LC-ESI-MS/MS analysis, toxicity and anti-anaemic activity of Rubia tinctorum L. aqueous extract

The present study investigated the chemical profile, toxicity, and anti-anaemic activity of Rubia tinctorum root aqueous extract against phenylhydrazine induced hemolytic anaemia. Phenolic compounds were analyzed by LC-ESI-MS/MS; acute toxicity test was evaluated by administering a single dose of 2,000 mg.kg-1 of the extract; anaemia was induced by administration of 40 mg phenylhydrazine by intraperitoneal injection for 2 days. Moreover, the anti-anaemic activity was evaluated by measuring the haematological parameters of rats treated with iron and aqueous extract for 15 days. The LC-ESI-MS/MS analysis results revealed the presence of 31 phytochemical compounds, among them, citric acid was found as the most abundant. No signs of toxicity or death were recorded, indicating that the LD50 of R. tinctorum root extract is higher than 2,000 mg.kg-1. Furthermore, the aqueous extract increased red blood cell levels by 69.82 and 71.67 % in the groups treated with 200 and 400 mg.kg-1 of the extract, respectively. Besides, a significant increase in hemoglobin from 12.05 ± 0.15 to 12.9 ± 0.52 g.dL-1 was noted in rats treated with 400 mg.kg-1 of extract. Thus, the data indicate that the root extract could be considered a natural source for the treatment of anaemia.

The First Ten Years with Edirne Red

The Eurasian Journal of Family Medicine (EJFM) is completing its 10th volume with this issue as one of the first products of the dreams of a group of academicians devoted to family medicine. Eurasian Society of Family Medicine (ESFAM) has been established in 2009 with the aim of carrying out scientific education and research activities, supporting ongoing scientific activities, and informing the society, carrying out activities for community education, and establishing a foundation to operate in this field. Shortly after its establishment, the first product of the association was organizing Trakya Family Medicine Congresses (TAHEK). At first, TAHEK was organized as a national congress in 2010 but since 2016 it became an international one with the participation of scientists from many countries (1). Next year on March 23-27, 2022 TAHEK will bring together Family Medicine academicians in Edirne for the 11th time (2). Edirne (Orestia, Orestas, Uscudama, Hadrianopolis, Edrenos, Edrenaboli, Edrene, finally Edirne) being the headquarters of ESFAM, is one of the ancient civilization centers of the world with its eight thousand years of history (3,4). After its conquest by the Turks in 1361, it was the capital of the Ottoman Empire until the conquest of Istanbul in 1453. Edirne has an important place in medical and academic history. The Complex of Sultan Bayezid II, which was opened in 1488, is one of the oldest universities in Europe with its medical faculty and hospital (5,6). In addition, the smallpox vaccine was first used in Edirne, its usage was later disseminated to Istanbul and Europe (7). Another imperial legacy that Edirne has is Edirne Red. Edirne Red was a natural dye obtained from the Rubia Tinctorum plant and very resistant to sunlight and washing, and was used in Turkish carpets and silk and cotton fabrics (8). While preparing the first issue of the Eurasian Journal of Family Medicine in 2012, we have chosen Edirne Red as the color of the cover of our journal, aiming to express our respect for the city where the association was established. Over the years, Trakya Family Medicine Congresses (TAHEK) have become one of the leading Family Medicine congresses of the Eurasian region, especially in the Balkans; It has made significant progress in becoming the meeting point of distinguished Family Medicine researchers and academicians from all over the world. During this time, our journal contributed to the dissemination of thousands of academic papers in the participating countries and increased the number of its followers. Likewise, Eurasian Journal of Family Medicine (EJFM) has enriched the international literature with many researches published over the years. It is a pride for us to express that the Eurasian Journal of Family Medicine has started to be indexed by many respected international scientific indexes. Scopus and DOAJ were added to these indexes this year. At the end of the tenth volume of our journal, we are happy to express our appreciation and gratitude to our authors, reviewers, distinguished international advisory board members, editors and all our stakeholders who shared our dreams and contributed to our products.

The possibility to enhance flavonoids production in Rubia tinctorum L. callus cultures

Production of flavonoids in madder callus culture (Rubia tinctorum L.) was dependent on culture conditions and culture media composition. The content of flavonoids increased in calli maintained on media supplemented with NAA (4 mg.l-1) or NAA:BAP (4 mg.l-1 a 1 mg.l-1) in 16 h photoperiod. Flavonoids represented 2.08 – 2.25 % of callus dry mass. The presence of Cd(NO3)2 (3.1 or 31.0 mg.l-1 concentrations) negatively influenced callus growth, but enhaced the percentage of dry mass in callus cells. During 42 days of culture an increase of cadmium accumulation and even of flavonoids has been observed. The most considerable influence of CdCl2 or Cd(NO3)2 on flavonoids content has been shown in short-term experiments after 48 h of callus culture. More distinct influence has been observed under the treatment with CdCl2 (0.005 mg.l-1) in comparison with Cd(NO3)2.

Identification of Dyes in Coptic Textiles from the Museum of Faculty of Archaeology, Cairo University

High Performance Liquid Chromatography coupled to a Diode-Array-Detector (HPLC-DAD) is used to investigate samples which were extracted from ancient Egyptian textiles (4th–5th c. AD) of the Museum of Faculty of Archaeology, Cairo University. Madder is identified in several samples. According to semi-quantitative results, which are obtained from HPLC peak areas measured at 254 nm, madder that is rich in purpurin and poor in alizarin is identified in samples which were treated (i) only with madder and (ii) with madder and either indigo/woad (Indigofera species and other/Isatis tinctoria L.) or weld (Reseda luteola L.). The madder dye used in these samples could have been originated from Rubia peregrina L. However, the possible use of Rubia tinctorum L. (or other plants of the Rubiaceae family) by the Egyptian dyers cannot be ruled out, particularly if methods were developed by the ancient dyers to affect and control the relative composition of madder dye. The HPLC peak area ratio of alizarin versus purpurin is very high (>2.2) for samples which were treated with madder (probably originated from R. tinctorum) and a tannin source. Finally, in some samples, only indigoid dyes (indigo/woad) are identified.

A SINGLE BLIND RANDOMIZED CONTROLLED TRIAL ON EFFECTIVENESS OF RUBIA TINCTORUM Q IN ANAEMIA AMONG COLLEGE GOING FEMALE STUDENTS OF SEMI URBAN AREAS IN SANGAREDDY

Anaemia is a condition of having a lower-than-normal number of red blood cells or quantity of hemoglobin. Anaemia diminishes the capacity of the blood to carry oxygen. Patients with anaemia may feel tired, fatigue easily, appear pale, develop palpitations,and become short of breath.Rubia tinctorum mother tincture is a homeopathic mother tincture that is indicated for the treatment of anaemia and undernourished conditions. Rubia tinctorum mother tincture, has been found to be effective in treating anaemia, kidney and bladder calculi.This project, help to know the efficacy of Rubia tinctorum mother tincture in treating anaemia.

Detection of Adulteration in Rubia cordifolia – A Chromatographic Approach

According to traditional medicinal texts as well as modern research, Rubia cordifolia (Desi Manjistha) stem cure various diseases of blood, skin diseases, inflammations, kidney stones, fever and various Kapha disorders. However, stems of Rubia cordifolia (Desi Manjistha) are often adulterated with stems of Rubia tinctorum (Irani Manjistha) due to morphological similarity. This adulteration is extremely harmful as R. tinctorum has Lucidin as one of the chemical constituents, which has severe genotoxic effects. The present work focuses on developing a comparative HPTLC fingerprint and GC-MS analysis of R. cordifolia and R. tinctorum, so as to detect the adulteration by R. tinctorum in R. cordifolia raw material form and in formulations. Methanol extracts of the individually powdered stems were used to develop a suitable mobile phase, such that a clear spot was obtained in TLC of R. tinctorum but not in TLC of R. cordifolia using the same mobile phase. This mobile phase was further used to develop a comparative HPTLC fingerprint of the two species. Methanol extracts of R. cordifolia and R. tinctorum were used for investigation of phytoconstituents by GC-MS analysis. The mobile phase n-butanol: ethanol (6:4) was the one which gave a clear single spot at Rf 0.89 in TLC of methanol extract of R. tinctorum but not in TLC of methanol extract of R. cordifolia, at 254nm and 366nm. Further, HPTLC analysis gave results clearly differentiating the two species using the same mobile phase. Further, GC-MS analysis of R. cordioflia revealed the presence of 9 phytoconstituents while R. tinctorum revealed the presence of 6 phytoconstituents, which are different from each other. Thus, these simple yet sophisticated techniques will be very useful for herbal industry in standardization and detection of adulteration of R. tinctorum in R. cordifolia formulations, thereby benefitting the patients who consume Manjistha formulations.

Activity of Anthracenediones and Flavoring Phenols in Hydromethanolic Extracts of Rubia tinctorum against Grapevine Phytopathogenic Fungi

In this work, the chemical composition of Rubia tinctorum root hydromethanolic extract was analyzed by GC–MS, and over 50 constituents were identified. The main phytochemicals were alizarin-related anthraquinones and flavoring phenol compounds. The antifungal activity of this extract, alone and in combination with chitosan oligomers (COS) or with stevioside, was evaluated against the pathogenic taxa Diplodia seriata, Dothiorella viticola and Neofusicoccum parvum, responsible for the so-called Botryosphaeria dieback of grapevine. In vitro mycelial growth inhibition tests showed remarkable activity for the pure extract, with EC50 and EC90 values as low as 66 and 88 μg·mL−1, respectively. Nonetheless, enhanced activity was attained upon the formation of conjugate complexes with COS or with stevioside, with synergy factors of up to 5.4 and 3.3, respectively, resulting in EC50 and EC90 values as low as 22 and 56 μg·mL−1, respectively. The conjugate with the best performance (COS–R. tinctorum extract) was then assayed ex situ on autoclaved grapevine wood against D. seriata, confirming its antifungal behavior on this plant material. Finally, the same conjugate was evaluated in greenhouse assays on grafted grapevine plants artificially inoculated with the three aforementioned fungal species, resulting in a significant reduction in the infection rate in all cases. This natural antifungal compound represents a promising alternative for developing sustainable control methods against grapevine trunk diseases.