A high-performance thin-layer chromatographic (HPTLC) method has been established for quantitative analysis of andrographolide, the major bioactive constituent of the immunomodulatory plant
Nees. The method is extremely sensitive, and is economical in consumption of sample (10 mg), solvent (40–50 mL methanol), and time (2–2.5 h). The method was validated for specificity, precision, and accuracy in accordance with International Conference on Harmonization (ICH) guidelines on technical requirements for registration of pharmaceuticals for human use. The relative standard deviation of intra-day and inter-day analysis was in the range 0.56–1.33% and accuracy, as determined by measurement of recovery, was, on average, 98%. In addition, the effect of drug solvent ratio on the quantification procedure was also studied in detail. In comparison with other methods for analysis of andrographolide, this method has the advantage of minimizing consumption of expensive drug samples, organic solvents, and other resources.
Authors:Astha Varma, Harish Padh, and Neeta Shrivastava
Quantitative evaluation of phytochemical diversity in Adhatoda vasica Nees populations from five different ecogeographical regions was performed using a highly sensitive, robust and economic high performance thin layer chromatography (HPTLC) procedure. The method was validated for specificity, precision and accuracy as per the guidelines affirmed by International Conference on Harmonization of technical requirements for registration of pharmaceuticals for human use (ICH). The findings support the existence of distinct natural chemotypes within the species. Two out of the five regions had about three times higher mean vasicine content than the remaining three regions. The comparative phytochemical fingerprint profiles were quite similar except for the concentration variation seen for different alkaloids. As the quinazoline alkaloids are the biologically active compounds in A. vasica, the samples that accumulate high levels of alkaloids seem to be promising for further propagation.
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.