Development of Methods for Quantitative Determination of The New Antioxidant Dodecyl(3,5-Dimethyl-4-Hydroxybenzyl)Sulfide

The prevention and the therapy of such diseases as atherosclerosis, hepatitis, and cancer is nowadays one of the most significant medical problems. Dodecyl(3,5-dimethyl-4-hydroxybenzyl)sulfide was synthesized at the chair for chemistry of Novosibirsk State Pedagogical University. This substance is a new promising phenolic antioxidant, which can be used in the complex therapy of abovementioned diseases. The aim of this study was to develop methods for quantification of dodecyl(3,5-dimethyl-4-hydroxybenzyl)sulfide for standardization and bioanalytical studies. The mercurimetric titration, HPLC-UV and voltammetry were used. The methods validation was performed according to general pharmacopoeial monograph OFS.1.1.0012.15 "Analytical methods validation" and OFS.1.1.0013.15 "Statistical processing of chemical experiment results" of the Russian State Pharmacopoeia XIV. All the results meet the acceptance criteria in parameters specificity, linearity, precision and accuracy. The mercurimetric titration of dodecyl(3,5-dimethyl-4-hydroxybenzyl)sulfide is recommended for including in chapter "Assay" of the original monograph project for substance. The HPLC-UV method may be useful for the determination of the object of study in dosage forms. The voltammetric method is promising for bioanalytical researches.

Fruits of Crataegus submollis Sarg.: morphological and anatomical study

Actuality. Fruit of Crataegus L. are successfully used in medicine in the Russian Federation as a source of cardiotonic substances. One of the sources of raw material is Crataegus submollis Sarg., which is cultivated as an edible and ornamental plant everywhere. Previous studies revealed that preparations from fruit of Crataegus submollis Sarg. show diuretic activity and antidepressant action. Therefore, we consider Crataegus submollis Sarg. to be of interest as a new source of medicinal plant raw material, its fruit can be used both fresh and dried. Purpose. The aim of the article is to study the morphological and anatomical features of fresh fruit of Crataegus submollis Sarg. Materials and methods. The material of the study was the fresh and frozen fruit of Crataegus submollis Sarg. collected in September October 2018 in the Botanical garden of Samara University (Samara). The research was carried out by the method of light microscopy in transmitted and reflected light in the light background. Results. Performed analysis allowed to reveal structural characteristics of fruit of Crataegus submollis Sarg. which included cellular structure of the epidermis, lack of stomatal apparatus in epidermis, presence of cortical pores, trichome characteristics, and seed constitution. Conclusion. Our findings allow to develop the section Microscopic Signs of the pharmacopoeial monograph Fresh Fruit of Crataegus submollis Sarg..

THE new aspects iN MoRPHoloGICAL AND anatomical DiAGNosTics of the herb monarda fistulosa l

Topicality. The herb of Monarda fistulosa L. cultivated in the Russian Federation is one of the promising sources of essential oils, flavonoids and other biologically active compounds. The problem of standardization of raw materials of this plant is not fully resolved. Objective. The article is aimed at specification and addition of anatomical and histological signs of leaves and stems of Monarda fistulosa, as well as at the study of the luminescent features of their tissues. Materials and methods. The material of the study was the stems and leaves of the Monarda fistulosa, collected in the flowering period in July 2017 in the Botanical Garden of Samara University. The experiment was carried out by the method of light microscopy in transmitted and reflected light on a light field in accordance with the requirements of the monograph of the State Pharmacopoeia of Russia, XIV edition. Results. The article presents the results of a morphological and anatomical study of the herb of Monarda fistulosa. Additionally, the diagnostic features of the studied plant were determined, and the luminescence features of the tissues of the stems and leaves were also described. Conclusion. The obtained data allow us to develop a section “Microscopic signs” of the Pharmacopoeial monograph for the new type of medicine plant material - “herb of Monardae fistulosae”.

Comparative Study of the Biologically Active Substances Composition and Content in Meadowsweet (Filipendula ulmaria (L.) Maxim) Crude Herbal Drugs (Herb, Leafs, Flowers) of Russian Origin

Filipendula ulmaria (L.) Maxim. (Meadowsweet) is known in traditional medicine as anti-inflammatory, wound-healing,astringent and antibacterial remedy. However recent studies show that it also has neurotropic activity. In Russia meadowsweet flowers are used as crude herbal drugs (temporary pharmacopoeial monograph 42-1777-87), also leafs and herb are used in the traditional medicine. Objective of the study was to carry out comparative investigation of composition and content of major biologically active compounds (BAC) in Filipendula ulmaria herb, flowers and leafs by thin-layer chromatography, differential spectrophotometry with aluminum chloride reagent (total flavonoids in terms of rutoside), gravimetry (total extractives, extracted by water), permanganatometric titration (total tannins in terms of tannin). Rutoside, tannin, gallic acid and salicylic acid were identified in Filipendula ulmaria herb, flowers and leafs by TLC. Also we analyzed content of substances extracted by water, flavonoids and tannins. Total extractives, extracted by water in F. ulmaria herb is 13.12±0.10%, in leafs – 13.98±0.37%, in flowers – 18.09±0.17%. Total tannins in F. ulmaria herb is 11.87±0.47%, in leafs – 12.06±0.18%, in flowers – 12.26±0.29%. Total flavonoids in F. ulmaria herb 4.34±0.17%, in leafs – 6.98±0.23%, in flowers – 11.75±0.57%. The obtained data will be used for development of a pharmacopoeial monograph project "Filipendula ulmaria (L.) Maxim., herba" for inclusion in the State Pharmacopoeia of the Russian Federation.

STANDARDIZATION OF SNOWDROP (GALANTHUS L.) HERBAL PHARMACEUTICAL SUBSTANCES BY UV-SPECTROPHOTOMETRY

Objective: The objective of the present study is to evaluate the electronic ultraviolet (UV) absorption spectra of herbal pharmaceutical substances (70% ethanol extracts) prepared from medicinal plant material of two snowdrop species – Galanthus woronowii Losinsk and Galanthus nivalis L.Methods: The groups of biologically active substances were investigated by UV-spectrophotometry including special sample preparation for flavonoids and alkaloids.Results: In the present study, data were obtained characterizing the composition of the main biologically active compounds of the genus Galanthus L., received its “spectrophotometric profiles.” On the basis of spectral characteristics of G. woronowii and G. nivalis flavonoids and alkaloids, we confirm feasibility for using galantamine, lycorine, quercetin, and its glycosides (hyperoside et al.) state standard samples in standardization analysis.Conclusion: It is shown that this method can be used to assess the quality of herbal pharmaceutical substances, serve to prove their identity in pharmacopoeial monograph.

MICROSCOPIC DIAGNOSTIC FEATURES OF WHOLE, MILLED AND POWDERED PLANT MATERIAL “RHIZOMES OF COMMON TURMERIC”

Aim - the determination of microscopic diagnostic features of rhizomes of common turmeric (Curcuma longa L.), ginger family (Zingiberaceae). Materials and methods. We have studied whole, milled and powdered plant material of turmeric from different manufactures. The study was carried out with the use of the digital microscopes “Motic DM111” (Korea) and “Motic DM-39C-N9GO-A” (Korea). Results. It was determined that the microscopic diagnostic features of rhizomes of turmeric include the presence of cells with sinuous walls and structured content in the form of colored folding in the parenchyma; closed collateral vascular bundles lie scattered in the parenchyma, their conductive elements have narrow lumen. Vascular bundles contain pigment cells considerably smaller in comparison with the size of vessels. Pigment cells with drops of orange-red essential oil are also found in the parenchyma. In milled and powdered plant material, the fragments of all these elements are observed. In the fragments of powdered rhizomes of turmeric, numerous cells with yellow content are found; the parenchyma cells have less expressed folded structure. Fragments of vessels are found both in milled and powdered material. Conclusion. The results of this study were included in the section “Microscopy” of the project of pharmacopoeial monograph “Curcuma longa L. rhizomes”.

Development of chromatographic methods for thebaine detection and quantification along with some of related alkaloid derivatives

<p class="Body">Thebaine is naturally occurring alkaloid which is of great importance in the pharmaceutical industry and up to now there is not a Pharmacopoeial monograph for this compound. There is a definite need for development of simple procedures for routine quality assessment based on physical and spectroscopic properties (melting point, UV-Vis, IR, specific optical rotation) and even more importantly a need for development of chromatographic methods for determination of purity.The main objective of this work was to develop RP-HPLC-UVD method for quantification of thebaine and also to develop complementary GC-MS method for qualitative analysis. The developed HPLC method utilized RP-C18 monolithic column, gradient elution with trifluoroacetic acid and formic acid in water and in acetonitrile and detection at 285nm. The effects of the relevant chromatographic conditions was investigated and the optimized and validated method was usеd for analysis of three batches of thebaine. </p><p class="Body">A procedure that contained different methods for determination and identification of thebaine, was performed with main focus on the HPLC method suitable for quantitative and qualitative determination. In parallel, to complete the purity profile, the three batches of thebaine were analyzed by IR, UV-Vis spectroscopy and specific optical rotation and melting point determination. The analysis of thebaine can be performed in few steps by simple and reproducible methods like IR spectroscopy, specific optical rotation, melting point and the developed HPLC method, later proved by its validation. The developed HPLC-UV method is suitable for transfer to HPLC-MS method. The GC-MS method is quite suited for identification of thebaine and of the impurities, but is limited to free bases. Additionally, pseduomorphine and <em>N</em>-oxide derivatives of the related alkaloids are not suitable for GC-MS analysis.</p>

Providing Some Pharmacopoeial Standards for Nigerian Allium cepa Var. cepa Linn

The essence of standardization of medicinal plants is to produce herbal medicines with acceptable safety, efficacy, quality and reproducibility. Allium cepa var. cepa L. (Amaryllidaceae) bulbs purchased from Sasha Market, Ibadan, Nigeria during the rainy season was subjected to organoleptic, morphological and physico-chemical experiments. The total ash gave 5.40 ± 0.001%, acid-insoluble ash (0.62 ± 0.0001%), water-soluble extractive (2.30 ± 0.004%), alcohol-soluble extractive (1.14 ± 0.005%) and moisture content (91.1 ± 0.009%). Microscopical evaluation of the bulb showed distinct cell-walls devoid of nucleus while the inner epidermis possessed well delineated cell-walls, transparent cytoplasm containing nucleus but without stomata. Histochemical experiments showed lignified, suberized and cellulosic cell-walls, starch grains, oils, resins, tannins and anthraquinones. The study has provided some data for the pharmacopoeial monograph on Allium cepa var. cepa bulb as a possible antidiabetic agent for inclusion into the 2nd editions of the Nigerian Herbal Pharmacopoeial (NHP) and West African Herbal Pharmacopoeial (WAHP).Keywords: Allium cepa var. cepa, Pharmacopoeial, Standardization, Monograph

Determination of an infectious dose of Leptospira for the performance of challenge test in assessing the efficacy of Leptospira vaccines

The efficacy of vaccines against leptospiral disease can be determined objectively by challenge test in experimental animals. Selection of suitable leptospiral challenge strains and determination of an optimal challenge dose to prove exactly that the given vaccine Leptospira serotype induces protective immunity in vaccinated dogs is a critical point in performing challenge experiments. The aim of our study was to verify and determine an appropriate challenge dose for efficacy tests in dogs for the following Leptospira serovars: L. grippotyphosa, L. icterohaemorrhagiae and L. canicola. The appropriate challenge dose was determined on the basis of pathognomonic symptoms of infection, Leptospira capture at cultivation and pathological changes in dogs infected experimentally with various doses (5 × 104 - 5 × 108) of Leptospira serovars. A dose of 5 × 106 of each respective serovar administered intraperitoneally was determined to be a suitable challenge dose. The dogs infected with the selected dose showed the typical symptoms of the disease and met the requirements of an objective and standard evaluation of the vaccine efficacy according to the pharmacopoeial monograph. A study of such extent was done for the first time in the Czech Republic.