Mandal Debsmita, Dey Indrayani, Ghosh Chiranjit
Department of Biotechnology, Manipal Institute of Technology, Manipal Academy of Higher Education Manipal Karnataka 576104 India
Harvard Medical School 25 Shattuck Street Boston 02115 MA USA.
RSC Adv. 2024 Oct 11;14(44):32230-32238. doi: 10.1039/d4ra05907g. eCollection 2024 Oct 9.
Diabetes ketoacidosis (DKA) is a life-threatening complication and requires immediate medical attention in the case of diabetes subjects, especially in the case of type 1 diabetes mellitus. In the condition of DKA, the body produces an excess amount of ketone bodies after unregulated fat degradation, causing blood to become acidic and hampering the regular metabolic activities of the body. The current diagnostic technique for DKA condition is based on monitoring ketone bodies, especially β-hydroxybutyric acid, from human urine and blood samples. The detection of serum ketone bodies in pathology is sometimes limited due to false positive results and the lack of standardization for precise quantification of analytes. In this study, a paper-based patch operating on the thin film solid-phase microextraction (TF-SPME) principle was developed and it was coupled with gas chromatography-mass spectrometry for simple quantification of β-hydroxybutyric acid (BHB) ketone body from a phosphate-buffered saline matrix. To fabricate the paper-based TF-SPME patches, a regular A4 sheet paper sheet was utilized as the substrate and uniform coating by multiwalled carbon nanotubes (MWCNT), polydimethylsiloxane (PDMS) and divinyl benzene (DVB) compounds was performed with an automatic film applicator. The 70 μm paper-based coated sheet was trimmed into 4 cm × 1 cm dimension pieces to obtain multiple patches from a single sheet. Extraction of the BHB ketone body into the closed vials was performed by exploiting the individual DVB/PDMS and DVB/CNT/PDMS paper patches followed by desorption with acetonitrile before quantification by gas chromatography-mass spectrometry analysis. Our study showed that the BHB extraction efficiency of DVB/PDMS-coated patches was higher than that of DVB/CNT/PDMS. The outcome showed a good linearity ( = 0.99) within the 500-20 000 ng mL concentration range of BHB by paper-based DVB/PDMS patches. This study demonstrated the feasibility of utilizing simple, cost-effective paper-based disposable TF-SPME patches as a sampling kit for future screening of diabetes ketoacidosis without the need for prolonged traditional sample preparation in pathology.
糖尿病酮症酸中毒（DKA）是一种危及生命的并发症，糖尿病患者，尤其是1型糖尿病患者一旦出现这种情况，需要立即就医。在DKA状态下，身体在脂肪降解失控后会产生过量的酮体，导致血液变酸并阻碍身体的正常代谢活动。目前用于诊断DKA状态的技术是基于监测人体尿液和血液样本中的酮体，特别是β-羟基丁酸。由于假阳性结果以及缺乏用于精确分析物定量的标准化，病理检查中血清酮体的检测有时会受到限制。在本研究中，开发了一种基于薄膜固相微萃取（TF-SPME）原理的纸质贴片，并将其与气相色谱-质谱联用，用于从磷酸盐缓冲盐基质中简单定量β-羟基丁酸（BHB）酮体。为了制备基于纸质的TF-SPME贴片，使用普通的A4纸张作为基材，并通过自动涂膜器用多壁碳纳米管（MWCNT）、聚二甲基硅氧烷（PDMS）和二乙烯基苯（DVB）化合物进行均匀涂覆。将70μm的纸质涂覆片材裁剪成4cm×1cm尺寸的片，以便从单张片材中获得多个贴片。通过使用单独的DVB/PDMS和DVB/CNT/PDMS纸质贴片将BHB酮体萃取到封闭的小瓶中，然后在通过气相色谱-质谱分析进行定量之前用乙腈解吸。我们的研究表明，DVB/PDMS涂覆贴片的BHB萃取效率高于DVB/CNT/PDMS。结果表明，基于纸质的DVB/PDMS贴片在500-20000 ng/mL的BHB浓度范围内具有良好的线性（ = 0.99）。本研究证明了使用简单、经济高效的一次性纸质TF-SPME贴片作为采样试剂盒用于未来糖尿病酮症酸中毒筛查的可行性，而无需在病理检查中进行长时间的传统样品制备。
