National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, MS 38677, USA.
Natural Products Utilization Research Unit, Agricultural Research Service, United States Department of Agriculture, University, MS 38677, USA.
Molecules. 2022 Nov 2;27(21):7441. doi: 10.3390/molecules27217441.
Sample preparation remains both a challenging and time-consuming process in the field of bioanalytical chemistry. Many traditional techniques often require multi-step processes, which can introduce additional errors to the analytical method. Given the complexity of many biological matrices, thorough analyte extraction presents a major challenge to researchers. In the present study, a headspace solid-phase microextraction (HS-SPME) coupled with a GC/Q-ToF-MS method, was developed to quantify in vitro metabolism of β-caryophyllene by both human liver microsome (HLM) and S9 liver fractions. Validation of the method was demonstrated both in terms of linearity ( = 0.9948) and sensitivity with a limit of detection of 3 ng/mL and a limit of quantitation of 10 ng/mL. In addition, the method also demonstrated both inter- and intra-day precision with the relative standard deviation (RSD) being less than 10% with four concentrations ranging from 50-500 ng/mL. Since this method requires no solvents and minimal sample preparation, it provides a rapid and economical alternative to traditional extraction techniques. The method also eliminates the need to remove salts or buffers, which are commonly present in biological matrices. Although this method was developed to quantify in vitro metabolism of one analyte, it could easily be adapted to detect or quantify numerous volatiles and/or semi-volatiles found in biological matrices.
样品制备在生物分析化学领域仍然是一个具有挑战性和耗时的过程。许多传统技术通常需要多步过程，这可能会给分析方法引入额外的误差。鉴于许多生物基质的复杂性，彻底提取分析物对研究人员来说是一个主要挑战。在本研究中，开发了一种顶空固相微萃取（HS-SPME）与 GC/Q-ToF-MS 方法，用于定量人肝微粒体（HLM）和 S9 肝部分体外代谢β-石竹烯。该方法的验证在线性（ = 0.9948）和灵敏度方面均得到了证明，检测限为 3ng/mL，定量限为 10ng/mL。此外，该方法还表现出日内和日间精密度，四个浓度范围为 50-500ng/mL 时相对标准偏差（RSD）小于 10%。由于该方法不需要溶剂且样品制备量最小，因此提供了一种比传统提取技术更快、更经济的替代方法。该方法还消除了去除盐或缓冲液的需要，这些物质通常存在于生物基质中。尽管该方法是为定量分析一种分析物的体外代谢而开发的，但它可以很容易地适应检测或定量生物基质中存在的许多挥发性和/或半挥发性物质。
