Development of a Novel Method to Quantify Quinolinic Acid in Biological Samples.

IF 4.1 Q3 NEUROSCIENCES

International Journal of Tryptophan Research Pub Date : 2025-10-31 eCollection Date: 2025-01-01 DOI:10.1177/11786469251390415

Masatsuna Tasaka, Hidetsugu Fujigaki, Sayaka Sugiura, Suwako Fujigaki, Akihiro Ikuno, Yasuko Yamamoto, Masao Takemura, Akio Kimura, Kuniaki Saito

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Abstract

Background: The accumulation of quinolinic acid (QUIN) in cerebrospinal fluid and serum may be used as a biomarker for various neuropsychiatric and inflammatory diseases. In this study, we developed a highly sensitive method to measure QUIN.

Methods: A reverse-phase high-performance liquid chromatography (HPLC) with fluorescence detection was established based on the enzymatic conversion of QUIN to nicotinic acid mononucleotide by recombinant quinolinic acid phosphoribosyltransferase, followed by the formation of fluorescent (BODIPY)-labeled deamido-NAD by recombinant nicotinic acid mononucleotide adenyltransferase.

Results: BODIPY-deamido-NAD was isocratically eluted within 6 minutes using reverse-phase chromatography and its chromatographic peak was resolved. The calibration range, precision, and analytical recovery of the QUIN assay are suitable for the analysis of biological fluids. Compared with published quantitation limits for QUIN measurement by HPLC, this method is at least 30-fold more sensitive and has a lower limit of detection of 5.0 nmol/L. The sensitivity was comparable to that previously reported for gas chromatography/mass spectrometry (GC/MS) and the quantitation results of QUIN from samples of cerebrospinal fluid correlated well with that of the GC/MS method.

Conclusions: We established a novel method to quantify QUIN in biological samples. Due to its high sensitivity and the fact that it does not rely on MS instrumentation, this method has the potential for widespread adoption in research laboratories.

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生物样品中喹啉酸定量新方法的建立。

背景：喹啉酸（QUIN）在脑脊液和血清中的积累可作为多种神经精神和炎症性疾病的生物标志物。在这项研究中，我们开发了一种高灵敏度的方法来测量QUIN。方法：通过重组喹啉酸磷酸核糖基转移酶将QUIN转化为烟酸单核苷酸，再通过重组烟酸单核苷酸腺苷转移酶生成荧光（BODIPY）标记的脱酰胺- nad，建立反相高效液相色谱（HPLC）荧光检测方法。结果：BODIPY-deamido-NAD用反相色谱法在6分钟内被等压洗脱，并分离出色谱峰。该方法的校准范围、精密度和分析回收率适用于生物流体的分析。与已有的HPLC定量限相比，该方法的灵敏度提高了至少30倍，检测下限为5.0 nmol/L。灵敏度与先前报道的气相色谱/质谱（GC/MS）相当，脑脊液样品中QUIN的定量结果与GC/MS方法具有良好的相关性。结论：建立了一种新的定量生物样品奎因的方法。由于其高灵敏度和不依赖于质谱仪器的事实，该方法具有在研究实验室广泛采用的潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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