Isolation and identification of Bengle (Zingiber cassumunar roxb) as a stimulant in phagocytic activity of macrophages

Immunomodulators are pharmacological agents that modify or regulate the immune system through stimulating the functioning of the immune system and, at the same time, inhibiting excessive immune responses. This study was conducted to determine the active compound in Z. cassumunar that is responsible for increasing the immune system based on the parameters of phagocytic activity. The isolation method began with fractionation, which involved extraction with ethanol and successive fractionation with hexane and chloroform. Z. cassumunar extract, hexane fraction, and chloroform fraction were tested on mice macrophage cells for their phagocytic functions. The phagocytic activity of macrophages was measured by active phagocytic cells (averagely 39.194 ±1.597, 27.923 ±2.941, and 62.090 ±6.947) and phagocytic index (in a row, averagely 47.513 ±2.844, 41.129 ±7.195, and 101.527 ±10.555). The results showed that the Z. cassumunar extract,  hexane fraction, and chloroform fraction exhibited more significant phagocytic activities of macrophages (p <0.05) compared with the normal group. Since the chloroform fraction showed the best result, this fraction was further separated by column chromatography. This procedure yielded five sub-fractions, namely F1, F2, F2C, F3, and F4. Based on the phagocytic activity testing, the results were as follows: (1) the active phagocytic cells of F1, F2, F2C, F3 and F4 were 18.860 ±3.191, 27.077 ±4.482, 15.749 ±3.026, 64.333 ±1.780, and 44.943 ±2.944, respectively, and (2) the phagocytic indices were 30.0249 ±3.4231, 44.5969 ±8.3646, 24.5597 ±5.4487, 102.7447 ±1.0806, and 76.5007 ±4.7293. Because F3 produced the best result, this subfraction was then identified using 1H-NMR and 13C-NMR. The identification results showed that F3 was (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol as an active compound