicrocolumn; Quantitaion; Conway diffusion method; 亜ヒ酸

In 1998, an extraordinary murder incidence took place in Wakayama ; 4 victims were killed. After the incidence, a lot of imitative poisoning cases occurred. Several inorganic compounds, such as arsenic, cyanide or azides, were used in the poisoning cases. In the case of sodium azide, it was then regulated as a poison by the Poisonous and Deleterious Substances Control Law in 2000. It is thus important to screen these inorganic compounds rapidly with less labor. Ion chromatography (IC) has been widely used for the separation of inorganic cations and anions ; IC is easy to use and highly quantitative, although IC cannot identify and specify these inorganic ions. So far, IC has been applied mainly for environmental samples, such as surface water samples. In this project, we have tried to develop simple, rapid and robust methods for the analyses of inorganic ions in the biological samples by IC.Sodium azide is widely used as preservative for laboratory reagents. However, it is highly toxic … More and the intake of about I g sodium azide would be fatal. In rat, its oral LD_<50> value is estimated to be 45 mg/kg, which is only 5 times greater than that potassium cyanide. Thus we have developed a simple method for analysis of sodium azide. We tried several methods for purification of azide from biological samples ; it was revealed that Conway diffusion method would be simple and fast. Azide ion in various samples was extracted using a Conway microdiffusion cell ; hydrazoic acid was vaporized from 1 mL sample (plasma or beverages) by adding 1 mL of 5% sulfuric acid in the outer groove, and absorbed in 250 μL of 0.1 M NaOH solution in the central round basin. Incubation for 30 min at 37℃ was sufficient for the purification of azide from biological samples. For IC analysis, a 20-μL aliquot was subjected to IC system equipped with a suppressor and a conductivity detector (Dionex, DX 500 system). The precolumn and separation column used were an AG 15 (20 mm × 2 mm i.d.) and an AS 15 (250 mm × 2 mm i.d.) semimicrocolumns. The mobile phase was 38 mM NaOH and the flow rate was set at 0.4 mL/min. The retention time of azide ion was about 10 mm. Combination of the reduction of NaOH solution in the Conway diffusion method and the use of semimicrocolums, has enabled 4 to 5 times higher sensitivity compared to methods previously reported ; the detection limit for serum samples was about 30 ng/mL Its calibration curve showed good linearity in the range of 50 ng/mL and 10 μg/mL serum We have validated this method; the CV values for intra-day and day-to-day measurements were less than 6.8 % and 93 %, respectively. In the case of beverages, Comway diffusion method was needed for accurate quantitaion because of the interference of some impurity peaks. Azide ion was detected from rat serum samples 1 or 4 hours after its oral administration (10 mg/kg). Thus the present method can be applicable for clinical and forensic toxicology, because of its simplicity and sensitivity. We are now preparing a paper for submitting. In the next research, we are going to obtain data about the decay of azide ion of different samples under several environmental conditions. Less

1998年，和歌山发生了一起非同寻常的杀人事件； 4名受害者被杀。事件发生后，多起模仿中毒事件发生。中毒案件中使用了几种无机化合物，例如砷、氰化物或叠氮化物。就叠氮化钠而言，2000年《有毒有害物质管理法》将其列为毒物。因此，以较少的劳动力快速筛选这些无机化合物非常重要。离子色谱（IC）已广泛用于无机阳离子和阴离子的分离；尽管 IC 无法识别和指定这些无机离子，但 IC 易于使用且高度定量。迄今为止，IC主要应用于环境样品，例如地表水样品。在这个项目中，我们试图开发简单、快速和稳健的方法，通过离子色谱法分析生物样品中的无机离子。叠氮化钠广泛用作实验室试剂的防腐剂。但它具有剧毒，摄入约1克叠氮化钠就会致命。在大鼠中，其口服LD_50值估计为45mg/kg，仅比氰化钾高5倍。因此，我们开发了一种简单的叠氮化钠分析方法。我们尝试了多种从生物样品中纯化叠氮化物的方法；结果表明康威扩散法简单且快速。使用康威微扩散池提取各种样品中的叠氮化物离子；在外槽中加入 1 mL 5%硫酸，从 1 mL 样品（血浆或饮料）中蒸发出叠氮酸，并在中心圆盆中吸收到 250 μL 0.1 M NaOH 溶液中。 37℃孵育30分钟足以从生物样品中纯化叠氮化物。对于 IC 分析，将 20 μL 等分试样置于配备抑制器和电导检测器的 IC 系统（Dionex，DX 500 系统）中。使用的预柱和分离柱是 AG 15 (20 mm × 2 mm i.d.) 和 AS 15 (250 mm × 2 mm i.d.) 半微柱。流动相为 38 mM NaOH，流速设置为 0.4 mL/min。叠氮化物离子的保留时间约为10mm。康威扩散法中 NaOH 溶液的还原和半微柱的使用相结合，与之前报道的方法相比，灵敏度提高了 4 至 5 倍；血清样品的检出限约为30 ng/mL，其校准曲线在50 ng/mL和10 μg/mL血清范围内表现出良好的线性，我们对该方法进行了验证；日内和日常测量的 CV 值分别小于 6.8% 和 93%。对于饮料，由于一些杂质峰的干扰，需要使用Comway扩散法来准确定量。口服给药（10mg/kg）后1或4小时从大鼠血清样品中检测到叠氮离子。因此，由于其简单和灵敏，本方法可适用于临床和法医毒理学。我们现在正在准备提交论文。在下一步的研究中，我们将获得不同样品在多种环境条件下叠氮离子衰变的数据。较少的

项目成果

T.Sano: "Sensitive determination of midazolam and identification of its two metabolites in human body fluids by column-switching capillary high-performance liquid chromatography/fast atom bombardment-mass spectrometry."Legal Medicine. 3. 149-156 (2001)

T.Sano：“通过柱切换毛细管高效液相色谱/快原子轰击质谱法灵敏测定人体液中的咪达唑仑并鉴定其两种代谢物。”法律医学。

津田孝雄: "カフェイン錠剤使用後のヒト汗中カフェイン及びその代謝物の存在確認とそれらの濃度経時変化"発汗学. 8. 13-19 (2001)

Takao Tsuda：“使用咖啡因片后人体汗液中咖啡因及其代谢物的存在及其浓度随时间的变化的确认”Shidrology. 8. 13-19 (2001)。

Y.Mizuno: "Identification and characterization of 17 phenothiazine compounds by capillary high-performance liquid chromatography/fast atom bombardment mass spectrometry."Legal Medicine. 4. 207-216 (2002)

Y.Mizuno：“通过毛细管高效液相色谱/快原子轰击质谱法鉴定和表征 17 种吩噻嗪化合物。”法律医学。

Hamajima M.et al.: "Simple analysis of acetaminophen in human plasma by solid-phase micro-extraction and gas chromatography."Jpn J Forensic Toxicol. 22. 22-26 (2004)

Hamajima M.等人：“通过固相微萃取和气相色谱法简单分析人血浆中的对乙酰氨基酚。”Jpn J Forensic Toxicol。

Kurihara R, Ishii A, Watanabe-Suzuki K, Kamazawa T, Seno H, Suzuki O, Katsumata Y.: "Simple extraction of gamma-hydroxybutyrate in human whole blood by headspace solid-phase microextraction."Jpn J Forensic Toxicol. 19. 38-45 (2001)

Kurihara R, Ishii A, Watanabe-Suzuki K, Kamazawa T, Seno H, Suzuki O, Katsumata Y.：“通过顶空固相微萃取简单提取人全血中的 γ-羟基丁酸盐。”Jpn J Forensic Toxicol。