Thin Layer Chromatography Fingerprinting and Clustering of Orthosiphon stamineus Benth. from Different Origins

Introduction: Orthosiphon stamineus has been widely used across Asian countries for the treatment of various diseases. The quality of herbal medicine determine its safety as well as efficacy; and geographical origin is important factor contributing on the quality of herb and its products. Materials and Methods: Thin Layer Chromatography (TLC) method combined with chemometric, Principal Component Analysis (PCA), has been employed to evaluate the quality of Orthosiphon stamineus leaves collected from eleven origins in Indonesia. Results: The results showed that mobile phase suitable for Orthosiphon stamineus was chloroform, dichloromethane, ethyl acetate (7:4:1). The method used has met the requirements of TLC system stability and precision. TLC-fingerprints analyzed with chemometrics showed an ability to discriminate Orthosiphon stamineus from various origins. PCA score plot of the first two principal components (PC) clearly distinguished 3 clusters of samples, whereas the loading plot of the first two PC showed that compounds with the Rf values of 0.0-0.1, 0.1-0.2, 0.2-0.3, and 0.9-1.0 are the most important compounds for clustering of samples. Conclusions: TLCfingerprint combined with the PCA was able to discriminate among the leaves of Orthosiphon stamineus originated from various locations. TLC-fingerprints analyzed with chemometrics can be used as an alternative of marker-oriented method to evaluate the quality of Orthosiphon stamineus.