Flow Innovation-Research for Science and Technology Laboratories (Firstlabs), Thailand; Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, Thailand.
Flow Innovation-Research for Science and Technology Laboratories (Firstlabs), Thailand; Department of Chemistry, Faculty of Science, Srinakharinwirot University, Sukhumvit 23, Bangkok, Thailand.
Anal Chim Acta. 2020 Mar 22;1103:115-121. doi: 10.1016/j.aca.2019.12.043. Epub 2019 Dec 18.
This work presents a novel development that exploits the concept of in-situ gas-separation together with a specific enzymatic colorimetric detection to produce a portable biosensor called "Blood Alcohol Micro-pad" for direct quantitation of ethanol in whole blood. The thin square device (25 mm × 25 mm × 1.8 mm) comprises two layers of patterned filter paper held together with a double-sided mounting tape with an 8-mm circular hole (the headspace). In operation, the reagent is deposited on one layer and covered with sticky tape. Then 8 μL of a blood sample is dispensed onto the opposite layer and covered with sticky tape. Diffusion of ethanol across the 1.6 mm narrow headspace permits selective detection of ethanol by the enzymatic reagents deposited on the opposite layer. This reagent zone contains alcohol oxidase, horseradish peroxidase and 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, as the chromogenic reagent. The color intensity, measured from the recorded digital image, resulting from the enzymatic assay of ethanol, correlates with the concentration of blood alcohol. The results obtained with spiked mice and sheep blood samples, using an external calibration in the range of 1-120 mg dLethanol, gave recoveries of 93.2-104.4% (n = 12). The "Blood Alcohol Micro-pad" gave good precision with %RSD <1 (50 mg dL ethanol, n = 10) and limit of quantification (10SD of intercept/slope) of 11.56 mg dL. The method was successfully validated against a headspace gas chromatography-mass spectrometric method. It has good potential for development as a simple and convenient blood alcohol sensor for on-site testing.
这项工作提出了一种新的发展，利用原位气体分离的概念和特定的酶比色检测，生产一种便携式生物传感器，称为“血液酒精微垫”，用于直接定量全血中的乙醇。这个薄的方形装置（25 毫米×25 毫米×1.8 毫米）由两层图案化滤纸组成，用双面安装胶带固定在一起，中间有一个 8 毫米的圆形孔（顶空）。在操作中，试剂被沉积在一层上并用胶带覆盖。然后将 8μL 的血液样本分配到对面的层上并用胶带覆盖。乙醇通过 1.6 毫米窄顶空的扩散允许在对面层上沉积的酶试剂选择性地检测乙醇。这个试剂区含有醇氧化酶、辣根过氧化物酶和 2'-偶氮-双（3-乙基苯并噻唑啉-6-磺酸）二铵盐，作为显色试剂。通过对乙醇进行酶测定得出的数字图像记录的颜色强度与血液酒精浓度相关。使用外部校准范围为 1-120mg dL 乙醇，对添加了老鼠和绵羊血液样本进行的测试，得到的回收率为 93.2-104.4%（n=12）。“血液酒精微垫”在 50mg dL 乙醇（n=10）时，具有小于 1%的良好精密度（%RSD），定量下限（截距/斜率的 10 倍标准差）为 11.56mg dL。该方法已成功针对顶空气相色谱-质谱法进行了验证。它具有作为简单方便的现场测试血液酒精传感器的良好发展潜力。
