Quantification By Gas Chromatography of the Content of Amino Acids Present in Sausages Fortified with Quinoa Vegetable Protein

The objective of this research was to determine the quality of the protein present in sausages fortified with quinoa as a substitute for animal protein, through the identification and quantification of amino acids, using gas chromatography and precolumn derivatization. The amino acid composition found in the analyzed products was predominantly composed of: Threonine (THR) with a concentration of 1046.32µmol / L, aminobutyric acid (ABA) with a concentration of 9685.68 µmol / L and glutamic acid (GLU) with a concentration of 1178.71 µmol / L. These values were found in the treatment with the highest percentage of quinoa flour, establishing a directly proportional relationship between the concentrations of these amino acids and the percentage of quinoa. Gas chromatography was an adequate technique for determining the amino acid profile due to its speed and sensitivity. Keywords: amino acids, sausages, quinoa, derivatization, gas chromatography. RESUMEN La presente investigación tiene por objetivo determinar la calidad de la proteína presente en embutidos fortificados con quinua como sustituyente de la proteína animal, a través de la identificación y cuantificación de aminoácidos mediante la aplicación de cromatografía de gases y la derivatización precolumna. La composición de aminoácidos encontrada en los productos analizados destaca la presencia mayoritaria de: Treonina (THR) con una concentración de 1046,32 µmol/L, ácido aminobutírico (ABA) con una concentración de 9685,68 µmol/L y ácido glutámico (GLU) con una concentración de 1178,71 µmol/L, todos estos valores se presentaron en el tratamiento con mayor porcentaje de harina de quinua estableciéndose una relación directamente proporcional entre las concentraciones de estos aminoácidos y el porcentaje de adición de quinua en los tratamientos estudiados. Se puede concluir que la cromatografía de gases empleada resultó una técnica adecuada para la determinación del perfil aminoacídico por la rapidez y sensibilidad presentada sobre las muestras estudiadas. Palabras claves: aminoácidos, embutidos, quinua, derivatización, cromatografía de gases.

Agriophyllum Oligosaccharides Ameliorate Diabetic Insulin Resistance Through INS-R/IRS/Glut4-Mediated Insulin Pathway in db/db Mice and MIN6 Cells

We have previously reported that Agriophyllum oligosaccharides (AOS) significantly enhance glycemic control by increasing the activation of insulin receptor (INS-R), insulin receptor substrate-2 (IRS-2), phosphatidylinositol 3 kinase (PI3K), protein kinase B (AKT), peroxisome proliferator-activated receptor (PPAR)-γ, and glucose transporter 4 (Glut4) proteins in hepatic tissues. However, the effect of glucose control by AOS on the regulation of pancreatic tissues in db/db mice and MIN6 cells remains to be determined. An oral dose of AOS (380 or 750 mg/kg) was administered to type-2 diabetic db/db mice for 8 weeks to determine whether AOS regulates glucose by the INS-R/IRS/Glut4-mediated insulin pathway. Meanwhile, the effects of AOS on glucose uptake and its related signaling pathway in MIN6 cells were also investigated. The results showed that the random blood glucose (RBG) level in the AOS-treated group was lower than that in the control group. AOS reduced the levels of glycated hemoglobin (HbA1c) and free fatty acid (FFA) and significantly improved the pathological changes in the pancreatic tissues in db/db mice. Moreover, immunohistochemical analysis revealed that the expression of INS-R, IRS-1, IRS-2, and Glut4 was increased in the AOS-treated group than in the model group. Further, in vitro experiments using MIN6 cells showed that AOS regulated INS-R, IRS-1, IRS-2, and Glut4 protein and mRNA levels and attenuated insulin resistance and cell apoptosis. The results of both in vitro and in vivo experiments were comparable. Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometric analysis of AOS with precolumn derivatization with 3-amino-9-ethylcarbazole (AEC) tentatively identified five types of sugars: glucose, lactose, rutinose, glucuronic acid, and maltotriose. Our present study clearly showed that AOS is efficacious in preventing hyperglycemia, possibly by increasing insulin sensitivity and improving IR by regulating the INS-R/IRS/Glut4 insulin signal pathway. Therefore, AOS may be considered as a potential drug for diabetes treatment.

Oenological Processes and Product Qualities in the Elaboration of Sparkling Wines Determine the Biogenic Amine Content

The biogenic amine (BA) content in wines is dependent on the fermentation processes and other oenological practices, as well as on grape quality. These compounds can participate in different cellular functions in humans; however, the intake of high amounts can provoke some toxicological effects. For that reason, controlling the evolution of biogenic amines in wine production processes is of extreme importance. This work aims to assess the occurrence of biogenic amines in sparkling wines and related samples, including musts, base wines, stabilized wines, and three-month and seven-month aged sparkling wines obtained from Pinot Noir and Xarel lo grape varieties. The determination of BA content relies on liquid chromatography with fluorescence detection (HPLC–FLD) with precolumn derivatization of analytes with dansyl chloride. The analysis has shown that putrescine is the most abundant amine in these types of samples. Ethanolamine, tyramine, spermine, and histamine concentrations are also remarkable. Principal component analysis has been applied to try to extract featured information concerning overall patterns dealing with wine production steps and qualities. Interesting conclusions have been drawn on BA formation depending on different factors. BA concentrations are quite low in must but rise, especially after the first alcoholic fermentation. Moreover, BA levels are much lower in the range of products elaborated with grapes of the best qualities while they significantly increase when using grapes of lower qualities. The results obtained pointed out the analytical potential of using BAs to control the quality of wine and its production processes, thus providing valuable information for both wineries and consumers.

Investigation of the α-Dicarbonyl Compounds in Some Snack Foods by HPLC Using Precolumn Derivatization with 4-Nitro-1,2-Phenylenediamine

Snack foods are widely consumed in today's modern diet. Food processing techniques and food composition may increase advanced glycation products (AGEs) in snack foods. The present study aimed to determine the most potent precursors of AGEs in snack foods. For this purpose, commonly consumed some snacks foods were obtained from markets in Istanbul, Turkey. The amount of α dicarbonyl compounds (α-DCs,) glyoxal (GO), and methylglyoxal (MGO) were determined by high-performance liquid chromatography. The measured amount of GO and MGO ranged between 4-684 µg / 100 g and 28-1573 µg / 100 g in snack foods, respectively. In our study, high levels of MGO were detected in wafer hazelnut chips with cheese and peanuts. Due to their high-fat content, the formation of GO and MGO may occur through lipid peroxidation. In addition, the fragmented state of hazelnuts and peanuts in samples may increase lipid peroxidation. Free sugar content in Turkish delight and cake with fruit might contribute to the α-DCs formation by caramelization reaction due to high temperature. In conclusion, snack products that are frequently consumed have many unfavorable features for health. It is important to limit snack food consumption in terms of reducing AGEs exposure.

Liquid Chromatographic Fingerprints for the Characterization of Flavanol-Rich Nutraceuticals Based on 4-Dimethylaminocinnamaldehyde Precolumn Derivatization

Flavanols consist of a great family of bioactive molecules displaying a wide range of health-promoting attributes for humans, including antioxidant, antimicrobial or anti-inflammatory effects. As a result, botanical species rich in this type of compound are often used to develop nutraceutical products or dietary supplements with recognized healthy attributes. This paper aims at characterizing nutraceutical products using liquid chromatographic fingerprints related to flavanol composition. Catechins and their oligomers were exploited to characterize and authenticate various commercial products prepared with extracts of red berries and medicinal plants. These compounds resulted in interesting descriptors of some fruits and vegetables, thus providing an additional perspective for the study of nutraceuticals. For such a purpose, a new method based on liquid chromatography with UV/Vis detection (HPLC–UV/Vis) with precolumn derivatization with 4-dimethylaminocinnamaldehyde was developed. Results indicated that the separation of flavanols was very complex due to the degradation of procyanidin derivatives. The resulting data sets were analyzed using chemometric methods such as principal component analysis and partial least square–discriminant analysis. Despite the complexity of chromatographic fingerprints, nutraceutical samples could be discriminated according to their main ingredients. In general, catechin and epicatechin were the most abundant compounds in the different samples, and procyanidin A2 was highly specific to cranberry.

Determination of residual dimethylaminopropylamine in aminopropylamides of fatty acid using high-performance liquid chromatography

A procedure for determination of the residual content of N,N-dimethyl-1,3-propanediamine (DMAPA) during synthesis of N,N-dimethylaminopropylamides of fatty acids (DMAPA FA) from coconut oil has been developed. The analysis was performed using high performance liquid chromatography on a Shimadzu LC-20 Prominence device equipped with a diode array detector and reversed phase column GL Sciences Inertsil ODS-3. Precolumn derivatization was carried out with a 13% — dansyl chloride (5-(dimethylamino)naphthalene-1-sulfonyl chloride) solution in acetone to increase the analyte response. The selected composition of the mobile phase — acetonitrile and triethylamine phosphate buffer solution (pH 3.0) in a volume ratio of 2:3 and optimized chromatographic conditions provided a clear peak of DMAPA with a retention time of 8.7 – 8.9 min. The proposed method provides determination of 0.02 – 10% DMAPA in DMAPAFA samples. Correctness of the procedure was confirmed in spike tests. The results obtained can be used for assessing the degree of conversion of starting materials in the synthesis of N,N-dimethylaminopropylamides of fatty acids, as well as for forecasting the content of N,N-dimethyl-1,3-propanediamine in the products on their base.

Development and Verification of a Precolumn Derivatization LC-MS/MS Method for the Pharmacokinetic Study of Houttuynine of Houttuynia Essential Oil

Houttuynia essential oil (HEO) has excellent antiviral, anti-inflammatory, and other pharmacological effects, but the lack of effective analytical methods to quantify HEO in plasma has hindered its better clinical monitoring. Houttuynine (Hou) is one of the main active ingredients and quality control substances of HEO, so the pharmacokinetic study of HEO could be conducted by determining Hou blood concentration. Hou is active and not stable in plasma, which makes its blood concentration difficult to measure. In this work, a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) method for Hou determination in rat blood was established that involves Hou being derivatized with 2, 4-dinitrophenylhydrazine to form a stable compound to prevent degradation. Herein, p-Tolualdehyde-2,4-dinitrophenylphenylhydrazone was selected as an internal standard substance and the LC-MS/MS method was evaluated for selectivity, precision, accuracy, calibration limit, matrix effect, recovery, and stability. Good linearity (r2 = 0.998) was reached in the range of 2–2000 ng/mL, and the lower limit of quantification of Hou was determined to be 2 ng/mL. The mean intra-assay accuracy ranged from 77.7% to 115.6%, whereas the intra-assay precision (relative standard deviation, RSD) was below 11.42%. The matrix effect value for Hou in rat plasma was greater than 75%, and for the internal standard (IS) it was 104.56% ± 3.62%. The extraction recovery of Hou were no less than 90%, and for the IS it was 96.50% ± 4.68%. Our method is sensitive and reliable and has been successfully applied to the pharmacokinetic analysis of Hou in rats given HEO via gavage and injection.

An Identification and Study of Amino Acids of Erigeron annuus Herb

Amino acids in the extract of Erigeron annuus herb were determined using an automatic precolumn derivatization with fluorenylmethyl-chloroformate and reversed-phase liquid chromatography with fluorescence and UV Vis detection. This objective is reached with automatic derivatization using o-phthalaldehyde (OPA) for primary amino acids and 9-Fluorenylmethyl chloroformate (FMOC) for secondary amino acids. Then derivatization integrates into high performance liquid chromatography (HPLC). The applied procedure is fast with easily reproduced results. The insufficient knowledge about amino acids composition of herb of Erigeron annuus is the basis for study. This work reports on content of 16 free and bound amino acids (391.41 μg/mg) in the plant raw material and influence’s evaluation of different extraction types on the amino acid profile. The total content of free amino acids was 4.66 μg/mg; proline prevailed (2.498 μg/mg). The total content of bound amino acids was 386.7 μg/mg; proline (146.8 μg/mg), arginine (67.8 μg/mg), phenylalanine (25.8 μg/mg), asparagine (24.3 μg/mg), histidine (20.4 μg/mg), alanine (18.2 μg/mg), serine (16.6 μg/mg), valine (16.0 μg/mg) were the dominant amino acids. Nine amino acids were classified as essential.