Development and validation of a method for quantitative determination of the amount of fructosans in chicory roots

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Introduction. To develop regulatory documentation for raw materials of chicory roots, it is necessary to standardize the raw materials according to the content of the main active substance, which is fructosans (the main substance is inulin) [1, 2]. The Russian Pharmacopoeia 15th edition, includes articles ФС.2.5.0070.18, ФС.2.5.0025.15, which include a method for determining the amount of fructosans in a plant, based on the spectrophotometric determination of the absorption of a colored complex of acid hydrolysis products of fructosans - 5-hydroxymethylfurfural (5-HMF) with resorcinol. However, in the scientific literature there are methods based on determining the absorption of hydrolysis products without additional complex formation.

The aim – development and validation of a method for the quantitative determination of the amount of fructosans in terms of inulin in chicory roots with spectrophotometric determination of 5-HMF absorption, without the use of additional reagents (resorcinol, thiourea).

Material and methods. The object of the study is the roots of common chicory. The development and validation of the method for the quantitative determination of the sum of fructosans in terms of inulin was carried out by spectrophotometry.

Results. The developed method has been positively evaluated in the following parameters: specificity (coincidence of absorption maxima at a wavelength of (286±2) nm); linearity (correlation coefficient 0.9987); accuracy (range of recovery percentage – from 96.59% to 103.50% and has an average value of 99.97%); repeatability (coefficient of variation (RSD) 1.52% < 2.00%); reproducibility (coefficients of variation of intralaboratory reproducibility do not exceed 2%).

Conclusion. The obtained results allow us to consider the developed method suitable for reliable quantitative determination of the amount of fructosans in terms of inulin in the roots of common chicory.

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作者简介

А. Gulenkov

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

编辑信件的主要联系方式.
Email: gulenkov@vilarnii.ru
ORCID iD: 0000-0002-1532-9330
SPIN 代码: 9436-3210

Ph.D. (Pharm), Leading Researcher, Department of Chemistry of Natural Compounds

俄罗斯联邦, 7 Grin str., Moscow, 117216

O. Kulyak

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants; Lomonosov Moscow State University

Email: gulenkov@vilarnii.ru
ORCID iD: 0000-0003-0385-3474
SPIN 代码: 9697-1220

Ph.D. (Pharm), Leading Researcher, Department of Chemistry of Natural Compounds, Head of the Testing Center, Associate Professor of the Department of Pharmaceutical Chemistry and Organization of Pharmaceutical Affairs of the Medical Scientific and Educational Institute

俄罗斯联邦, 7 Grin str., Moscow, 117216; Leninskie Gory 1, Moscow, 119991

V. Dul

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: gulenkov@vilarnii.ru
ORCID iD: 0000-0001-7824-4417
SPIN 代码: 8192-5803

Ph.D. (Pharm.), Leading Researcher, Department of Chemistry of Natural Compounds

俄罗斯联邦, 7 Grin str., Moscow, 117216

P. Mavrina

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: gulenkov@vilarnii.ru
ORCID iD: 0000-0002-8559-8771
SPIN 代码: 7119-8667

Post-graduate Student, Junior Researcher, Department of Chemistry of Natural Compounds

俄罗斯联邦, 7 Grin str., Moscow, 117216

I. Zotov

D. Mendeleev University of Chemical Technology of Russia

Email: gulenkov@vilarnii.ru
ORCID iD: 0009-0005-0705-8672

Student of the Faculty of Chemical and Pharmaceutical Technologies and Biomedical Preparations, Department of Chemistry and Technology of Biomedical Drugs

俄罗斯联邦, Miusskaya square 9, Moscow, 125047

A. Tomilina

D. Mendeleev University of Chemical Technology of Russia

Email: gulenkov@vilarnii.ru
ORCID iD: 0009-0009-7751-4310

Student of the Faculty of Chemical and Pharmaceutical Technologies and Biomedical Preparations, Department of Chemistry and Technology of Biomedical Drugs

俄罗斯联邦, Miusskaya square 9, Moscow, 125047

P. Mizina

All-Russian Scientific Research Institute of Medicinal and Aromatic Plants

Email: gulenkov@vilarnii.ru
ORCID iD: 0000-0001-6510-9603
SPIN 代码: 7084-1962

Dr.Sc. (Pharm.), Professor, Advisor to the Director, Honored Scientist of the Russian Federation

俄罗斯联邦, 7 Grin str., Moscow, 117216

参考

  1. Vyutnova O.M., Novikova I.A. Chemical composition of root chicory. Vegetable crops of Russia. 2019; (1): 83–85. (In Russian).
  2. Wan X., Guo H., Liang Y. et al. The physiological functions and pharmaceutical applications of inulin: A review. Carbohydrate Polymers. 2020. 246: 116589. doi: 10.1016/j.carbpol.2020.116589.
  3. Oleynikov D.N., Tankhaeva L.M. Study of the colorimetric reaction of fructose with resorcinol depending on the conditions of its implementation. Khimiya Rastitel’nogo Syr’ya [Chemistry of plant raw material]. 2008; 2: 69–74. (In Russian).
  4. Kopytko Ya.F. Determination of total carbohydrates (recalculated for fructose upon conversion to furans) in burdock juice. Khimiko-Farmatsevticheskii Zhurnal. 2017; 51(4): 38–40. (In Russian).
  5. Bokov D.O. Hromchenkova E.P., Sokurenko M.S. et.al. Development of a technique for the determination of inulin in natural instant chicory after enzymatic hydrolysis by high-performance liquid chromatography. Voprosy pitaniia [Problems of Nutrition]. 2017; 86(5): 50–55. (In Russian).
  6. Dyakova N.A. Development and validation of the express technique of allocation and quantitative definition of water-soluble polysaccharides of leaves of Arctium Lappa L. Khimiya Rastitel’nogo Syr’ya [Chemistry of plant raw material]. 2018; 4: 81–87. (In Russian).

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2. Fig. 1. Complexation reaction of inulin hydrolysis products with resorcinol

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3. Fig. 2. Linear dependence of optical absorption of 5-HMF (a) and complex with resorcinol (б) on the concentration of inulin CO

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4. Fig. 3. Graphs of UV spectra of CO inulin and fructosans of chicory roots after acid hydrolysis

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5. Fig. 4. Results of the convergence analysis for inulin CO (n=6): А – on the day of reagent preparation (CO solution and hydrochloric acid solution); Б – 4 days after solution preparation; В – 9 days after solution preparation; Г – 9 days of inulin CO solution, freshly prepared hydrochloric acid solution; Д – freshly prepared inulin CO solution, 9 days of hydrochloric acid solution; Е – freshly prepared inulin CO solution and hydrochloric acid solution

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6. Fig. 5. Results of determination of the content of total fructosans in 46 samples of chicory roots using a validated method

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