RP-HPLC Characterization of Lupenone and β-Sitosterol in Rhizoma Musae and Evaluation of the Anti-Diabetic Activity of Lupenone in Diabetic Sprague-Dawley Rats

Abstract

With the aim of characterizing the active ingredients lupenone and β-sitosterol in Rhizoma Musae samples a reversed-phase HPLC method for the separation of these two compounds in Rhizoma Musae samples was developed (regression coefficient > 0.9996). The method was further applied to quantify lupenone and β-sitosterol content in Rhizoma Musae samples cultured in different growth environments. Different variables such as geographical location, growth stage, and harvest time, demonstrated differential effects on lupenone and β-sitosterol levels. Moreover, we determined the optimum conditions for cultivation and harvesting of Rhizoma Musae herbs. Lupenone administration caused a significant reduction in fasting blood glucose (FBG) levels in diabetic rats at doses of 1.78, 5.33, and 16.00 mg·kg for 14 days, the glycated hemoglobin (HbA1c) levels of diabetic rats also significantly reduced at doses of 5.33, and 16.00 mg·kg, indicating a robust antidiabetic activity. To our knowledge, this is the first report of an optimized HPLC method successfully applied to quantify lupenone and β-sitosterol, and its applicability in optimizing Rhizoma Musae growth. Animal experiments also showed for the first time that lupenone from Rhizoma Musae has anti-diabetic activity.

Antihyperglycemic Activity of Banana (Musa nana Lour.) Peel and Its Active Ingredients in Alloxan-Induced Diabetic Mice

“…According to reports, lupenone can inhibit α -glucosidase ( α -Glu) and protein tyrosine phosphatase 1B (PTP1B) activity in vitro [31,32], the two enzymes are associated with anti-diabetic, and recently study has found that lupenone has the anti-diabetic activity in diabetic sprague-dawley rats [33]. In this study, the results clearly showed that lupenone had a significantly antihyperglycemic activity in alloxan-induced diabetic mice with no animal deaths during the experiment.…”

Determination of Lupenone and β-Sitosterol in Rhizoma Musae by UPLC with DAD and ELSD

“…Comparing with the established HPLC method (19), the present method had a saved (85%) mobile phase, shorter (50%) analysis time and greatly improved resolution and sensitivity. This method was applicable to determination of a large number of Rhizoma Musae samples, and it can reduce pollution and protect the environment.…”

“…In this study, lupenone and β-sitosterol were selected as index components in Rhizoma Musae, because some literature has shown that the lupenone and β-sitosterol have anti-diabetic activity (18,19), so the two components were selected as index components in Rhizoma Musae to control the herb quality. For the simultaneous analysis of lupenone and β-sitosterol in Rhizoma Musae, a fast, reliable and sensitivity UPLC with DAD and ELSD method was developed, the method was validated in specificity, linearity, precision Y is the peak area in DAD chromatograms monitored at 206 nm, X is the compound concentration (µg/mL) injected, and y and x are the logarithmic values of area and concentration injected in ELSD chromatograms.…”

“…Lupenone and β-sitosterol are responsible for the anti-diabetic activity of Rhizoma Musae, and the high-performance liquid chromatography (HPLC) method for detecting lupenone and β-sitosterol has been established (19), but the analysis time is too long and sensitivity of the analysis is not suitable for simultaneous determination of the lupenone and β-sitosterol at low levels. The ultra-performance liquid chromatography (UPLC) is a new liquid chromatography (20); it can save mobile phase and improve resolution, speed and sensitivity of analysis.…”

“…The β-sitosterol has anti-cancer activity (15)(16)(17). Besides, lupenone and β-sitosterol show a good anti-diabetic activity in vivo (18,19).…”