Free amino acids, biogenic amines, and ammonium profiling in tobacco from different geographical origins using microwave‐assisted extraction followed by ultra high performance liquid chromatography

Abstract

This work describes a rapid, stable, and accurate method for determining the free amino acids, biogenic amines, and ammonium in tobacco. The target analytes were extracted with microwave-assisted extraction and then derivatized with diethyl ethoxymethylenemalonate, followed by ultra high performance liquid chromatography analysis. The experimental design used to optimize the microwave-assisted extraction conditions showed that the optimal extraction time was 10 min with a temperature of 60°C. The stability of aminoenone derivatives was improved by keeping the pH near 9.0, and there was no obvious degradation during the 80°C heating and room temperature storage. Under optimal conditions, this method showed good linearity (R > 0.999) and sensitivity (limits of detection 0.010-0.081 μg/mL). The extraction recoveries were between 88.4 and 106.5%, while the repeatability and reproducibility ranged from 0.48 to 5.12% and from 1.56 to 6.52%, respectively. The newly developed method was employed to analyze the tobacco from different geographical origins. Principal component analysis showed that four geographical origins of tobacco could be clearly distinguished and that each had their characteristic components. The proposed method also showed great potential for further investigations on nitrogen metabolism in plants.

Comprehensive characterization and quantification of multiple components in Dan‐Huang‐Qu‐Yu capsule using a multivariate data processing approach based on microwave‐assisted extraction with UHPLC and Q Exactive quadrupole‐orbitrap high‐resolution mass spectrometry

“…Hence, a new technique characterized by efficient use of organic solvent, time, and energy should be employed in the sample preparation procedure. Fortunately, MAE was finally found to possess all the virtues shown above and had been employed in our research with a high efficiency [10,11]. Moreover, Chinese medicine is composed of various chemical constituents, and therefore it is still a problem to have a synthesis evaluation after the determination of many interesting bioactive ingredients.…”

Alteration of S‐adenosylhomocysteine levels affects lignin biosynthesis in switchgrass

“…For amino acid profiling analysis, 500 mg of powdered tissue was extracted and derivatized with diethyl ethoxymethylenemalonate as described by Cai et al . (). The aminoenone derivatives were identified and quantified by liquid chromatography coupled to tandem mass spectrometry (LC‐MS/MS).…”

Nitrogen application and differences in leaf number retained after topping affect the tobacco (Nicotiana tabacum) transcriptome and metabolome

“…We followed the tobacco industry standards of the People’s Republic of China for protein determination by the continuous-flow method (YC/T 249–2008) to extract protein for subsequent content determination and soluble protein content using a commercial assay kit (Coomassie Brilliant Blue G-250, Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s instructions. The 21 free amino acid contents were measured by using on thin layer chromatography plates (TLC) as described previously [ 50 ]. We measured PAs content (isoamylamine, spermidine, phenethylamine, and putrescine) in tobacco leaves as described previously [ 51 ].…”