Polyisoprenoid composition in chengham (Scyphiphora hydrophyllacea, Gaertn. f., Rubiaceae)

Chengham (Scphyphora hydrophyllacea Gaertn. f., Rubiaceae) is a mangrove shrub or tree commonly in landward mangrove zones having some biological activities. All parts of S. hydrophyllacea have shown some components benefits to biological and pharmaceutical properties. This present study aimed to analyze the distribution and composition of polyisorenoid in senesnece leaves, fruits and branches of S. hydrophyllacea. Polyisoprenid patter was evaluated by two-dimensional thin layer chromatography (2D-TLC) approach. The total lipid (TL) content of five samples ranged from 5.7 mg/g in branches to 10.0 mg/g in the old leaves. Additionally, the percentage of polyprenol of senescence leaves slightly abundance compare to dolichol in the polyisoprenoid presence. Overall, polyprenol was found in higher content than dolichol in the yellow leaves. Results of this study found the type-II of S. hydrophyllacea fruits was in line with earlier reports on the plant fruits. The present study provided the occurrence of polyprenols and dolichols in S. hydrophyllacea tested organs.


Introduction
Ecology feature of chengham (Scphyphora hydrophyllacea) grows on mud, sand and coral substrates on the edge of mangrove land or on embankments [1].This minor mangrove seems to be intolerant of long-term inundation of fresh water and habitually occupies locations that are commonly inundated by tides [2]. It has been reported to grow in locations that are not suitable for colonization by other mangrove species. The inflorescences are present all year round, possibly self-pollinated or by insects [3]. Nectar is produced by glandular discs at the base of the flower crown [4]. A lot of fruit is produced, but seed reproduction is relatively low. The fruit is well adapted to dispersal by water because of its light, buoyant skin [4,5].
Scphyphora hydrophyllacea distributed from India, Sri Lanka, Malaysia, all of Indonesia, Papua New Guinea, Philippines, Solomon Islands and Tropical Australia [3]. Larger pieces of S. hydrophyllcea wood are used to make household items such as spoons, while smaller pieces are used to make fence posts and firewood [4]. The etnobotanical use has been reported as an extract of the leaves could be used for stomachaches [3][4][5]. Furthermore, anti-hepatocarcinogenic and anti-oxidant  [6]. S. hydrophyllacea has the potential to be developed as antimicrobial and anticancer compounds [7,8]. Despite the importance characterization of ecological and pharmacological in S. hydrophyllacea, no information on the polyisoprenoid composition of this plant, therefore this work purposed to analyze the distribution and composition of olyisorenoid in senesnece leaves, fruits and branches of S. hydrophyllacea

Plant materials
The leaves (young, old, and yellow), fruits and branches of Scyphyphora hydrophyllaceae Gaertn. F. (Rubiaceae), were sampled from Lubuk Kertang, North Sumatra, Indonesia. These minor mangrove are tree or shrub height up 3 m. These plants have simple leaves ( Fig. 1 A, B, C) and fruit diameter 0.4-0.5 cm, green to brown color, glabrous, and deeply groveed, longitudinally like small pinion-gear ( Fig. 1D) with maroon branches (Fig. 1E).

Extraction of polyisoprenoid
The previously described procedures for isolating polyisoprenoids was applied to this work [9,10]. The dried tissue was ground into a fine powder and absorbed in 30 ml of chloroform/methanol (2/1, v/v) solvent for two days, then it's over. The un-saponifiable lipids extracted from the tissue sample with hexane and the organic solvent were dried and re-dissolved in hexane.

Determination by two-dimensional thin layer chromatography (2D-TLC)
The longitudinal edge of the 1D-TLC plate was 1 cm wide, and the concentration zone of a reversedphase C-18 glass TLC was tightened using two magnetic bars (4.0 1.1 0.8 cm) that met each gel phase.. The bound TLC plate was then developed vertically to the 1D in order to transfer polyprenol or dolichol to the reversed-phase TLC plate's concentration zone. The 2D-TLC reversed-phase C-18 silica gel was done for approximately 40 minutes with acetone as the solvent. Using a CanonE-470 series printer, the developed chromatographic images were captured and scanned digitally.

Polysioprenoid profile and composition
The search for polyisoprenoids in the senescence leaves, fruits, and branches of S. hydrophyllacea in North Sumatra province, Indonesia, was carried out using 2D-TLC [11,12], which revealed a clear separation of polyprenols and dolichols based on carbon chain length.  Table 1 illustrates the total lipid (TL) content of five samples ranged from 5.7 mg/g in branches to 10.0 mg/g in the old leaves. Furthermore, the percentage of polyprenol in senescence leaves was slightly higher than that of dolichol in the presence of polyisoprenoid. (Table 1). This work was supported by foregoing findings on the type-II of aging leaveas of mangroves in Excoecaria agallocha, Lumnitzera racemosa, Rhizophora stylosa, R. lamarkii [13], K. obovata yellow leaves [14], Chromolaena odorata old leaves [15].

Analysis polyisoprenoid by 2D-TLC
The polyprenols and dolichols found in the leaves, fruits, and branches were discovered to belong to a precedingly established categorization, type-II. In the leaves, the composition became one group, group-II, as they showed the occurrance of both polyprenols and dolichols.
Furthermore, the finding of this study found the type-II of S. hydrophyllacea fruits was in line with earlier reports on the fruits of Elaeis guineensis [16], Nephellium lappaceum fruits [17], fruits of Amorphophallus paeoniifolius , Guettarda speciosa, and Jatropha curcas categorozed as type-II of polyisoprenoid [18], and Thespesia populnea fruits [19]. By contrast several plant fruits were reported as type-I such as, Barringtonia racemosa and Xylocarpus granatum [19].  Table 2. Carbon-chain lengths of polyprenol and dolichol of S. Hydrophyllacea.
Furthermore, the presence of ficaprenols (C60) in old leaves and fruits of S. hydrophyllacea samples was investigated. Furthermore, the dolichol content of the leaves was found to be unaffected by senescence (Figure 2). Longer polyprenols were found in yellow leaves, but no longer carbon-chain of dolichols were found. A similar report was detected in R. apiculata and R. lamarkii old leaves [13] and X. granatum fruits [19]. By contrast longer carbon-chain of polyprenols as well as dolichols have been reported in E. agallocha and L. racemosa yellow leaves [13]. The carbon chain length of polyprenols and dolichols due to the habitat and environmental differentiation and biosynthetic pathways to produce a variety of polyisoprenoids in tropical and subtropical plants [13][14][15]20].

Conclusions
The present study provided the presence of polyprenols and dolichols in S. hydrophyllacea detected tissues.