Pharmacognostic analysis of medicinal plant material with the help of Altami Studio software and Altami BIO 2 microscope
Pharmacognosy is a complex scientific discipline that studies questions about medicinal plants (MP), medicinal plant raw (MPR) and medicinal raw of animal origin. Pharmacognosy – one of the oldest branches of pharmacy, is a core discipline that forms a pharmacist model in accordance with the educational standard “Pharmacy” (vocational education – specialty).
About 18,000 medicines (MP) are used in the medical practice of the Russian Federation, among which approximately 40% is produced from MPR. Nowadays the use of MPR for treatment and prevention of a range of diseases tends to increase, and according to WHO forecasts, in the near future (15-20 years) the proportion of MP on the basis of MPR may exceed 50%.
Today pharmacognosy becomes particularly relevant as with the beginning of the XXI century there are qualitative transformations in terms of studying MP and MPR. This became possible due to the fact that this science began to use modern methods in the research of plant objects, among which digital optical microscopy (DOM) holds an honored position.
In 2015 the XIII edition published in the State Pharmacopoeia of the Russian Federation reflected the possibilities of modern equipment, as well as new requirements for regulatory documentation (RD). In this regard, the DOM method, which makes it possible to identify and document (in the form of microphotographs) the main anatomical and diagnostic signs of MPR, is of particular importance.
The principle of DOM is based on the analysis of the image obtained from a digital camera installed on a microscope, using specialized software. The main instrument is, of course, a light (optical) microscope, which is used in everyday laboratory tests to assess the authenticity of the MPR in accordance with the “Microscopy” section of the current regulatory documentation (private pharmacopoeial articles on the MPR). In recent years a new part appeared – a digital complex consisting of a photo / video camera and a personal computer with special software installed. Being the main professional equipment for the DOM, the digital camera is responsible for outputting the image and it provides the proper quality of digital photographs.
The DOM method along with other pharmacognostic methods of research serves as a methodological basis for improving approaches to the standardization of MPR and MP based on MRP. In order to confirm the authenticity of both whole and cut, powdered MRP pharmacopoeial microscopic analysis (one of the types of pharmacognostic analysis) based on the identification of anatomical and diagnostic features is used.
Not only at the stage of RD development, but also in everyday routine analysis,the application and widespread introduction of the DOM method for microscopic analysis, seems very relevant for pharmaceutical practice.
Materials and methods
Objects of the study: samples of drooping birch (Betula pendula Roth), hazel (Corylus avellana L.), eller (Alnus glutinosa L.), goat willow (Salix caprea L.), cut-leaved maple (Acer negundo L.); collected in forest park areas of Moscow and the Moscow region ; the grass of anisic giant hyssop (Lophanthus anisatus Benth.) of cultivar “Astrakhan 101” of the VNIIOB GNU selection ; flowering plants of Voronov’s snowdrop (Galanthus woronowii Losinsk.) and snowdrop (Galanthus nivalis L.), harvested in the Botanical Garden of the First I.M. Sechenov Moscow State Medical University.
Investigation of microscopic signs of LRS was carried out in accordance with the requirements of general pharmacopoeial articles of the GF XI-XIII ed .: “Roots, rhizomes, bulbs, tubers, corms” (OFS.1.5.1.0006.15); “Herbs” (OFS.1.5.1.0002.15); “Leaves” (OFS.1.5.1.0003.15); “Flowers” (OFS.1.5.1.0004.15); “Technique of microscopic and microchemical study of medicinal plant material and medicinal herbal preparations” (OFS.1.5.3.0003.15). While conducting microscopic analysis using the method of CMI, the studies and photographs were performed on a microscope “Altami BIO 2” (eyepiece 10 × and lenses: 4 ×, 10 ×, 40 ×, 100 ×) with the help of a digital eyepiece camera UCMOS05100KPA; The images were processed with Altami Studio software.
Results and discussion
The main anatomical and diagnostic features were identified, during the study of drooping birch (Betula pendula Roth) and impurity species (pic. 1). In the paper the results of the analysis are presented in more detail. It has been shown that morphology of the pollen grains as a diagnostic feature can be used in microscopic examination. Morphological and metrological characteristics of the investigated samples of drooping birch and some of the most frequently encountered impurity species of woody plants are hazel (Corylus avellana L.), eller (Alnus glutinosa L.), goat willow (Salix caprea L.), cut-leaved maple or maple ash (Acer negundo L.) are shown. The use of two techniques for the preparation of temporary microslides (including liquid – lactophenol, without the inclusion of a liquid “pollen in the air”) is proposed to determine the authenticity of the raw material. Thanks to the conducted researches, it will be possible to create modern regulatory documentation which allows to determine the authenticity and good quality of the native material used for the production of medical immunobiological slides of drooping birch in the Russian Federation.
During the analysis of anisic giant hyssop “Astrakhan 101” the following anatomical and diagnostic signs were determined: the structure of the upper and lower epidermis of the leaf from the surface (in particular, the absence of stomata from the upper side of the leaf, the presence of essential aromatic oil glands (picture 2), the epidermis of the petals, sepals, pollen grains. Metrological characteristics of the investigated organ structures are established. The differences in the sizes of the same structures are found in different parts of the same organ. An exhaustive description of microscopic features is given which is based on the study of samples of the raw material of the anisic giant hyssop “Astrakhan 101” and can be used to create RD regulating the quality of MRP.
“Aromatic oil gland, zoom 1000×.
In studying microscopic anatomical and diagnostic signs of snowdrops (G. woronowii and G.nivalis), characteristic features of the structure of the upper and lower epidermis of leaf, epidermis of peduncle, epidermis of petals from the inner and outer sides, upper and lower epidermis of covering scales, upper and lower epidermis storing scales (Picture 3) structure of roots, cell sizes and cell inclusions (calcium oxalate raffide, starch grains). The following features are typical for two species of snowdrops: tetratite type of stomatal complex, rectangular epidermal cells of leaf, epidermal cells of floral envelope with papillary outgrowths, presence in all organs except for the flower of cells with calcium oxalate raffides. The main results are presented in the paper.
During these complex studies the potential of the DOM method use in the analysis of medicinal plant raw of various morphological groups: flowers, leaves, herbs, bulbs, roots, dried and fresh, has been demonstrated. Certainly, further research will continue on the study of microscopic anatomical and diagnostic features of medicinal plants, the development of new and improvement of old regulatory documentation.