INTRODUCTION:

Plants of the genus Tephrosia belong to the family Fabaceae (Leguminosae) and the subfamily Papilionaceae with more than 350 species approximately, that are mainly distributed in tropical, subtropical and arid regions of the world^1-3. They are erect herbs or soft or woody shrubs with dense foliage grown to a height ranging from 0.5 to 4m. Leaves are compound, leaflets are inverted lance-shaped or obovate, 7–15cm long and 0.3–1cm wide. Flowers are pea shape, white, purple or pinkish, 7mm long, in a few-flowered, leaf-opposed, raceme-like clusters. Plants are self-pollinated and produce linear-long pods, 2.5–4.0cm long and 3–4mm wide. Seeds are ellipsoid, dark brown⁴. Many plants from this genus have been used traditionally for the treatment of diseases like inflammatory diseases, syphilis, diarrhea, asthma, respiratory disorders etc⁵. Tephrosia purpurea, an important plant of the genus is used in the treatment of cancers, inflammatory diseases and dysentery in the commune of Badaguichiri in Niger⁶. Phytochemical investigations revealed the presence of a number of phytoconstituents such as flavonoids, isoflavonoids, sterols, triterpenes. The bioactivity associated with the plant has been studied extensively, indicating the phytoconstituents present in the Tephrosia genus manifested various biological activities such as anti-diabetic, anti-ulcer, anti- diarrheal, wound healing, anti-inflammatory, insecticidal, anti-viral, anti-protozoal, anti-fungal, anti-plasmodial, and many other activities^7. Analysis of literature data provided no information on pharmacognostic studies conducted on Tephrosia species collected in Mali. It is in this context that the present study was initiated and aimed to determine the pharmacognostic parameters of the leafy stems of three Tephrosia species easily accessible and frequently available in Mali in order to contribute to the writing of their monographs. These species of Tephrosia are Tephrosia bracteolata Guill. & Perr., Tephrosia mossiensis A. Chev. and Tephrosia linearis (Willd.) Pers.

MATERIAL AND METHODS:

Plant Material:

It consisted of the leafy stems of Tephrosia bracteolata, Tephrosia mossiensis and Tephrosia linearis, harvested in September 2023 on the hill of Sangarébougou, Bamako, Mali. The harvested samples were dried under shade for three weeks in the drying room of the National Institute for Research on Traditional Medicine and Pharmacopoeia (known as INRMPT from its French name). Once dried, they were pulverized using a mill (FORPLEX GRINDER). The botanical identity of the plants was confirmed by Seydou Mamadou DEMBELE, Water and Forestry Engineer, head of the Ethnobotany and Raw Materials Department of INRMPT Voucher specimen is deposited in the Herbarium of the same institution under the number 0831 for Tephrosia bracteolata, 0835 for Tephrosia mossiensis and 0312 for Tephrosia linearis.

Determination of Botanical Characters:

a) Macroscopic studies:

The macroscopic study is a morphological study. This study allowed us to describe leafy stem from the three species of Tephrosia⁸.

b) Determination of organoleptic characters:

The organoleptic characters of the powders of the three Tephrosia species were determined using the method described by Fofie et al.⁹. The organoleptic characters determined concerned the color, taste, smell and texture of the powder.

c) Microscopical examination :

Micrographs of the powders of the three species of Tephrosia were carried out according to the method described by Fofie et al.⁸. The powders of the three Tephrosia species were mounted between slide and coverslip in a 5% diluted aqueous solution of potassium hydroxide (KOH). The microscopic elements were observed with the objective 40 of an OPTIKA ITALY microscope and then photographed using an iPad device that was directly connected to the microscope.

Physicochemical Analyses:

Physicochemical parameters such as moisture content, ash values (total ash and ash insoluble in 10% dilute hydrochloric acid), and extractive values for water and 70% ethanol were determined according to methods described in the European Pharmacopoeia^10-12.

Preparation of Extracts:

a) Decoction: Five grams of powder from each sample were boiled in 50mL of distilled water for 15 minutes, then filtered through cotton and gauze pads.

b) Infusion: 50mL of boiling distilled water was added to five grams of powder from each sample, then allowed to stand for 15minutes. The mixtures were filtered through cotton and gauze pads.

c) Maceration: 50mL of a 70% diluted hydroethanolic solution (30v/70v) was added to five grams of powder from each sample, then allowed to stand for 24hours. The mixtures were filtered through cotton and gauze pads.

The filtrates were concentrated using a rotary evaporator and then dried at 60°C for 24hours.

Characterization of chemical and anti-radical constituents:

a) Characterization of chemical constituents:

The main chemical groups were characterized by tube reactions using conventional reagents. The screened chemical groups were alkaloids (Dragendorff reagent), anthocyanins (acid and alkali reaction), hydroxyanthracene derivatives (Bornträger reaction), flavonoids (Cyanidine reaction), saponins (foaming test), tannins (trichloride reaction), and sterols and triterpenes (Lieberman reaction)^13,14. The results were categorized as follows:

Strongly positive reaction:+++; Moderately positive reaction:++; Weakly positive reaction:+; Negative reaction:

b) Characterization of anti-radical constituents:

The anti-radical constituents of the extracts were characterized after thin layer chromatography using the 2,2-diphenyl-1 picrylhydrazyl (DPPH) radical reduction test¹⁵. Extracts from infusion, decoction and maceration were deposited on an aluminum plate covered with silica gel type G₆₀F₂₅₄and then developed in the solvent system Butanol – Acetic Acid – Water (60 – 15 – 25). The chromatograms of the extracts were revealed with a methanolic solution of DPPH (2mg/mL). The antioxidant constituents appear as yellow spots on a purple background.^16,17.

RESULTS AND DISCUSSION:

Botanical characters:

a) Morphological Characteristics:

The stem of T. bracteolata is erect, cylindrical and full, solitary, very branched, 0.50 to 2.50m high. It is covered with fine longitudinal wrinkles. It is covered with short, applied, silvery pubescence. The leaves are compound and alternate. They are imparipinnate, comprising 5 to 15 pairs of lateral leaflets and one terminal leaflet. The rachis is 8 to 30cm long. The petiole is 1 to 5cm long. The leaflets are linear to oblong, with a rounded, mucronate apex, a broad wedge-shaped base, and an entire margin. They increase in length towards the apex of the rachis and measure 4.5 to 8cm long and 3 to 5mm wide. They are borne on a 1 to 2mm long petiole. The upper surface is glabrous, marked with numerous oblique and parallel secondary veins, while the lower surface is covered with a silvery pubescence consisting of stiff, applied hairs (Fig. 1A). The stem of T. mossiensis is generally erect, thin and pubescent, with a height of up to 1.5m. It is covered with a fine down called "indumentum". The leaves of Tephrosia mossiensis are compound and alternate. They are imparipinnate comprising 4 to 7 pairs of opposite leaflets that decrease in size towards the base. The rachis is 5 to 7cm long. The leaflets are obovate or elliptical in shape, measuring 15 to 30mm long and 5 to 15mm wide. Both surfaces of the leaflets have a velvety and pubescent texture (fig.1B). The stem of T. linearis is slender, erect, cylindrical, and full. It is covered with fine longitudinal wrinkles. It is covered with short, appressed pubescence. The leaves are compound and alternate. They are imparipinnate, comprising 4 to 9 pairs of lateral leaflets and one terminal leaflet. The rachis is 3 to 5cm long. The petiole is 3 to 10mm long. The leaflets are linear to oblong, with a rounded, mucronate apex, an acutely wedged base, and an entire margin. They increase in length toward the apex of the rachis and are 15 to 40mm long and 2 to 4mm wide. They are subsessile. The upper surface is glabrous, marked with pinnate secondary veins, while the lower surface is covered with silvery pubescence consisting of stiff, appressed hairs (fig. 1C). These data will contribute to the identification of the leafy stems of the three species before they are powdered.

Figure 1: Image of the three Tephrosia. A-T. bracteolata ; B-T. mossiensis ; C-T. Linearis

b) Organoleptic Characteristics and Microscopic Elements of Powders:

The powders of the leafy stems from the three Tephrosia species had a lichen-green color, an uncharacteristic odor, an insipid flavor, and a semi-coarse texture. Microscopic examination of the powders revealed the presence of epidermal fragments with anomocytic stomata, palisade parenchyma, fiber groups, spiral xylem, and punctate xylem in the powder samples of the three Tephrosia species (Figure 2). The differential characteristic microscopic elements from the powdered samples of the three species were identified as multicellular covering trichomes in the T. linearis, straight unicellular covering trichomes for T. bracteolata, slightly curved unicellular covering trichomes and starch grains in the T. mossiensis (Figure 3). Organoleptic characteristics and microscopic elements are important elements in the distinction of a plant material, as they play a role in the detection of falsified or substituted plant material⁸. These information on the organoleptic characteristics and microscopic elements will contribute to the botanical quality control of future samples of powders from the leafy stems of the three species of Tephrosia.

Figure 2: Common microscopic elements of the powder of the three Tephrosia. GX40

Figure 3: Differential microscopic elements of the powder of the three Tephrosia. GX40

A: T. Bracteolata powder; B: T. linearis powder; C: T. mossiensis powder

Physicochemical Parameters:

Physicochemical analysis is an essential element of quality control of plant material. The results of the physicochemical analysis are presented in Table I. The moisture contents in the powders from the leafy stems of the three species of Tephrosia analyzed were less than 10%, which would minimize the risk of proliferation of bacteria and fungi (yeasts and molds) in these powders during their conservation¹⁸. Total ash contents were between 4-5% (see Table I). Total ash contents were significantly higher in the leafy stem powder samples of T. bracteolata and T. mossiensis compared to that of Tephrosia linearis. These results suggest that the powdered leafy stem of T. bracteolata and T. mossiensis would be richer in inorganic minerals than the powdered leafy stem of T. linearis¹⁸. The acid (10% HCl) insoluble ash contents were less than 0.5% in all three samples (see Table I). These results suggest that powdered samples of the three species have a very low proportion of siliceous elements such as dust and sand¹⁸. In Table I the 70% ethanol-soluble extractive values for each sample were higher than those of water-soluble extractive values. These results suggest that 70% hydroethanolic solution is the best solvent to extract phytochemicals from these three Tephrosia species. So these current findings are similar to those found by Rajabudeen et al. who showed that alcohol is better than water to extract the phytoconstituents of Tephrosia villosa (L.) Pers¹⁹.

Table 1: Moisture and ash contents of the leafy stems of the three species of Tephrosia

Physicochemical parameters	*T. bracteolata*	*T. mossiensis*	*T. linearis*
Moisture content (%)	5.5 ± 0	5.37±0,75	5± 0,40
Total ash content (%)	5 ± 0.40	4.62±0,25	4±0
Acid insoluble ash content (%)	0.37 ± 0,17	0.25±0	0.25±0
Water-soluble extractive values (%)	8.06 ± 0.03	7.34 ± 0.54	6.32± 0.74
70% ethanol-soluble extractive values (%)	8.58 ± 0.08	8.74 ± 0,37	9.08 ± 0.34

Chemical and anti-Radical Constituents:

a) Chemical Constituents:

Phytochemical screening shows that the leafy stems of the three Tephrosia species have a very similar phytochemical composition with a slight difference (Table II). Flavonoids, leucoanthocyanins, carbohydrates, sterols and triterpenes and tannins were identified in the powders of the three Tephrosia species. Anthracene glycosides were absent in the powdered samples of the leafy stems of Tephrosia bracteolata. On the other hand, saponins were found only in the powdered sample of the leafy stems of Tephrosia mossiensis. The results obtained with the powdered samples of Tephrosia bracteolata and Tephrosia linearis are slightly different from those reported in the literature. The presence of alkaloids was reported in the samples of the leafy stems of these two species collected in Nigeria^20,21, which is different from our results. This difference could be explained by several factors such as the harvesting place, the harvesting period, the growing stage of the plant. About Tephrosia mossiensis, we did not find in the literature any previous work concerning its phytochemical composition.

Table 2: Chemical groups screened in the powders of the three species of Tephrosia

Chemical groups	*T. bracteolata*	*T. mossiensis*	*T. linearis*
Alkaloids	-	-	-
Anthocyanins	-	-	-
Anthracene glycosides	-	++	+
Flavonoids	+++	++	+++
Leucoanthocyanins	+++	++	++
Carbohydrates	+++	+++	+++
Saponins	-	+	-
Sterols and triterpenes	+++	++	++
Tannins	+++	++	+++

b) Anti-radical Constituents:

The appearance of yellow spots on a purple background indicates the antioxidant capacity of the aqueous and hydroethanolic extracts of the three Tephrosia species (Figure 4). The aqueous and hydroethanolic extracts of the leafy stem samples from the three Tephrosia species have a similar antioxidant profile. However, it is necessary to determine by spectrophotometric method the effective concentration 50 (EC₅₀) of the extracts that will reduce the DPPH radical. This antioxidant activity could be due to the presence of bioactive compounds of polyphenolic types such as tannins, flavonoids and leucoanthocyanins which are known for their ability to reduce free radicals²². This antioxidant activity of extracts from the leafy stems of the three Tephrosia species could be beneficial in the management of numerous pathologies and/or symptoms such as pain, inflammation, cancers and cardiovascular diseases. However, pharmacological studies are necessary to confirm these hypotheses.

Figure 4: Chromatograms of extracts from the three Tephrosia species revealed by DPPH.

I: Infusion; D: Decoction and M: Maceration

CONCLUSION:

At the end of our study, it emerged that samples of powdered leafy stems from Tephrosia bracteolata, Tephrosia mossiensis, and Tephrosia linearis have botanical, physicochemical, and phytochemical profiles slightly different but they present a similar anti-radical profile. Data from this study contributed to monographs of the leafy stems of these plants and will be helpful in the quality control of leafy stem samples from these plant species to make Phytodrugs and also in student training.

CONFLICT OF INTEREST:

No conflict of interest.

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Received on 27.05.2025 Revised on 30.08.2025

Accepted on 03.11.2025 Published on 31.01.2026

Available online from February 07, 2026

Res. J. Pharmacognosy and Phytochem. 2026; 18(1):26-30.

DOI: 10.52711/0975-4385.2026.00005

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