Abstract
A new HPLC method was optimized and validated for the determination of asparagine in Asparagus lycicus plant, an endemic specie found in Turkey, and the phytochemical analysis of this plant was carried out. For asparagine analysis, optimum separation was obtained with water:methanol mixture (95:5, v/v) in the presence of formic acid (FA):ammonium formate (AF) buffer as mobile phase by using a C18 reversed phase column. In this new method, retention time (0.61 min) of asparagine was very short. The linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and selectivity parameters were studied for the validation of optimized method. By this method, satisfactory recovery (100.10 ± 2.86%) was obtained for asparagine analysis and LOD and LOQ values were determined to be 0.28 mg L−1 and 0.92 mg L−1, respectively. Freeze-thaw stability of the method is quite high. The results of matrix effect experiments showed that the matrix of Asparagus lycicus plant does not significantly affect the quantification analysis of asparagine. In the phytochemical characterization of endemic Asparagus lycicus plant, asparagine content (6.14 mg g-1 plant), total phenolic (13.22 mg GAE g-1 plant) and flavonoid (3.24 mg CE g-1 plant) contents, phenolic and flavonoid components (28.8 mg rosmarinic acid, 109.2 mg ferulic acid, 531.2 mg chlorogenic acid, 2.8 mg o-coumaric acid, 17.6 mg p-coumaric acid and 6.0 mg sinapic acid per kg plant), total antioxidant activity (13.2 mg mg-1 DPPH), vitamin C content (234.6 mg kg-1 plant), organic acid content as malic acid (2.9 mg kg-1 plant), sugar components (25.15 g fructose and 18.25 g glucose per kg plant), total anthocyanin content (5.15 mg kg-1 plant), total oil content (9.65%) and fatty acid components (oleic acid (55.36%), and linoleic acid (32.01%) and palmitic acid (7.61%)) were determined. The results of this study exhibited that endemic Asparagus lycicus plant might be considered as a promising natural source with antioxidant and bioactive constituents in the potential applications of pharmaceutical and food industries.
Similar content being viewed by others
References
Armstrong M, Jonscher K, Reisdorph NA (2007) Analysis of 25 underivatized amino acids in human plasma using ion-pairing reversed-phase liquid chromatography/time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 21:2717–2726
Arraiza MP, Gonzalez-Coloma A, Burillo J, Calderon-Guerrero C (2007) Medicinal and Aromatic Plants: The Basics of Industrial Application, first edn. Bentham Science Publishers, Sharjah
AOAC (2005) Official Methods of Analysis of AOAC, 16th edn. Association of Official Analytical Chemistry, Washington
Bai C, Reilly CC, Wood BW (2007) Identification and quantitation of asparagine and citrulline using high-performance liquid chromatography (HPLC). Anal Chem Insights 28:31–36
Bara L, Pop C, Horj E, Iordache A, Laslo C, Culea M (2010) Amino acids determination in meat by GC-MS. Bull UASVM Animal Sci Biotechnol 67:75–79
Bartolomeo MP, Maisano F (2006) Validation of a reversed-phase HPLC method for quantitative amino acid analysis. J Biomol Tech 17:131–137
Chen XH, Ma LH, Dong YW, Song H, Pu Y, Zhou QY (2017) Evaluation of the differences in phenolic compounds and antioxidant activities of five green asparagus (Asparagus officinalis L.) cultivars. Qual Assur Saf Crop 9:1–10
Christaki E, Bonos E, Giannenas I, Florou-Paneri P (2012) Aromatic plants as a source of bioactive compounds. Agriculture. 2:228–243
Cruz AF, Barbosa TMCC, Adelino TER, Lima WP, Mendes MO, Valadares ER (2016) Amino acid reference intervals by high performance liquid chromatography in plasma sample of Brazilian children. J Bras Patol Med Lab 52:70–77
Danowska-Oziewicz M, Narwojsz A, Draszanowska A, Marat N (2020) The effects of cooking method on selected quality traits of broccoli and green asparagus. Int J Food Sci Technol 55:127–135
Esetlili BÇ, Pekcan T, Çobanoğlu Ö, Aydoğdu E, Turan S, Anaç D (2014) Essential plant nutrients and heavy metals concentrations of some medicinal and aromatic plants. J Agric Sci 20:239–247
Fan R, Yuan F, Wang N, Gao Y (2015) Extraction and analysis of antioxidant compounds from the residues of Asparagus officinalis L. J Food Sci Technol 52:2690–2700
Farkas T, Toulouee J (2003) Asparagine analysis in food products. LG GC Eur 21:14–16
Fischer UA, Carle R, Kammerer DR (2011) Identification and quantification of phenolic compounds from pomegranate (Punica granatum L.) peel, mesocarp, aril and differently produced juices by HPLC-DAD-ESI/MS. Food Chem 127:807–821
Güneş F (2018) An ethnobotany study in Enez town from Edirne. CUPMAP. 1:28–35
Han Y, Du J, Li J, Li M (2019) Quantification of the organic acids in hawthorn wine: A comparison of two HPLC methods. Molecules 24:2150–2164
Jeong HC, Kim T, Yang DH, Shin KH (2018) Development of a UPLC-MS/MS method for the therapeutic monitoring of L-asparaginase. Transl Clin Pharmacol 26:134–140
Keith LH, Crummet W, Deegan J, Libby RA, Taylor JK, Wentler G (1983) Principles of environmental analysis. Anal Chem 55:2210–2218
Kennedy A, Bivens A (2017). Methods for the analysis of underivatized amino acids by LC-MS. Agilent Technologies Application Note. 1-4.
Kim BY, Cui ZG, Lee SR, Kim SJ, Kang HK, Lee YK, Park DB (2009) Effects of Asparagus officinalis extracts on liver cell toxicity and ethanol metabolism. J Food Sci 74:204–208
Lafka TI, Sinanoglou V, Lazos ES (2007) On the extraction and antioxidant activity of phenolic compounds from winery wastes. Food Chem 104:1206–1214
Le A, Ng A, Kwan T, Cusmano-Ozog K, Cowan TM (2014) A rapid, sensitive method for quantitative analysis of underivatized amino acids by liquid chromatography-tandem mass spectrometry. J Chromatogr B 944:166–174
Nemkov T, D’Alessandro A, Hansen KC (2015) Three-minute method for amino acid analysis by UHPLC and high-resolution quadrupole orbitrap mass spectrometry. Amino Acids 47:2345–2357
Nguyen TVL, Tran TYN, Lam DT, Bach LG, Nguyen DC (2019) Effects of microwave blanching conditions on the quality of green asparagus (Asparagus officinalis L.) butt segment. Food Sci Nutr 7:3513–3519
Nielsen NJ, Granby K, Hedegaard RV, Skibsted LH (2006) A liquid chromatography – tandem mass spectrometry method for simultaneous analysis of acrylamide and the precursors, asparagine and reducing sugars in bread. Anal Chim Acta 557:211–220
Palfi M, Jurkovic Z, Cosic J, Tomic-Obrdalj H, Jurkovic V, Knezevic N, Vrandecic K (2017) Total polyphenol content and antioxidant activity of wild and cultivated asparagus in Croatia. Poljoprivreda/Agriculture. 23:56–62
Parpinello GP, Versari A (2000) A simple high-performance liquid chromatography method for the analysis of glucose, glycerol, and methanol in a bioprocess. J Chromatogr Sci 38:259–261
Pekal A, Pyrzynska K (2014) Evaluation of aluminium complexation reaction for flavonoid content assay. Food Anal Methods 7:1776–1782
Peters FT, Drummer OH, Musshoff F (2007) Validation of new methods. Forensic Sci Int 165:216–224
SANCO/12495/2011 (2011) Method validation and quality control procedures for pesticide residues analysis in food and feed. National Food Administration, Uppsala, Sweden
Shabir GA (2004) A practical approach to validation of HPLC methods under current good manufacturing practices. J Valid Technol 2:29–37
Shin EC, Hwang CE, Lee BW, Kim HT, Ko JM, Baek IY, Lee YB, Choi JS, Cho EJ, Seo WT, Cho KM (2012) Chemometric approach to fatty acid profiles in soybean cultivars by principal component analysis (PCA). Prev Nutr Food Sci 17:184–191
Slatnar A, Mikulic-Petkovsek M, Stampar F, Veberic R, Horvat J, Jakse M, Sircelj H (2018) Game of tones: sugars, organic acids, and phenolics in green and purple asparagus (Asparagus officinalis L.) cultivars. Turk J Agric For 42:55–66
Spanos GA, Wrolstad RE (1990) Influence of processing and storage on the phenolic composition of Thompson seedless grape juice. J Agric Food Chem 38:1565–1571
Sreedevi V, Kumar PR, Thatavarti R (2011) LC-MS method development and validation for the estimation of Felodipine in human plasma and stability studies of freeze thaw analyte. IJPSR. 2:65–73
Symes A, Shavandi A, Zhang H, Ahmed IAM, Al-Juhaimi FY, Bekhit AEDA (2018) Antioxidant activities and caffeic acid content in New Zealand Asparagus (Asparagus officinalis) roots extracts. Antioxidants. 7:1–13
Tojo M, Murakami M, Nagata Y (2012) Simple and low-cost high-performance liquid chromatographic method for determination of D- and L-amino acids. J Chromatogr Sci 50:393–395
Ullah F, Kang M, Khattak MK, Wahab S (2018) Experimentally investigated the asparagus (Asparagus officinalis L.) drying with flat-plate collector under the natural convection indirect solar dryer. Food Sci Nutr 6:1357
Veena N, Arora S, Kapila S, Singh RRB, Katara A, Pandey MM, Rastogi S, Rawat AKS (2014) Immunomodulatory and antioxidative potential of milk fortified with Asparagus racemosus. J Med Plants Stud 2:13–19
Whitmire M, Ammerman J, Lisio PD (2011) Bioanalysis method development, validation and sample analysis: Points to consider when conducting nonclinical and clinical studies in accordance with current regulatory guidances. J Anal Bioanal Technol 1:1–10
Zapata S, Dufour J (1992) Ascorbic, dehydroascorbic and isoascorbic acid simultaneous determination by reverse phase ion interaction HPLC. J Food Sci 57:506–511
Zhang H, Birch J, Xie C, Yang H, Dias G, Kong L, Bekhit AED (2018) Optimization of extraction parameters of antioxidant activity of extracts from New Zealand and Chinese Asparagus officinalis L root cultivars. Ind Crop Prod 119:191–200
Zhang H, Birch J, Xie C, Yang H, Bekhit AED (2019a) Optimization of ultrasound assisted extraction method for phytochemical compounds and in-vitro antioxidant activity of New Zealand and China Asparagus cultivars (officinalis L.) roots extracts. Food Chem 294:276–284
Zhang H, Birch J, Pei J, Ahmed IAM, Yang H, Dias G, El-Aty AMA, Bekhit AED (2019b) Identification of six phytochemical compounds from Asparagus officinalis L. root cultivars from New Zealand and China using UAE-SPE-UPLC-MS/MS: effects of extracts on H2O2-induced oxidative stress. Nutrients. 11:1–17
Acknowledgements
This work was supported by the Scientific Research Projects Coordination Unit of Akdeniz University (Project Number: FYL-2018-3119). We would like to thank the Bati Akdeniz Agricultural Research Institute (Antalya, Turkey) for HPLC, UV-Vis spectrophotometer, and GC-MS analyses.
Funding
This work was funded by the Scientific Research Projects Coordination Unit of Akdeniz University (Project Number: FYL-2018-3119).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics Approval
This article does not contain any studies using human or animals.
Informed Consent
Informed consent is not applicable.
Conflict of Interest
Orçun Çınar declares that he has no conflict of interest. Osman Duman declares that he has no conflict of interest. Sibel Tunç declares that she has no conflict of interest.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Çınar, O., Duman, O. & Tunç, S. Optimization and Validation of a New HPLC Method for the Determination of Asparagine Active Ingredient in Asparagus lycicus and Phytochemical Characterization of Endemic Asparagus lycicus Specie. Food Anal. Methods 14, 2003–2016 (2021). https://doi.org/10.1007/s12161-021-02017-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-021-02017-0