Piper chaba (locally known as “Choi Jhal”) is used traditionally as spices and folk medicine in different parts of Bangladesh. One of the most important bioactive compounds in this plant is piperine. In this study, the amount of piperine in P. chaba root and stem was investigated and the optimal solvent for piperine extraction at room temperature was also studied. High performance liquid chromatography (HPLC) was operated using a reverse phase column where methanol and water (70:30) were used as mobile phase. The detection was performed using photo diode array (PID) detector at a wavelength of 345 nm. The standard piperine showed linearity between 0.005 % and 0.04 % and the correlation co-efficient found for the linearity was 0.9933. The percentage of relative standard deviation (RSD) for both retention time and peak area were less than 2.0 %. The theoretical plate number (N) > 3000 and a tailing factor (T) < 1.5 were found in the acceptable range. The recovery percentage (%) of standard piperine was 99.16 %. Low value of co-efficient of variation and standard deviation are recognized for high precision of the method. The highest amount of piperine was found in root extracted with methanol (MR) amounting to 1.75 % in the root powder. The maximum amount of piperine in the stem was 1.59 % when extracted with methanol (MS). The piperine quantification in other extract like n-hexane root (HR), ethyl acetate root (ER), n-hexane stem (HS), ethyl acetate stem (ES) were 0.76 %, 1.69 %, 0.33 % and 1.46 % respectively. Methanol has given the highest yield of piperine compared to ethyl acetate and n-hexane for both root and stem. The developed method was simple, rapid, economic and validated in terms precision, accuracy and recovery. This selective method is found to be repeatable, accurate and successfully utilized for the Piper extract in marketed and pharmaceutical samples with well chromatographic conditions. The ethyl acetate extract of root and stem showed promising DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical scavenging activity with an IC50 value of 39.62 ± 0.95 μg/mL and 43.85 ± 1.50 μg/mL respectively. The study reports potential antibacterial activity and antifungal activity of P. chaba root and stem extracts. These outcomes revealed that different extracts of P. chaba may be used as natural preservatives.
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