Affiliation:
1. Department of Science and Technology, KBCNMU, Jalgaon, Maharashtra, India
2. Department of Pharmacognosy, SSPM Dr. N. J. Paulbudhe College of Pharmacy, Ahmednagar, Maharashtra, India
3. Department of Pharmacognosy, SES’S R. C. Patel Institute of Pharmaceutical Education and Research, Shirpur, Maharashtra, India
Abstract
ABSTRACT
Background:
Amla, liquorice, and fenugreek have three primary bioactive phytoconstituents: diosgenin, gallic acid, and 18β-glycyrrhetic acid. This work discusses the invention and validation of an high-performance liquid chromatography (HPLC) approach for the simultaneous detection, in line with ICH criteria, of gallic acid in amla, glycyrrhetinic acid in liquorice, and diosgenin in fenugreek.
Materials and Methods:
A new simple, exact, sensitive, and validated reversed phase (RP)-HPLC technique was developed for the measurement of diosgenin, gallic acid, 18β-glycyrrhetic acid, and acid in bulk and pharmaceutical dosage form. Acetonitrile: 0.05% ortho phosphoric acid (OPA) (95:5) for diosgenin in fenugreek extract, a rheodyne manual injector with a 20 μl capacity, and a Phenomenex Luna C18 (2) (4.6 mm × 250 mm, 5) chromatographic apparatus were used for the separation. In Amla extract, methanol: 0.05% OPA (70:30) for gallic acid liquorice extract: Methanol: Acetonitrile: 0.05% OPA for 18β-glycyrrhetic acid A solution of ophosphoric acid (OPA) was used to maintain the pH at 3.00. The detection wavelengths utilized were 205 nm for diosgenin, 272 nm for gallic acid, and 250 nm for 18β-glycyrrhetic acid. The flow rate was set at 1.0 mL/min.
Results:
Even though HPLC is more sensitive for diosgenin analysis, the developed RP-HPLC method should offer a fast, accurate, simple, and inexpensive alternative approach for the quantitative detection of diosgenin, gallic acid, and 18β-glycyrrhetic acid.
Conclusion:
The process described in this paper produces the same amount of pure diosgenin as other reported methods.