Figueiredo Ingrid Lima, Claus Thiago, Oliveira Santos Júnior Oscar, Almeida Vitor Cinque, Magon Thiago, Visentainer Jesuí Vergilio
State University of Maringa, Department of Chemistry, Av. Colombo, 5790, 87020-900 Maringa, Paraná State, Brazil.
State University of Maringa, Department of Chemistry, Av. Colombo, 5790, 87020-900 Maringa, Paraná State, Brazil.
J Chromatogr A. 2016 Jul 22;1456:235-41. doi: 10.1016/j.chroma.2016.06.012. Epub 2016 Jun 4.
In order to analyze the composition of fatty acids employing gas chromatography as the separation method, a derivatization of lipids using esterification and transesterification reactions is needed. The methodologies currently available are time consuming and use large amounts of sample and reagents. Thus, this work proposes a new procedure to carry out the derivatization of fatty acids without the need for prior extraction of lipids. The use of small amounts of sample (100mg) allows the analysis to be performed in specific parts of animals, in most cases without having them slaughtered. Another benefit is the use of small amounts of reagents (only 2mL of NaOH/Methanol and H2SO4/Methanol). The use of an experimental design procedure (Design Expert software) allows the optimization of the alkaline and acid reaction times. The procedure was validated for five minutes in both steps. The method was validated for bovine fat, beef, chicken, pork, fish and shrimp meats. The results for the merit figures of accuracy (from 101.07% to 109.18%), precision (RSDintra-day (from 0.65 to 3.93%), RSDinter-day (from 1.57 to 5.22%)), linearity (R(2)=0.9864) and robustness confirmed that the new method is satisfactory within the linear range of 2-30% of lipids in the sample. Besides the benefits of minimizing the amount of samples and reagents, the procedure enables gas chromatography sample preparation in a very short time compared with traditional procedures.
为了采用气相色谱作为分离方法来分析脂肪酸的组成,需要通过酯化和酯交换反应对脂质进行衍生化处理。目前可用的方法耗时较长,且需要大量的样品和试剂。因此,本研究提出了一种新的方法来进行脂肪酸的衍生化,无需事先提取脂质。使用少量样品(100mg)可以在动物的特定部位进行分析,在大多数情况下无需宰杀动物。另一个优点是使用少量试剂(仅2mL氢氧化钠/甲醇和硫酸/甲醇)。使用实验设计程序(Design Expert软件)可以优化碱性和酸性反应时间。该程序在两个步骤中均经过5分钟的验证。该方法已在牛脂、牛肉、鸡肉、猪肉、鱼肉和虾肉中得到验证。准确度(101.07%至109.18%)、精密度(日内相对标准偏差(RSDintra-day,0.65至3.93%),日间相对标准偏差(RSDinter-day,1.57至5.22%))、线性(R(2)=0.9864)和稳健性等指标的结果证实,该新方法在样品中脂质含量为2-30%的线性范围内是令人满意的。除了将样品和试剂用量降至最低的优点外,与传统方法相比,该程序能够在极短的时间内完成气相色谱样品制备。