Development of techniques for quantitative analysis of lime flowers
Keywords:
Lime Flowers (Tilia cordata Mill.), Quantitative Analysis, Essential Oil, Flavonoids, Difluorochloromethane.Abstract
Introduction. The article is devoted to the development of techniques for quantitative analysis of lime flower in order to make amendments to existing pharmacopoeian monographs for this herbal drug. Lime inflorescences contain lipophilic biologically active substances (BAS) causing notable antimicrobial and anti-inflammatory effects and also more polar phenolic compounds with antiulcer activity. Considering this, it’s necessary to regulate all these groups of BAS quantitatively. Materials and methods. For this study six batches of lime flowers harvested in 2008-2009 yrs. in Kharkiv, Rivno and Zhitomir regions were used as crude herbal drug. Loss on drying was determined by routine pharmacopoeian procedures. Total content of lipophilic substances was determined gravimetrically after Soxhlet extraction of samples 1, 5, 7 and 10 g in weight with methylene chloride, considering that by its extracting ability this solvent is close to liquefied difluorochloromethane (freon R22) used by us for obtaining of lipophilic complexes. The duration of complete analytical extraction was determined by infusion of six 10 g assays of lime flowers during 1, 2, 3, 4, 5, 6 hours, then quantity of lipophilic extractives was revealed gravimetrically. Quantity of essential oil in lime flowers was evaluated under the procedure of ЕР7, 2.8.12. Weight of the herbal drug sample was 200 g, distillation rate – 2,5- 3,5 ml/min, volume of distillation liquid (water) – 500 ml, volume of xylene in the graduated tube – 0,50 ml.Total flavonoid content recalculated to quercetin was determined after hydrolysis with acidified acetone, withdrawing of flavonoid aglycones with ethylacetate and by further spectrophotometry of their complexes with aluminium chloride. All quantitative determinations were replicated five times for each assay. All chemicals and reagents were of analytical grade. Results and discussion. It was found that adequate accuracy of the analysis of lipophilic extractives taken with methylene chloride was achieved if assay weight is not less than 7 g: relative error was less than 2 %. Study of extraction kinetics showed that after 4 hours lipophilic substances soluble in methylene chloride were taken almost completely. Analyzing 6 batches of lime flowers from different Ukrainian regions, it was found that yield of total extractives taken with methylene chloride did fall within insignificant limits – 1,86-2,26 %, that testifies onto rather stable accumulation of these biologically active substances in lime flowers even despite different climatic conditions. Essential oil content varied in rather wide limits – 0,34-0,55 ml/kg of herbal drug and considerably depended on harvest place: in the batches from Northwest of Ukraine this value was less than in the Eastern ones. Year of harvesting had less influence on quantity of essential oil in lime flowers. The relative error of the quantitative determinations was within 4–8 %.Total flavonoid content in the analyzed batches of lime flowers was considerably variable (from 0,30 to 0,52 %) and depended mainly on place of harvesting, and this crude herbal drug from the north-west of Ukraine (Rivno region) showed the highest results among the analyzed samples. Besides, if consider the lower limit regulated by the monograph of Belarusian Pharmacopoeia, p. 370, then only one batch conformed to the norm (not less than 0,5 %). Therefore it’s recommended to fix the lower limit 0,3 % for content of total flavonoids in the corresponding monograph draft or analytical normative documentation. The relative error of the quantitative determinations of flavonoids was within 1,5–3,1 %.Conclusion.Six batchesof lime flowers harvested in various regions of Ukraine in 2008 and 2009 yrs. were investigated. Based on the results obtained it is offered to regulate the following quantity parameters in the amended monograph draft for lime flower: total extractives taken with methylene chloride – not less than 1,8 %, essential oil content – not less than 0,3 ml/kg, total flavonoid content recalculated as quercetin - not less than 0,3 %.
References
Matters of introduction of monographs on herbal drugs in the State Pharmacopoeia of Ukrai- ne / A. I. Grisodub, G. V. Georgiyevsky, T. M. Tikhonenko, V. P. Georgiyevsky // Pharmacom. – 2004. – № 4. – P. 3–17.
To the matter of introduction in the State Pharmacopoeia of Ukraine of general monographs on the herbal drugs and herbal drug preparations / E. K. Tovmasyan, A. G. Kotov, A. I. Grisodub, V. P. Georgiyevsky // Pharmacom. – 2004. – № 4. – P. 17–23.
Matters of «Lime flower» monograph introduction to the State Pharmacopoeia of Ukraine // A. G. Kotov, E. E. Kotova, T. M. Tikhonenko, V. G. Volovic // Pharmacom. – 2005. – № 1. – P. 54–59.
Kotov, A. G. Study of development and introduction of monographs on herbal drugs into the State Pharmacopoeia of Ukraine / A. G. Kotov // Pharmacom. – 2009. – № 1. – P. 5–19.
Matters of the standardization of motherwort herb and drugs on its base / E. E. Kotova, N. I. Tikhonenko, A. G. Kotov et al. // Pharmacom. – 2006. – № 4. – P. 50–58.
European Pharmacopoeia. – 7th ed. – Strasbourg : Council of Europe, 2005. – 3310 p.
State Pharmacopoeia of Belarus : in 3 vol. Vol. 2. General and specific monographs / Committee for Pharmaceutical and Microbiological Industry. – Minsk, 2009. – 471 p.
State Pharmacopoeia of Ukraine / State Enterprise “Scientific and Expert Pharmacopoeial Center”. – 1st Ed., Suppl.4. – Kharkiv, 2011. – 540 p.
British Pharmacopoeia 2009. – Bournemouth: Crown Publishers, 2008. – 10952 p.
State Pharmacopoeia of the USSR / Ministry of Public Health. – 11th Ed. – Moscow : Medicina, 1989. – Vol. 2. General methods of analysis. Herbal drugs. – 400 p.
Japanese Pharmacopoeia. – 15th Ed. – Tokio, 2006. – 1788 p.
Study of standardization criteria of herbal drug of some species of Lamiaceae Lindl. family at the USSR pharmacopoeias for the development of monographs for the State Pharmacopoeia of Ukraine / N. I. Tykhonenko, A. G. Vovk, A. G. Kotov et al. // Management, economy and quality assurance in pharmacy. – 2010. – Vol. 13, № 5. – Р. 4–15.
Dashutina, S. L. To the matter of the standardization of Chelidonium majus L. herb / S. L. Dashutina, A. G. Kotov, V. P. Georgiyevsky // Pharmacom. – 2005. – № 2/3. – Р. 134–140.
Matters of introduction into SPU of the monograph “Yarrow” / E. E. Kotova, I. S. Lukyanova, A. G. Kotov et al. // Pharmacom. – 2007. – № 2. – Р. 33–40.
Gryzodub, A. I. Characteristics of pharmacopoeial approaches to quantifying of raw herbal drugs and combined herbal drugs / A. I. Gryzodub, O. A. Evtifeeva, K. I. Proskurina // Pharma-com. – 2012. – № 3. – Р. 7–30.
Demyanenko, D. V. Microbiological study of liquefied-gas and supercritical extracts from lime flowers / D. V. Demyanenko, Ye. M. Babich, N. I. Sklyar // Pharmaceutical review. – 2012. – № 1. – Р. 119–123.
Pharmacological study of Freon extract from lime flowers in adjuvant-induced arthritis model in rats / S. M. Drogovoz, D. V. Demyanenko, G. V. Belik // Phytotherapy review. – 2013. – № 3. – Р. 41–44.
Patent 93003 UA, IPC (2013.01) A61K 36 / 00, A61K 31 / 33. Method of complex processing of linden blossom / Demyanenko V. G., Demyanenko D. V. Breusova S. V. (UA); National University of Pharmacy (UA). – appl. 21.05.08; publ. 27.12.10. – 6 p.
State Pharmacopoeia of Ukraine / State Enterprise “Scientific and Expert Pharmacopoeial Center”. – 1st Ed., Suppl.2. –Kharkiv, 2008. – 620 p.
Flavonoid content assay: Prevalidation and application on Plantago L. species / R. Jurisic Grubesic, J. Vukovic, D. Kremer, S. Vladimir-Knezevic // Acta Chim. Slov. – 2007. – Vol. 54, Iss. 2. – P. 397–406.
Breusova, S. V. Development of composition and technology of syrup on basis of phenolic complex from lime flowers : PhD Thesis : 15.00.01 / Breusova Svitlana Victorivna. – Kharkiv : National University of Pharmacy, 2009. – 167 p.
Demyanenko, D. V. Use of liquefied gases for extraction of biologically active substances from Tilia flos / D. V. Demyanenko, D. I. Dmitriyevsky // Proc. of the VIIth National Congress of Pharmacists of Ukraine, Kharkiv, 15 – 17th of September 2010 yr. – In 2 volumes. – Kharkiv : National University of Pharmacy, 2010. – Vol. 1. – P. 248.
Demyanenko, D. V. Research of process dynamics during extraction of lipophilic compounds from Tilia cordata inflorescences / D. V. Demyanenko // Current Issues in Pharmacy and Medicine Science and Practice. – 2011. Vol. XXIV, Iss. 2. – P. 18–22.
Low temperature extraction of essential oil bearing plants by liquificate gases. 2. Flowers of linden (Tilia tomentosa Moench.) [Electronic resource] / Т. Atanasova, N. Nenov, Т. Girova et al. – 2009. – Available at : http://www.e-xtracts.com/ ExtractumScientific6.pdf.
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