Development and Validation of HPLC Method for Quantification of Plumbagin in Plumbago Zeylanica L. Roots

Abstract

Background An RP-HPLC (Reverse Phase-High-performance liquid chromatography) method for the quantitative estimation and validation of the plumbagin in the methanolic fraction of Plumbago zeylanica L. was developed.

Method For achieving good separation, the RP-HPLC method was carried out with reverse phase C18 column, using methanol and water as mobile phase in the ratio of 65:35 (v/v), at the flow rate of 1 mL/min. The detection wavelength was set at 265 nm.

Results The retention time of plumbagin was found at 7.5±0.2 min. The coefficient of determination of plumbagin was found to be (r²) 0.9985 and equation Y=23148x+4327. The LOD and LOQ were found to be 34.06 and 113 ng/mL, respectively.

Conclusion The developed method was accurate, specific, precise, and reproducible. This RP-HPLC may be useful for quantitative estimation of the chemical constituents present in the plant extract as well as the quality assessment of the herbal product.

Key words

Plumbagin - Plumbago zeylanica L - High-performance liquid chromatography - method validation - quality control.

Publication History

Article published online:
23 February 2023

• References

• 1 Arpita R, Navneeta B. A review on pharmaceutically important medical plant: Plumbago zeylanica . J Ayu Herb Med 2017; 3: 225-228
• 2 Rajakrishnan R, Lekshmi R, Benil PB. Phytochemical evaluation of roots of Plumbago zeylanica L. and assessment of its potential as a nephroprotective agent. Saudi J Biol Sci 2017; 24: 760-766
• 3 Chauhan M. A review on morphology, phytochemistry and pharmacological activities of medicinal herb Plumbago zeylanica Linn. Int J Pharmacog Phytochem Res 2014; 3: 95-118
• 4 Shukla B, Saxena S, Usmani S. et al. Phytochemistry and pharmacological studies of Plumbago zeylanica L.: a medicinal plant review. Clinical Phytosci 2021; 7
• 5 Aleem M. Anti-inflammatory and antimicrobial potential of Plumbago zeylanica L: a review. J Drug Deliv Ther 2020; 10: 229-235
• 6 Gabriel O, Ademuyiwa O, Lasisi AA. et al. Free radical scavenging activities of extracts and bioactive constituents from the roots of Plumbago zeylanica (Linn.). Eur J Biol Med Sci Res 2019; 7: 21-33
• 7 Bloch K, Parihar VS, Kellomäki M. et al. Natural Compounds from Plumbago zeylanica as Complementary and Alternative Medicine. In: Chakraborti S. (eds) Handbook of Oxidative Stress in Cancer: Therapeutic Aspects. Springer; Singapore: 2022
• 8 Koetz M, Santos TG, Rayane M. et al. Quantification of atropine in leaves of Atropa belladonna: development and validation of method by high-performance liquid chromatography (HPLC). Drug Anal Res 2017; 1: 44-49
• 9 Nag M, Kar A, Chanda J. et al. RP-HPLC analysis of methanol extract of Viscum articulatum. J Ayurveda Integr Med 2020; 11: 277-280
• 10 Govindarajan R, Tejas V, Pushpangadan P. High-performance liquid chromatography (HPLC) as a tool for standardization of complex herbal drugs. J AOAC Int 2019; 102: 986-992
• 11 Engsuwan J, Waranuch N, Limpeanchob N. et al. HPLC methods for quality control of Moringa oleifera extract using isothiocyanates and astragalin as bioactive markers. Science Asia 2017; 43: 169-174
• 12 ICH-Q2A, Text on validation of analytical procedures. In: Harmonized tripartite guideline prepared within the international conference on harmonization of technical requirements for the registration of pharmaceuticals for human use. Geneva: 1994
• 13 ICH-Q2B (1996) Validation of analytical procedures: methodology. In: Harmonized tripartite guideline prepared within the international conference on harmonization of technical requirements for the registration of pharmaceuticals for human use. Geneva: 1996