Abstract
Cassia siamea leaf has been proven in vitro and in vivo to have a strong antimalarial activity with Cassiarin A as its active compound. To obtain a source of C. siamea medicinal plant with high level of active antimalarial compound (Cassiarin A), a valid method for determining Cassiarin A level is needed. For this reason, this research conducts the validation of the Cassiarin A content with determination method using thin-layer chromatography (TLC) densitometry which includes the determination of selectivity (Rs), linearity (r), accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ). Cassiarin A was chromatographed on silica gel 60 F254 TLC plate using chloroform : ethanol (85 : 15 v/v) as a mobile phase. Cassiarin A was quantified by densitometric analysis at 368 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r = 0.9995. The method was validated for precision, recovery, repeatability. The minimum detectable amount was found to be 0.0027 μg/spot, whereas the limit of quantitation was found to be 0.008 μg/spot. The results of this validation are then used to determine the Cassiarin A level of C. siamea leaf from various regions in Indonesia. Based on the results of the study, it can be concluded that the TLC-densitometry method can be used to determine level of the Cassiarin A compound with the advantages of being fast, easy, accurate, and inexpensive. In addition, it showed that C. siamea leaves from Pacitan have the highest level of Cassiarin A compared to other areas studied.
Highlights
Cassia siamea L. is one of the plants in Indonesia that has traditionally been recognized for its ability to treat malaria. is plant grows better in the lowlands, with low to high rainfall, an average temperature of 20°C–31°C, and with a four to eight months dry season, and spreads to Java at an altitude of less than 1000 masl [1]
Other in vitro and in vivo antimalarial studies on C. siamea leaf which in Indonesia is known by the name Johar successfully obtain new active antimalarial compounds namely Cassiarin A and Cassiarin B with the highest antimalarial activity possessed by Cassiarin A compound [5]
Cassiarin A as an antimalarial bioactive compound from the C. siamea leaf was used as a marker compound in this study
Summary
Cassia siamea L. is one of the plants in Indonesia that has traditionally been recognized for its ability to treat malaria. is plant grows better in the lowlands, with low to high rainfall (optimum around 1000 mm), an average temperature of 20°C–31°C, and with a four to eight months dry season, and spreads to Java at an altitude of less than 1000 masl [1]. Other in vitro and in vivo antimalarial studies on C. siamea leaf which in Indonesia is known by the name Johar successfully obtain new active antimalarial compounds namely Cassiarin A and Cassiarin B with the highest antimalarial activity possessed by Cassiarin A compound (see Figure 1) [5]. Several studies have reported that locations or regions with different latitudes, longitudes, average temperatures, and climatic conditions affect the production of secondary plant metabolites and have an effect on their biological activity [9,10,11,12,13]. To get a source of medicinal plants with high levels of active antimalarial compounds (Cassiarin A), a valid method for determining levels of Cassiarin A is needed. Quantification of Cassiarin A as an antimalarial active compound in C. siamea leaf using thin-layer chromatography (TLC) densitometry to date has never been reported
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