Anti-Inflammatory Subfractions Separated from Acidified Chloroform Fraction of Fenugreek Seeds (Trigonella foenum-graecum L.)

ABSTRACT

Considering the side effects of current anti-inflammatory drugs, novel therapeutic agents are desired. We have succeeded in separating flavonoid-rich fractions with anti-inflammatory effect from fenugreek seeds (Trigonella foenum-graecum L.). In this work, we aimed to carry out further fractionation to find active anti-inflammatory subfractions. Trigonelline content of the plant was determined by spectrophotometric method. Fenugreek seeds were extracted consecutively with petroleum ether, acidified chloroform (ACC), alkaline chloroform (AKC), methanol, and water. ACC fraction, which had exhibited the highest anti-inflammatory effect, was further fractionated using column chromatography. Obtained subfractions were evaluated using carrageenan-induced paw edema (CIPE) method. Animals were pretreated by test compounds, and after 30 minutes edema was induced by subcutaneous injection of 100 µl of 1% w/v carrageenan into the right paw of animals. Volume difference of both paws was measured at different times after carrageenan injection. The concentration of trigonelline was determined as 16.2%. ACC fraction inhibited paw edema significantly in comparison to control (p < .05). Four subfractions (dry weight percentage basis) were selected for pharmacological study. F3 subfraction exhibited the greatest inhibition at 15 mg/kg (p < .001). ACC fraction and F4 significantly inhibited paw edema at doses of 5, 10, and 15 mg/kg (p < .001). Phytochemical studies indicated the presence of flavonoids in ACC and active subfractions. Further separation can lead to finding active components from active subfractions, which probably belong to flavonoid phytochemicals. Considering the gastroprotective effect of fenugreek, we hope the separated fractions also would be free of gastrointestinal side effects.

KEYWORDS:

• carrageenan
• fenugreek
• flavonoid
• subfraction

Declaration of interest

The authors declare no conflicts of interest. The authors alone are responsible for the content and writing of the article.

Funding

The authors greatly appreciate the vice chancellor of research for financial support of the project.

About the authors

Mostafa Pournamdari, PhD of analytical chemistry. Pharmaceutics research Center, Institute of Neuropharmacology, interested in the chemistry of natural products.

Ali Mandegary, PhD of toxicology, Neroscience Research Center, interested in neuropharmacology.

Fariba Sharififar, PhD in Pharmacognosy, Herbal and Traditional Medicines Research Center, interested in medicinal plants , pharmacology and bioactive separation and ethnobotany.

Ghazale Zarei, PharmD, Herbal and Traditional Medicines Research Center, interested in medicinal plants.

Rahele Zareshahi, PhD, student in traditional pharmacy, Herbal and Traditional Medicines Research Center.

Amir Asadi, BS, Herbal and Traditional Medicines Research Center, interested in medicinal pants and ethnobotany.

Mohammad Mehdipour, MS, Neuroscience Research Center, interested in toxicology and pharmacology of medicinal plants