India was enriched with a variety of herbs, shrubs, trees, and seeds used in the ancient Indian system of medicine and reported to possess beneficial medicinal effects in curing illness. Trigonella foenum-graecum is used in both forms in food and medicine in Asia. Seeds are eaten as sprouts and rich in secondary metabolites, potential sources of drugs, and essential oils of therapeutic importance. Due to their bitter taste, seeds are a good source of resin, protein, fibre, and mucilage. The critical advantages of medicinal plants' therapeutic uses in various ailments are their safety besides being economical, effective, and easy availability [1,2,3]. Our ayurvedic system praised this herb and documented its healing capacity in its ancient texts.
Among the plants known for medicinal value in Indian medicine, the plants of genus Trigonella foenum-graecum Linn belong to the family Fabaceae are vital for their therapeutic potentials. It is extensively cultivated as a semi-arid crop in northern Africa, the Mediterranean, India, and Canada [4]. Trigonella foenum-graecum (Fig. 1) grows as an erect annual with long, slender stems reaching 30–60 cm in height. The plant bears grey-green, tripartite, toothed leaves. White or pale-yellow flowers appear in summer and develop into long, slender, sword-shaped seed pods with a curved, beak-like tip. It has been reported that Trigonella foenum-graecum could be employed in the medicinal, pharmaceutical, and nutraceutical fields [5]. It is used as an aphrodisiac, astringent, demulcent, carminative, stomachic, diuretic, emmenagogue, emollient, expectorant, galactagogue, restorative, and tonic [3]. Trigonella foenum-graecum is used to treat digestive issues, bronchitis, TB, fevers, sore throats, wounds, arthritis, abscesses, swollen glands, skin irritations, diabetes, loss of appetite, ulcers, and menopausal symptoms, as well as cancer. Antidiabetic, antifertility, anticancer, antibacterial, and antiparasitic effects have been described for Trigonella foenum-graecum [3, 6]. An infusion of the leaves is used as a gargle for recurring mouth ulcers in traditional medicine. It is used as an emollient in poultices for boils, cysts, and other skin irritations. It is used to control blood pressure and lower blood sugar levels. Trigonella foenum-graecum has been shown to reduce inflammation, alleviate congestion, and fight infection. It loosens and eliminates excess mucus and phlegm, as well as relieving sinus and lung congestion. The Trigonella foenum-graecum seed is used in Chinese medicine to treat abdominal discomfort, chilblains, cholecystitis, fever, hernia, impotence, hypogastric, nephrosis, and rheumatism [7]. Trigonella foenum-graecum is known to contain alkaloids, flavonoids, salicylate, and nicotinic acid. The chemical composition of Trigonella foenum-graecum (like seeds, husk, and cotyledons) showed that endosperm had the highest saponins (4.63 g/100 g) and protein (43.8 g/100 g) content [6, 8]. T. foenum-graecum contains 45–60% galactomannans, 20–30% proteins high in lysine tryptophan, 5–10% lipids, pyridine alkaloids, trigonelline (0.2–0.38%), choline (0.5%), carpaine gentianine, flavonoids luteolin, apigenin, quercetin, orientin, isovitexin vitexin, amino as 4-hydroxyisoleucine (0.09%), histidine, arginine lysine, calcium, saponins, glycosides steroidal sapogenins on hydrolysis (yamogenin, Diosgenin, neotigogenin, tigogenin), sitosterol, cholesterol, vitamin A, B1, C, and nicotinic acid [9,10,11]. Trigonella foenum-graecum seeds mainly contain Diosgenin [(3β,25R)-spirost-5-en3-ol], a plant-derived steroid sapogenin [3, 12]. Diosgenin [(3β,25R)-spirost-5-en3-ol] (Fig. 2), a phytosteroid sapogenin, possesses several biological activities, including anticancer, hypolipidemic, anti-inflammatory, and antidiabetic ones. It also plays a beneficial role in the cardiovascular system and reduces bone loss in osteoporosis [13]. Some reported literature quantifies Diosgenin in Trigonella foenum-graecum seeds by chromatographic methods specially HPLC [14,15,16,17,18,19]. In the current study, Diosgenin was selected for qualitative and quantitative estimation in the coded formulation and Trigonella foenum-graecum seeds by HPTLC and HPLC. The aim of the present study is to develop quality control protocols for the coded ayurvedic drug by analysing the reference standard of bioactive compounds present in the formulation and respective ingredient—evaluation of HPTLC fingerprint profiling along with dominant biomarkers and estimation of bioactive markers Diosgenin by HPLC.