National and Kapodistrian University of Athens, School of Health Sciences, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Panepistimiopolis, 15771 Athens, Greece.
National Hellenic Research Foundation, Institute of Biology, Medicinal Chemistry and Biotechnology, 48 Vassileos Constantinou, 11635 Athens, Greece.
J AOAC Int. 2020 Jan 1;103(1):83-88. doi: 10.5740/jaoacint.18-0408.
The dietary supplementation of livestock with antioxidants to improve the meat quality represents an active research area of high commercial impact. In order to investigate the optimal dosing, analytical methodologies need to be developed in various tissues to evaluate which concentration does remain in the tissue.
We aimed to develop and validate a sensitive and specific methodology for the simultaneous quantitative determination of hesperidin, naringin, hesperetin, and naringenin in chicken tissue samples employing ultra-performance LC-tandem MS.
Lipid extraction using cold chloroform was performed followed by protein precipitation by cold acetone. Chromatography was performed on a C18 column using a ternary gradient of water, acetonitrile, and isopropanol-acetonitrile-acetone (58+40+2, v/v) as the mobile phase. Detection was performed by electrospray ionization in negative ion mode with the selected reaction monitoring technique.
Calibration plots exhibited good linearity (r2 > 0.99) over the concentration range from 0.125 to 25 μg/g tissue for the four analytes, and the lower LOQ for the four analytes was 0.125 μg/g tissue. The repeatability as percent relative SD and precision as percent accuracy were <20 and >80%, respectively.
The developed methodology was applied for the quantitative determination of hesperidin, naringin, hesperetin, and naringenin in tissue samples after dietary supplementation with 1.5 g/kg hesperidin and 1.5 g/kg naringin in Ross 308 broiler chickens.
This is the first methodology to access naringin, naringenin, hesperidin, and hesperetin in chicken tissue. It involved simple sample preparation, and the mass spectrometry based detection ensures high specificity and sensitivity.
在畜牧业中补充抗氧化剂以改善肉质是一个具有高商业影响力的活跃研究领域。为了研究最佳剂量，需要在各种组织中开发分析方法，以评估哪种浓度会残留在组织中。
我们旨在开发和验证一种灵敏且特异的超高效液相串联质谱法，同时定量测定鸡肉组织样品中的橙皮苷、柚皮苷、橙皮素和柚皮素。
采用冷氯仿进行脂质提取，然后用冷丙酮进行蛋白质沉淀。采用水、乙腈和异丙醇-乙腈-丙酮（58+40+2，v/v）的三元梯度作为流动相，在 C18 柱上进行色谱分离。采用电喷雾电离在负离子模式下，采用选择反应监测技术进行检测。
校准曲线在四种分析物的浓度范围为 0.125 至 25μg/g 组织时表现出良好的线性（r2>0.99），四种分析物的最低定量限均为 0.125μg/g 组织。重复性的相对标准偏差和精密度的准确度均<20%和>80%。
该方法已应用于在 Ross 308 肉鸡中以 1.5 g/kg 橙皮苷和 1.5 g/kg 柚皮苷的剂量进行饮食补充后，对组织样品中橙皮苷、柚皮苷、橙皮素和柚皮素的定量测定。
这是首次在鸡肉组织中检测到橙皮苷、柚皮苷、橙皮素和柚皮素的方法。它涉及简单的样品制备，基于质谱的检测确保了高特异性和灵敏度。
