Dendrophthoe Petandra L. Miq.s Progesterone-Like Effect

Dendrophthoe Petandra L. Miq.s Progesterone-Like EffectABSTRACTThe in-vivo convey on healthy adult female rat were known that suck out methanol of Dendrophthoe Petandra L. Miq by intramuscular injection (w/v) at dose 100 mg/kg body weight for four age (s.d.d.) can be produce progesterone hormone two seasons more than usual levels. That phenomenon seemingly indicates that plant as an Indonesia namely Benalu duku were probably consist of steroid with carbon copy bonding meanss at bend 1st to 21st namely pregnane derivate. These research aims to determined specialisedity active cognitive contents of methanol crude option Benalu duku leaves with pharmacodynamics issuing like progesterone hormone. Pulverized Benalu duku leaves at 400g was draw ined with 2 L methanol pro epitome circle by shaker maceration method at 200C during the 72 hours. Semi solid crude plagiarize were drying by vacuum dried apparatus and UV exposure at 15 minutes and analysis by FT-IR and LC ESI MS. R esearch result obtained that active substances with primary(prenominal) chemical organise of pregnane derivate were progesterone at somewhat 30%, medroxy progesterone ethanoate at about 66%, megestrol acetate rayon at about 3% and 1% dydrogesterone.Key words Benalu duku, pregnane derivate, anabolic steroid, leaves perpetrate FT-IR, Methanolic leaves extract LC-ESI MS.IntroductionDendrophtoe Petandra L Miq (Benalu duku) as a parasites plant growing on Lancium demesticum were known have a several essential active substances and exertionual for treated proliferation cell (NURAINI et al., 2000, ROOSTANTIA et al., 2000 and RATNA et al., 2007). Some researcher reported that plants have a beneficially active substance like alkaloid, flavonoid, polyphenol, terpenoid and free steroid (ARIFA et al., 2009). New research report explained that crude methanolic extract of Benalu duku leaves can be increasing progesterone hormone two times more than usual levels, but giving follicle-stimul ating hormone levels on female adult rat (BAMBANG, LAZUARDI, 2014). The last research report indicate that plants have a several substances with characterize as a progesterone like way out.Characterization progesterone like effect was known have a special chemical main structure namely pregnane derivate with consist of carbon at 1st to 21st at their double bonded chain structure or (8S,9S,10S,13R,14S,17S)-17-ethyl- 10,13-dimethyl-2,3,4,5,6,7,8,9,11,12, 14,15,16,17-tetradecahydro-1H- cyclopenta a phenanthrene. Identically of their main structure of pregnane derivate were known as an anabolic hormone and usefully for treatment prostate cancer in adult man (RANDOX LABORATORY LTD, 2013). Pregnane derivate were known have some specific anabolic steroid structure i.e cortisone, hydrocortisone, progesterone, medroxy progesterone acetate, megestrol acetate, 17- - hydroxyl progesterone acetate, dydrogesterone.Other herbal medicine with anabolic steroid action was raise at Mexico namely Co dices plant. That plant was similar with Benalu duku as a parasites plant growing at subtropics seasons (LERNER, ARNWINE, 2002). With analog to Codices plant in Mexico, might be benalu duku leaves after extracted by methanol were containing specific anabolic steroid. By background research at above, will be determined progesterone like effect substance like progesterone, medroxy progesterone acetate, megestrol acetate, dydrogesterone from base on overview of identic scintilla ion of LC-ESI MS and function compound of FT-IR.Materials and methodsBenalu duku leaves were obtained from Muara Enim neighbourhood of west Sumatera-Indonesia and identification as a Dendrophthoe Petandra L. Miq at Botany Research Institute at Tangerang capital of Indonesia Indonesia as described by BAMBANG, LAZUARDI (2014). Reference substance of progesterone was obtained from Sigma-aldrich product 46665 batch SEBA XV. Reference substance of medroxy progesterone acetate was obtained from Harsen pharmaceutic al Industry Jakarta Indonesia at pharmaceutics grade. Megestrol acetate reference corporeal was obtained from Sigma Aldrich at product 46420 batch SZB9173XV. Dydrogesterone reference material was used from European Directorate for quality medicines and health care, France by catalog code Y0001004.A 400 gram of pulverize benalu duku leaves were extracted on 2 L of methanol pro analysis on to the 1 L separated flask during the 72 hours (movement shaker maceration method at 200C). The semi-solid of crude extract were dried on vacum drying and exposure to uv hoy (15 minutes) for kept contaminate from the mold and some bacteria. Some amount of semi-solid crude extract benalu duku leaves was determined specific molecule structure compound by FT IR PerkinElmer Frontier 89485 and LC-ESI MS Accela TSQ Quantum Acess Thermo. The semi-solid crude extract methanol of benalu duku leaves was determined to molecule function of progesterone like effect substances at 400 cm-1 to 4000 cm-1 . exploi t control of LC-ESI MS were adjusted as follows column hypersil Gold 0.2 m length 10 cm gradient method with mobile phase at bottleful A 0.1% in Aqua bidest and bottle B 0,1% formic acid in acetonitrile pro HPLC. The gradient was perform at 35% to70% of B in 20 minute at 254 nm UV detector. Automatic sucking and injection might was adjusted at 10 l, flushing capacity 400 l with amphetamine at 100l.second-1, velocity injector 8 l.second-1, height of suction apparatus at about 2 ml capacity autosample phial at 1.2 ml. Temperature column was adjusted at 220C at maximum pressure pump 1250 PSI and stabile in 10 BAR (THERMO ELECTRON CORPORATION, 2007).ResultsAnalysis Spectrum infra-red of crude extract methanol Benalu duku leaves obtained that stretching redolent(p) compound specific overtone was presented at 1650 to 2000 cm-1. Especial in 1550 cm-1 to 1600 cm-1 was appeared stretching aromatic compound (1,s). In 1450 cm-1 to 1500 cm-1 was appeared s orbital of stretching aromatic co mpounds. At finger print area wavelength number of 600 cm-1 to 900 cm-1 and yard cm-1 to 1300 cm-1 were indicate of flexible carbon atom and hydrogen outside of area (k orbital) and inside of area (l orbital). To analyze similarities of infra-red spectrum between reference material vs., crude extract methanol of Benalu duku leaves at high intensity (%T) indicated that wavelength number of 1400 cm-1 to 1050 cm-1 and 1600 cm-1 to 1660 cm-1 were identic both of them. Even at low intensity (%T) on the finger print area 750 cm-1 to gravitational constant cm-1 were identic infra-red spectrum between reference materials vs., crude extract methanol benalu duku leaves. display panel 1 at bellow was presented specific function molecules of crude extract methanol benalu duku leaves after assessment by FT-IR by references of NOERDIN (1985).Analysis LC ESI-MS by Triple Stage Quadruple Mass Spectrometers were obtained that compound of progesterone like effect in extract methanol Benalu duku leaves were identic to reference materials of medroxy progesterone acetate, progesterone, megestrol acetate and dydrogesterone. Table 2 at bellow explained that computer storage time and ion molecule of ESI (m/z) as dependent variable of extract methanol Benalu duku leaves was similar to store time and ion molecules of ESI references material.Table 1. Infra-red spectrum (cm-1) and intensity (%T) crude extract benalu duku and reference materials of progesterone like effect substancesExtract benalu dukuProgesteroneMed. Prog. acetMegestrol acetateDydrogesteroneIdentified function group,cm-1 %Tcm-1%Tcm-1%Tcm-1%Tcm-1%T3500-340015-113436.69723434,340,113436,3910,163435,72-3374,3414,17-14,580-H2927,3312,872969,05-292510,25-8,22 abstracted2946,49-2927,6612,04-12,172989,16-2930,195,05-13,35 asymmetrical vibration, stretching C-H1632,8817,151699,13-1616,1749-8,831638,951,051664,2-1629,096,49-10,91659,51-1620,821,24-2,94Stretching aromatic ring (1,s)1458,0718,491438,7615,451401,193,291458,71 4,881452,0614,48Stretching aromatic ring (s)1272,2219,211279,25-1268,8917,85-18,67Absent1269,8310,11277,712,4Flexible C-H on orbital (l)1203,0919,361204,9114,97Absent1206,3114,181193,17,97Alkene, R-OH, stretching vibration C-H, aromatic1169,4419,731179,06-1162,4817,64-16,36Absent1166,4116,271174,79-1162,5817,08-14,51-CHCH-(trans),R-CH21062,8119,171116,3219,961112,603,281083,18-1058,6216,29-15,811064,1621,79CCH2, mono substitute alkenes826,5523,84871,1817,15Absent877,33-796,7814,9-21,26831,21-793,8624,45-23,51Tri substitute alkenes, meta-di subtitute Benzene (aromatic)721,4823,67778,14-687,3121,77-21,17Absent755,83-713,1920,62727,7723,21cis-di substitute alkenes, mono substitute benzene, meta-disub. benzene (Aromatic)639,2123,8687,3121,17644,792,57634,9420,65630,9918,55cis-di substituted alkenes (vinyl, C-H)Table. 2. Result analysis retention time and molecules ion of ESI between crude extract methanol benalu duku leaves and reference material as medroxy progesterone acetate, progest erone, megestrol acetate and dydrogesteroneAnalytesWeight (g/ml)AreaRetention time(minutes)Electron Spray Ionization (ESI)(m/z)Crude extract benalu duku leaves0,06462270444.40387.000Medroxy progesterone acetate0,15995854.40387.000Crude extract benalu duku leavesFigure 1 to Figure 4 at below obtained that LC ESI-MS of extract methanol benalu duku leaves were containing identic active substances of medroxy progesterone acetate, progesterone, megestrol acetate and dydrogesterone.DiscussionAnalysis by retention time and ESI (m/z) of LC ESI-MS were apparently just two of progesterone like effect substances on crude extract methanol benalu duku leaves that rattling identic substances to reference material namely medroxy progesterone acetate and progesterone. Other progesterone like effect substances as a megestrol acetate and dydrogesterone on crude extract methanol benalu duku leaves were mostly identic to reference material of megestrol acetate and dydrogesterone by different shift at 0.01 and 0.6 retention time of reference materials. Difference in retention time shift is closely linked nature of polarity samples bound to the column Hypersil Gold of LC. so the most likely molecular structure of megestrol acetate and dydrogesterone on methanol extract of benalu duku no likeness to 100% identically as reference materials of megestrol acetate and dydrogesterone. Analysis to compare between the values of molecule ion fragments (m/z) and ESI (m/z) reference materials of fourth progesterone like effect substances versus identically substances of progesterone like effect on crude extract methanol benalu duku leaves were apparently have a differences values (p