The oml administration of the chloroform extract of Parkia speciosa to alloxan-induced diabetic rats produced a

significant (p<0.01) decrease in blood glucose levels. The hypoglycemic response was approximately proportional to

the square root of the dose given. The hypoglycemic activity of the extract reached a maximum 2-5 hours rifter oral

administration of the extract and lasted for at least 24 hours

Jamaludin, Fathaiya

Mohamed, Suhaila

Universiti Putra Malaysia Institutional Repository

PertanikaJ Trap. Agric. Sci. 16(3): 161-165(1993) ISSN:OI26-6128© Un i\'ersi ti Pertan ian ~lalaysia PressHypoglycemic Effect of Extracts of Petai Papan(Parkia speciosa, Hassk)FATHAIYAJAMAL DIN and SUHAILA MOHAMED'Department ofrood Science,p'aculty ofFood Science and Biotechnology,Universiti Pertanian lVIalaysia, 43400 Serdang, Selangor, MalaysiaABSTRAKPentadbimn extrak klorofonn jJetai melalui mulut, dapat menumnkan dengan ketam (p<O. 01) kandongan glukosdalam darah tikl.ls yang di kencing manis oleh alloxan. Tindakan hypoglysaemik ini berkadaran dengan puncakllasa dl/a dos yang diberi. Tindalwn hypoglysaemik adalah mixima selepas 2-5 jam pengambilan ekstrak tersebutmdallli ml.llut dan I~elwl selama sekumng-kumngnya 24 jam.ABSTRACTThe oml administration of the chloroform extract ofParkia speciosa to alloxan-induced diabetic rats produced asignificant (p<0.01) decrease in blood glucose levels. The hypoglycemic response was approximately proportional tothe square root of the dose given. The hypog(ycemic activity of the extmct reached a maximum 2-5 hours rifter oraladministration of the extract and lasted for at least 24 hours.Keywords: Parkia speciosa, antidiabetic, hypoglycemic, oral administration, rats, chloroform extract,dose-responseINTRODUCTIONPetai (Parkia speciosa) is a Southeast Asianlegume of the Mimosae subfamily, whose seedsare consumed as a condiment or vegetable,,,,ith rice, for its unique Shiitake mushroom-like flavour. When taken in excess it gives astrong onion-like smell, which is excreted bythe body in the urine, the sweat and the faeces.Sometimes petai is eaten because it is believedto have anti-diabetic and anti-hypertensiveactivity.Petai has been used in traditional medi-cine for its antibacterial effects on kidney,ureter and urinary bladder. The antibacterialand antifungal compounds were found to becyclic polysulfides, whose structures wereestablished as 1,2,4-trithiolane, 1,2,4,6-tetrathiepane, 1,2,3,5,6-pen tathiepane (lenthio-nine), 1,2,4,5,7,8-hexathionane and a pen-ITo whom all correspondence should be addressed.tathiocane (Gmelin et al. 1981). Dichros-tachin ic acid, djenkolic acid and thiozolidine-4-carboxylic acid were qlso iden tified(Holzman et al. 1982). Thiozolidine-4-car-boxylic acid has been successfully used experi-mentally and clinically as an anti-cancer agent(Pandeya 1972). Djenkolic acid has beenknown to cause blockage of the urinarytubules due to its low solubility, resulting inpain, haematuria and even death. P. speciosaseeds also contain significant minerals, vita-mins, protein and fat, while having a lowerantinutrient content compared to soya bean(Suhaila et al. 1987).This research was undertaken to investi-gate the hypoglycemic effect of P. speciosa onnormal and alloxan-induced diabetic rats,because petai is eaten by diabetics for thatpurpose.FATHAIYAJAMALUDIN AND SUHAILA MOHAMEDMATERIALS AND METHODSPreparation ofExtractsTen kg of fresh petai pods were obtained fromthe local market. The seeds were separated fromthe pods. Both portions were air dried, groundto a powder and extracted sequentially andexhaustively with petroleum ether, diethyl ether,chloroform, dichloromethane, ammoniacalchloroform and methanol. The solvents werecompletely evaporated off with a rotary evapora-tor to obtain the extracts.Experimental ProcedureHealthy Sprague Drawley rats of mixed sexes(weighing 200-450 g) were intravenously injectedwith 60 mg/kg alloxan (2,4,5,6-Tetra oxy pyrimi-dine) to induce diabetes within 40-48 hours(Lundquist and Rerupa 1967). The dry extractsof petai were orally fed to 24-hr-fasted normaland alloxan-induced diabetic rats at a dose levelin the range of 25-500 mg extract/kg BW (bodyweight), together with 1 g glucose/kg BW of rat.Coadministration of glucose with the extract wasdone to cause hyperglycemia. Both diabetic andnormal rats treated orally with 5 ml saline and 1 gglucose/kg BW were observed for comparison.Blood samples were taken hourly for the first 11hours and again 24 hours after the administra-tion of the extracts. Blood was obtained from thetail vein by using heparinised microhematocritcapillary tubes (Riley 1960).A nalysis ofBlood GlucoseThe plasma glucose level was determined by glu-cose oxidase method (Roche Glucose test kit 007 1011 3) where D-glucose is specifically oxi-dised to gluconic acid and hydrogen peroxide byglucose oxidase. The generated hydrogen per-oxide converts O-dianisidine, by the catalyticaction of peroxidase to the red-brown semi-quinone. The colour intensity is directly propor-tional to the glucose concentration and is mea-sured spectrophotometrically. 0.02 ml of serumwas used for glucose assay and compared with0.02 ml standard D glucose solution.Statistical AnalysisThe data were statistically analysed using analysisof variance (ANOVA) , Duncan's multiple rangetest (DMRT) and regression analysis on MSTATcomputer program.RESULTS AND DISCUSSIONResults showed that only the chloroform extracts(l g/kg body weight) from both the empty podsand seeds of petai had a strong hypoglycemic activ-ity on diabetic rats (Fig. 1). Blood glucose level attime zero is the blood glucose level just after theoral administration of extracts/saline and glucose.ANOVA analysis showed significant differencesbetween chloroform extracts of both the seeds andpods (p<O.Ol), and extracts from other solvents··1 !- J.. ."I• I··········~·f~//IL ...".5040306070··T·.. '. "''15040706030Seeds Pods170 170160 160150 150140 140130 130OJ 120 QJ 120(f)0 0~E 110 ~E 110~ § 100 ~ § 100~ ~ 90 ~ ~ 90~- 80 ~- 80(,) (,)Time (Hours)_ Diabetic (Untreated)Treated with Petroleum Ether Extract (1 glkg)Treated with Chloroform Extract (1 glkg)Treated with Ethyl Acetate Extract (1 glkg)Time (Hours)Treated with Dichloromethane Extract (1 g/kg)Treated with Ammonical Chloroform Extract (1 glkg)Treated with Methanol Extract (1 glkg)Fig.1. E./Jed ofdi./Jerent chemical solvents extrads of P. speciosa on blood glw:ose levels in alloxan-diahetic raLs. Data are means ± SE (n = 4)162 PERTANlKAJ. TROP. AGRIC. SCI. VOL. 16 NO.3, 1993HYPOGLYCEMIC EFFECT OF EXTRACTS OF PETAl PAPANFig. 2. l!..jJect ofchlaroform extracts ofP. speciosa on normaland diabetic rats. Data are means ± SE (n = 4)F'ig.3. Dose-response relationship iffresh and ground seeds on bloodglucose levels ofaU.o:xan-<.lialx1icmls. Data are rneans±5E (n =4)24242020- ---~- --161612- .. - Diabetic 100 mglkg (48% Decreament)- . - Diabetic 250 mglkg (58% Decrement)- Diabetic 500 mglkg (77% Decrement)Time (Hours)12Diabetic (Untreated)SalineNormal (Seeds Extract 400 mg/kg)Normal (Pods Extract 400 mg/kg)Treated (Seeds Extract 400 mg/kg)Treated (Pods Extract 400 mglkg)----~:r--z.. ,:JI"--.,_. __ .... :s:...'w _- Diabetic (Untreated)--- SalineDiabetic 25 mglkg (24% Decrement)- - Diabetic 50 mglkg (31 % Decrement)Time (Hours)420,-----------------~400380360340320300 )C'~-.-i....~., /'-~-.-.-.- ..--.-.-.-.-.--.--.--280 ',,/./ \ x...,...._...1~ ~:~ ~""-L"/' \,o 220 I.. _t 200 's..""-r".._ ...:a-180160140~ ~~ . .:;.;.~-1:'~:£!::::::':;.~~~~.~.~.~~~.~.:~.~.~~.~.;~~~~~~80 -f---,.--,----,,--,----,,--,----,,--r----.--r-----.-----.j420 ,-----------=-----------,400380360340320300280E 260g 240220.[ 20018016014012010080 -t---,.--r----,--,----,--,--,--.----,,--,----,.--J(1 g/kg body weight) or the control (treatmentwith saline). Further work therefore concentratedonly on the chloroform fraction.Fig. 2 shows that there was insignifican tincrease in the blood glucose levels of normal ratsfed with 0.4 g ground seeds or pods together with1 g glucose/kg body weight. The normal rats hadan average blood glucose content of 124 mg/100ml, while the alloxan diabetic rats had an averageblood glucose level of 379 mg/100 ml after ingest-ing 1 g glucose/kg body weight.The blood glucose level of alloxan diabeticrats was reduced by 36±6 % to 288 mg/ 100 ml withthe oral treatment of 0.4 g/kg BW'pericarp (pod),and by 57±6 % to 236 mg/100 ml after the oraltreatment with 0.4 g/kg BW petai seed. The treat-ment could be seen to take effect within less thanan hour and lasted for at least 24 hours. The max-imum fall was observed 2 hours after oral adminis-tration. However, there was an initial rise in bloodglucose level between 0-3 hours, showing that theglucose was rapidly absorbed from the alimentarycanal and that the extract of petai took effect onlytwo hours after ingestion. The blood glucose levelof healthy and diabetic rats fed with saline plus1 g/kg BW glucose is shown for comparison.The seed had a higher activity than the p ri-carp. Fig. 3 shows the dose-response relationshipof petai seed on blood glucose level in diabeticrats. A dose of 25 mg/kg BW decreased theblood glucose by 24±4 %, yet a 4-fold increase indosage (100 mg/kg BW) only decreased theblood glucose by 43±5 %. Increasing the dosage20-fold (500 mg/kg BW) decreased the bloodglucose by 77±12 %. Further increasing thedosage (3 g seed/kg BW) decreased the bloodglucose level by 116±12 % i.e. bringing the glu-cose, level below that of a normal healthy rat.TABLE 1Reduction in blood glucose (%) ofdiabetic rats following the administration of chloroform extract of petai seedsdose hr 50 100 150 200 250 300 350 400 450 500(mg/kgbody wt)1 25.41 51.76 53.17 55.52 56.47 57.41 58.35 59.29 58.35 57.413 20.70 30.11 32.94 36.70 37.64 37.64 38.58 39.05 43.76 48.945 25.4 34.82 43.29 50.82 55.52 59.29 64 67.76 72 75.768 25.41 46.11 53.17 60.23 64 73.88 66.35 68.70 69.17 70.1111 13.17 30.11 36.70 44.23 47.05 49.41 51.76 53.64 44.70 36.7024 8.941 21.64 27.29 33.88 34.35 34.82 35.76 36.70 34.82 33.88PERTA IKA]. TROP. AGRIe. SCI. VOL. 16 NO.3, 1993 163FATHAlYAJAMALUDIN AND SUBAIu MOHAMED24201612__-- .. t:--a..... __ ..... __ ................... _. __ = -420 -,--------------------.,400380360340320300280E 260o 240~ 220.[ 20018016014012010080 +---,--....----,--....---,---,----,---,----,--....---,-----lThe percentage lowering of blood glucose atvarious doses of seed is presented in Table l.The optimum percentage lowering of blood glu-cose appeared to occur around 5-8 hours afteradministration of extracts regardless of the doselevel. Optimum dosage appeared to be around200 mg seed/kg body weight in the alloxan-induced diabetic rats. Except for the first 2hours, the response (percentage lowering ofblood glucose) appears to follow an exponentialrelationship to the dosage given with a high cor-relation coefficient of r2 = 0.99. The best fittedline for this correlationship is given as:y = 3.01 -Jx + 10.2where:y percentage lowering of blood glucose,(blood glucose level ofdiabetic rats - treated rats)x 100Time (Hours)Diabetic (Untreated)Saline25 mg/kg (No Decrement)50 mg/kg (18% Decrement)100 mg/kg (31% Decrement)250 mg/kg (47% Decrement)(blood glucose level ofdiabetic rats - healthy rats)and x = mg seeds/ kg body weight.Fig. 4. Dose-response relationship oj fresh and ground emptypods on blood glucose levels ojalloxan-diabetic rats. Dataare means ± SE (n = 4)Similarly the percentage lowering of blood glucoseat various doses of empty pod is presented by theequationy = 4.02 -Jx - 13where:y = % lowering of blood glucose,and x = mg pods/kg body weight.Except for the first 4 hours, the percentagelowering of blood glucose appears to follow anexponential relationship to the dosage given witha high correlation coefficient of r2 = 0.94. It cantherefore be generalised that for both the seedand pericarp, the response is approximately pro-portional to the square root of the dose given.The time taken for the pericarp to take effect andthe duration of the hypoglycemic activity areshown in Fig. 4. The pericarp had a lower activitythan the seed. At 25 mg/kg BW there was no sig-nificant activity. At 50 mg/kg BW the lowering ofblood glucose was 18±4 % and the activity at 100mg/kg and 250 mg/kg was 31±5 % and 47±5 %respectively. This is about half the activity of theseed.The fact that the blood glucose response topetai seeds and pods is square root to the dosemay indicate that the mechanism of action of theactive compounds in petai is peripheral. This isbased on comparison of dose response curves ofperipherally-acting compounds to centrally-acting ones (those causing the pancreas toincrease insulin production and release).Peripheral-acting compounds act directly on allthe cells in general, enabling more glucose toenter the cells. Chemical studies on the activecompounds of petai showed them to be sterols(results to be published) which can readily affectthe lipoprotein part of cell membranes. Furtherwork is being carried out to determine the mech-anism of action.Even though the activity of the pericarp(empty pod) and mesocarp (testa) are half ofthat from the seed, extraction of compoundsfrom the empty pod is viable because it consti-tutes 57 % of the whole pod and only the seedsare normally eaten while the outer skin is lesspalatable although edible. In Malaysia, cannedpetai seeds with anchovies and chilli sauce areavailable in the market. The empty pods aretherefore a waste product which can be used as araw material for the extraction of hypoglycemicmaterial.ACKNOWLEDGEMENTSThe authors are indebted to Mr Zainal AbidinJamin for valuable assistance; and MPKSN andIFS for funding the research.164 PERTANlKAJ. TROP. AGRIe. SCI. VOL. 16 NO.3, 1993HYPOGLYCEMIC EFFECT OF EXTRACTS OF PETAl PAPANREFERENCESGMEL! , R, R SUSILO and G.R FE!\TWICK. 1981.Cyclic polysulfides from Parkia speciosa.Phytochemistry 20(11): 251-253.HOLZMAN, G, R SUSILO and R GMEL! . 1982.Collisional activation study of cyclic polysul-fides. Org. Mass Spectrom. 17(4): 165-172.L NDQUIST, I. and C. RER PA. 1967 Blood glucoselevel in mice III. Corticotropin inducedhypogolycaemia. EuropeJ Pharmacol. 35: 2.SUHAILA MOHAMED, MOHAMED SHAMSUDDI ABDULRAHMAN, SABTURIAH SULAIMAN and FAUZIAHABDULLAH. 1987. Some nutritional and anti-nutritional components in jering (Pithecellobiumjeringa) , keredas (P. microcarpum) and petai(Parkia speciosa). Pertanika 10(1): 61-68.PA DEYA, S.N. 1972. Role of sulphides(thioethers) in biological systems. J Sci. Ind.Res. 31: 320-331.RILEY, V. 1960. Adaptation of orbital bleedingtechnic to rapid serial blood studies. Proc.Soc. Biol. Med. 104: 751.SUSILO, R. and RZ. GMEL! . 1982. Precursors ofcyclic polysulfides in seeds of Parkia speciosa.Naturforsch C. Biosci. 37C(7-8): 584-586.(Received 6January 1992)PERTANlKAJ. TROP. AGRIe. SCI. VOL. 16 NO.3, 1993 165

Hypoglycemic effect of extracts of Petai Papan (Parkia speciosa, Hassk)

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Hypoglycemic effect of extracts of Petai Papan (Parkia speciosa, Hassk)

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