RP-HPLC analysis of methanol extract of Viscum articulatum

Viscum articulatum Burm. (Family: Loranthaceae) is commonly known as mistletoe. In ayurveda, the plant parts are used in "Kapha", "Vata", diseases of the blood, ulcer, and epilepsy. The plant parts are also used in urinary tract infection and wound infection. The plant contains five triterpenoids such as α-amyrin, lupeol, betulin, betulinic acid and oleanolic acid, exhibiting several pharmacological activities including antimicrobial, anti-HIV, antitumor, antiviral activity.

To ensure the content of uniformity of oleanolic acid, a RP-HPLC method has been developed for estimation of oleanolic acid in V. articulatum aerial part.

The RP-HPLC method was carried out in reverse phase C₁₈ column, using methanol and water as mobile phase in the ratio of 95:5 (v/v), at the flow rate of 1 mL/min. The pH of aqueous phase was adjusted 3.2 with 1% (v/v) glacial acetic acid. The λ_max was set at 210 nm.

The retention time of oleanolic acid was found at 21.5 ± 0.05 min. The linearity of the response was found to be 10-800 μg/mL. The coefficient of determinants of oleanolic acid was found to be (r²) 0.995 and equation Y = 19462X + 16,172. The LOD and LOQ were found to be for oleanolic acid (1.96% w/w) 0.197 ± 0.63 and 0.623 ± 0.87 μg/mL, respectively. 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.