Authors:Aziz Ahmed, Sayeed Ahmad, Mahfooz Ur-Rahman, Tamboli E. Tajuddin, Ranbir Verma, Mohd Afzal and Prahlad Singh Mehra
Bacoside A, a triterpenoid saponin, is a major constituent isolated from Bacopa monnieri (L.) Wettst. (Scrophulariaceae), used as a memory enhancer. Bacoside A and B are active ingredients in Bacopa herb and have antioxidant and hepatoprotective activities
A new rapid, simple, and economical high-performance thin-layer chromatographic (HPTLC) method was developed and validated for densitometric quantitative analysis of bacoside A in powdered leaves from different geographical regions of India.
Materials and methods
An amount of 10 mg mL−1 methanol extract of powdered leaves from different geographic regions was used for sample application on precoated silica gel 60 F254 aluminum sheets. Standard bacoside A (1 mg mL−1) was used for calibration curve. HPTLC separation was performed on percolated silica gel aluminum plate 60 F254 (20 cm × 10 cm with 0.2 mm thickness) as a stationary phase using ethyl acetate–methanol–water (4:1:1) as the mobile phase. Quantification was achieved by densitometric analysis at 598 nm over the concentration range of 500–4000 ng band−1.
Compact and well-resolved bands for bacoside A from powdered leaves of different geographic regions were found at retardation factor (Rf) 0.53 ± 0.02. The linear regression analysis data for calibration curve showed good linear relationship with regression coefficient r2 = 0.9996 and r2 = 0.99810 with respect to peak area and peak height. The method was validated for precision, recovery, and robustness as per the International Conference on Harmonization (ICH) guidelines. Variation in quantitative analysis of bacoside A in powdered leaves sample from different geographic regions was found by HPTLC method.
Discussion and conclusion
The highest and lowest content of bacoside A in powdered leaves sample from Jammu and Kerala regions, respectively. The variety of B. monnieri in Jammu is superior to other regions of India. The proposed developed HPTLC method can be applied for the quantitative determination of bacoside A in powdered leaves of plant and its formulation.
Bacopa monnieri (L.) Pennell, commonly known as Brahmi, is an important medicinal plant of the Ayurvedic System of Medicine and is categorized as Medhya Rasayana, used for boosting memory. Bacoside A and B (saponins) are major bioactive compounds of the plant responsible for memory enhancement. Consistent quality and quantity of the bioactive compounds are a must to produce highly efficacious herbal drugs. The phytochemical diversity due to ecogeographical variations affects the persistent therapeutic effect. In the present study, quantification of the major bioactive principle of B. monnieri, bacoside A, was performed in order to evaluate chemodiversity in the plants collected from different ecogeographical locations. The analysis of 75 accessions showed a wide range of significant variation in bacoside A content. Further, genetic diversity study was carried out between high- and low-yielding accessions to correlate the effect of the genetic composition on chemical diversity. The cluster analyses of amplified fragment length polymorphism (AFLP) data of high and low bacoside A producing plants using dendogram and principal coordinate analysis (PCoA) revealed no specific clustering. The genetic diversity parameters calculated also support less genetic differentiation between low- and high-yielding accessions. The results clearly indicate that there is a prevalence of agroclimatic conditions on differential bacoside A accumulation in B. monnieri. The study will help to produce quality plant material to be used as phytomedicine.
Authors:Qianqian Xing, Qing Fu, Yu Jin and Xinmiao Liang
widely used application was the analysis and purification of diastereomeric saponins [ 29 ]. Agrawal et al. developed the SFC method for standardization of isolation of bacosideA 3 and bacoside II in Bacopa monnieri extract [ 30 ]. However, studies on