Quantification Of Trigonelline Research Articles

A Validated Trigonelline-Based Method for the Standardization and Quality Control of Trigonella foenum-graecum L.

Background Fenugreek, or Trigonella foenum-graecum L, is an edible and medicinal plant of the Fabaceae family. Fenugreek seeds contain a variety of phytochemicals, including proteins, lipids, amino acids, vitamins, flavonoids, steroidal saponins, coumarin, and alkaloids. Trigonelline TG is a bioactive plant alkaloid initially extracted from fenugreek seeds. A substantial portion of fenugreek’s health benefits may rely on the presence of TG. This study addresses the research gap for a fast, green, and economical method for quantifying trigonelline (TG) in fenugreek. Methods Fenugreek seeds from various origins were extracted using three green solvents: acetone (ACt), ethanol (EtOH), and water (H2O). The UPLCMSMS method was developed and validated using a green mobile phase of H2O: EtOH, and an r2-value of 0.999 in the linearity range of 0.1-500 ppb was adopted. The method was validated with an accuracy of 98.6% for trace analysis of TG using a small amount (10 mg) of fenugreek samples from five different origins. Results The average extract yield (±SD) was 5.36±6.3, with the highest extract yield observed in H2O. The ESI (+ve) of the UPLCMSMS resulted in the fragmentation pattern (m/z) 138→94.10→92.05→78.20. The TG quantification revealed an average TG concentration of 181.4, with the highest amount of TG in H2O extract (392.7±132.4 ppb), followed by EtOH (91.9±83.3 ppb) and ACt (59.5±30.9 ppb). The TG amount observed in the validation step substantiated the efficiency and reproducibility of the developed method. Conclusions The method may be used as an effective tool for a green, rapid, economical, and eco-friendly extraction and quantification of TG in diverse matrices of pharmaceutical, cosmeceutical, herbal, and food products.

Quality-controlled LC-ESI-MS food metabolomics of fenugreek (Trigonella foenum-graecum) sprouts: Insights into changes in primary and specialized metabolites

Fenugreek (Trigonella foenum-graecum L.) is an important food and spice with bioactive compounds against diabetes. In this study, fenugreek seeds germinating in darkness for 72 h were studied using quantification of trigonelline and 4-hydroxyisoleucine and an LC-ESI-MS/MS-based metabolomic approach capable of accurately estimating 237 features from various primary and specialized compound classes.During germination, the concentrations of trigonelline and 4-hydroxyisoleucine rose by 33.5% and 33.3%, respectively. At the same time, untargeted metabolomics revealed 9 putative flavonoids increasing 1.19- to 2.77-fold compared to the dormant seeds. A set of 19 steroid saponins rose by 1.08- to 31.86-fold. Primary metabolites however showed much more variability: abundance changes in amino acid derivatives, peptides and saccharides fell in the 0.09- to 22.25-fold, 0.93- to 478.79-fold and 0.36- to 941.58-fold ranges, respectively.To increase biosynthesis of specialized metabolites during germination, sprouts were exposed to 1–100 mM methyl jasmonate (MeJA) and methyl salicylate (MeSA). The hormone treatments affected normal metabolism: 67.1–83.1 % and 64.1–83.5 % of compounds showed a reduction compared to the controls in 100 mM MeJA and MeSA treatments at different sampling time points. Contrary to expectations, the abundance of flavonoids decreased, compared to the control sprouts (0.75- and 0.68-fold change medians, respectively). The same was observed for most, but not all steroid saponins.The quality-controlled untargeted metabolomics approach proved to yield excellent insight into the metabolic changes during germination of fenugreek. The results suggest that although fenugreek germination causes major shifts in plant metabolism, there are no major qualitative changes in bioactive specialized metabolites during the first three days. This stability likely translates into good bioactivity that is similar to that of the seeds. Because the large changes in the primary metabolites likely alter the nutritive value of the seed, further studies are warranted.

Standardization of Trigonella foenum-graecum L. Seeds: A Quality by Design Approach

Abstract: Aim: To standardize Trigonella foenum-graecum L. Seeds by developing QbD based HPLC method for identification and quantification of trigonelline in T. foenum graecum L. seeds, along with evaluation of various quality control parameters. Methods: The Analytical Target Profile and Critical Quality Attributes were determined followed by optimization of HPLC method by using 22 factorial design for designing the experiments for selected independent factors. Method Operable Design Region was developed for finding out the optimized chromatographic conditions. Further quality control parameters such as macroscopic and microscopic characters, physicochemical and phytochemical characterization including determination of toxic elements were carried out on the herb. Results: By application of QbD approach the optimized mobile phase was identified as water with 0.01% Hydrochloric acid and Methanol in the ratio of 70:30, with the flow rate of 1 mL/ min and UV detection at 263 nm. The linear model was established in the range of 2-10μg/mL with R2 value 0.998. The retention time of Trigonelline was found to be 2.877 min and the amount of Trigonelline in T. foenum-graecum L. Seeds was found to be 0.58%. The inter-day and intra-day precision were less than 2%, with accuracies between 96.6-110% of the true values. The quality control parameters showed the results within specified limits and the seeds showed absence of toxic elements in it. Conclusion: From the above finding we can conclude that the application of QbD approach for standardization of herbal drug can serve as an important tool for development of herbal drugs with desired quality. Key words: Quality by Design, Trigonella foenum-graecum L., Trigonelline, Standardization, HPLC.

Quantitative determination of trigonelline in mouse serum by means of hydrophilic interaction liquid chromatography-MS/MS analysis: Application to a pharmacokinetic study.

Trigonelline is a pyridine alkaloid found in fenugreek seeds and coffee beans. Most of the previous studies are concerned with the quantification of trigonelline along with other constituents in coffee herbs or beverages. Only a few have focused on its determination in animal or human tissues by applying different modes of HPLC with UV or MS detection. The aim of the study was to develop and validate a fast and simple method for trigonelline determination in serum by the use of hydrophilic interaction liquid chromatography (HILIC) with ESI-MS/MS detection. Separation of trigonelline was achieved on a Kinetex HILIC column operated at 35°C with acetonitrile-ammonium formate (10 mm, pH = 3) buffer mixture (55:45, v/v) as the mobile phase. The developed method was successfully applied to determine trigonelline concentration in mouse serum after intravenous administration of 10 mg/kg. The developed assay is sensitive (limit of detection = 1.5 ng/mL, limit of quantification = 5.0 ng/mL) and linear in a concentration range from 5.0 to 250.0 ng/mL. Sample preparation is limited to deproteinization, centrifugation and filtration. The application of the HILIC mode of chromatography with MS detection and selection of deuterated trigonelline as internal standard allowed a rapid and precise method of trigonelline quantification to be to developed.

Development of Microwave-Assisted Extraction of Trigonelline Biomarker fromTrigonella foenum-graecumSeeds Followed by High-Performance Thin-Layer Chromatographic and High-Performance Liquid Chromatographic Analyses

An efficient and fast microwave-assisted extraction (MAE) technique was developed for extracting trigonelline as an indicative biomarker for the quality control of Trigonella foenum-graecum seeds. The MAE procedure was optimized and compared with other conventional extraction techniques. The optimal conditions of MAE were 50% methanol as solvent, solid—liquid ratio 1:20 g mL−1, irradiation power 40%, and two extraction cycles, 3 min each. The proposed extraction technique produced a maximum yield of 7.89 mg g−1 trigonelline in 6 min which was 1.28 and 2.20 times more efficient than 3 h of heat reflux and 15 h of maceration extraction, respectively. Furthermore, rapid high-performance thin-layer chromatographic and high-performance liquid chromatographic methods for the quantification of trigonelline were established and validated. The methods were found to be simple, sensitive, precise, accurate, and specific for the estimation of trigonelline in T. foenum-graecum seeds extract and overcame disadvantages ...