Ir aq i J.Pharm.Sc i., Vol.15 (1 )  ,2006 10

A Study of the Furocoumarin Derivative of Ruta Chalepensis L. 
(Rutaceae) 

Ekbal H.AL-Khateeb, Ali A. AL-Shamma, Bajes A. Nehar 
Rece ived   2-3-2002      Accepted  23-10-2002 

ABSTRACT  
  The conte nt o f Furoc oumarin d eriv ativ es (Pso ra le ns ) of Ru ta  Ch alepe ns is  L. (Who le  p la nt) was  
stud ie d by s imp le  e xtra ctio n with p etro le um ethe r (b .p . 60 -8 0Co). The  res ults  indicate d that the  p la nt 
co ntains  a  total of a bo ut (0.01 5%). 
       Inves tigatio n of these  comp ounds  b y thin la yer chroma tograp hy (TLC) re vea le d the pres ence of at 
le as t four compounds  of whic h metho xs alen (8-metho xyp so ra le n) wa s is olated .     It wa s id entifie d and  
authentica te d with a  s ta nd ard by sp ec tral metho d, IR, NMR, Mas s sp ectra and HPLC.This  plant could  
be  c ons id ered  a s a go od so urce  for s up plying this co mpound , whic h is  wide ly use d in d ermatologic al 
prepa ra tio ns. 

  الخالصة
عماالته  ست العشاب با روف لدى متداولي ا مع كن متعددة و ما ي ا رع ف ي يز عراق والذ وجودة في ال ة الم ن النباتات الطبي سذاب م ان نبات ال

وري دراسـة     . الشعبية عراق٬ لذا فقد وجـد مـن الضـر عن النبات في ال ة  ي دراس ة وال توجد ا عن هذا النبات قليل ورة  ان الدراسات المنش
ونات النب كومـارين  مك رو مواد الفي من  هذا النبات  ما يحتويه  p( ات بصورة كيمياوية مفصلة والتعرف ع soralen (   طرق ال وفصـلها ـب

ة   .المتبع
ن     ري مـا وكو ور ى الفي وي عـل ة اثبتت لنا بان النبتة تحـت ولي ج اال p(ان النتائ so ra le n (        ذيبات ـم حسـن ال اء ا ة النتـق مل ا ـش ة  ال دراسـ م او ذا ـت ـل

م مكن استغ ة التي ي وي الوراق العض وفي ا ة  من جه ن الجذور  كل م ي  عيتها ف ن النبتة ثم تعيين كميتها ونو واد م خالص هذه الم ست ها ال ال
ى هة اخر ن من ج ة حرارة . والسيقا ي بدرج غل ر الذي ي وم ايث رولي ن البت رت النتائج با وقد اظه ة من النبات  مي مع ك م كان ° 80-°60فقد تم ج

ال ملة الستخ عضوية المستع سن المذيبات ال ومارين اح وك واد الفيور ps(ص م oralen .(    ة ريـق لقد اثبتت لنا نتائج فحـص المسـتخلص بط
التي  كا ميت  واد س هذه الم مركبات من  ة  ي النبات وقد تم فصل اربع كبات ف مر ود عدة  عن وج ة  ة الرقيق وكرافيا الطبق مات  ,BA1.:  كرو

BA 2, BA3, BA4 ركب االول ن الم خفيفة ) BA1(ا ورات  كل بل حرارة انصهاره تم فصله على ش ة  طويلة بيضاء درج 14و م ° 7-148
كومارين  رو كبات الفيو ن مر هو م ويدي و ركب االم كب اثبت بانه م مر هذا ال وي ل ميا كي pso(والتحليل ال ra le n(. ركب ما الم فقد تم ) BA2(ا

ة انصـهاره   رات بيضاء اللون درح ة الج      . م ° 78-76فصله على شكل بلو والصـيغ ي  جزيـئ وزن ال ين اـل عـي م ت د ـت مركـب    لـق هـذا ال ة ل زيئـي
حليل الكتلة  طة ت Ma(بواس ss  S pec tro sco py ( كب مر زيئي لل ن الج زيئية  BA2 (256(وان الوز ة الج C16H16Oوالصيغ ثم تم عمل . 3

NMR  وIR  ب مرك حاليل ان ال خالل هذه الت ن  جد م حيث و ركب ٬  شكل الكيمياوي لهذ الم كب لتحديد ال مر هذا ال يحتوي على ) BA2(ل
كو ورو ن نواة الفي ري p(ما so ra le n (جانبية ة  سل سل ه       . وترتبط هذه النواة ب كيـب د تر حدـي حاضـر لغـرض ت ت ال ي الوـق رر حفظه ـف لذا فقد تق

ريب مستقبل الق ب .الكيمياوي في ال مرك ها ) BA4( و ) BA3 (اما ال غرض دراست ر ل ها في الوقت الحاض هما قليلة وتقرر حفظ ميت كانت ك
ستقبل رة مفصلة اكثر بالم   . بصو

  
 
 
INTRODUCTION  
  Ruta  Cha le pensis L. (Rutace ae ) is a cultivate c 
plant [1] wid ely grown a s ga rd en o rname ntals in 
Baghda d a nd its v ic inity. Ruta, Rue  and 
Sad ha b are common names  fo r this plant. 
Rutac ea e (a ls o te rme d Rue family) is  o ne  of 
the imp orta nt familie s, tha t comp ris es many 
ge nera and sp ec ie s which have  bee n us ed as  a 
so urce o f d rugs a nd  medicine s [2]. The 
de coc tion o f this p la nt has  b een use d in folk 
re med y as  c arminative to cure s to mac h ache in 
children[3]. 
This  study was  c arried in connec tio n with a 
program of using the  chemica l co ns titutes  of 
Iraq i Me dicina l P la nts a s a  pote ntia l for 
industrial purpo se s. Literature s urvey re vea le d 
that Ruta  c ha le penis  L., c ontains  many 
importa nt c ompo ne nts[3-9], among w hich are 
co umarins  a nd Furo co umarin de rivative s.Since 

no prev io us work has been done  on this 
Iraq i p la nt was  found that stud y o f these  
comp ounds , is of adv anta ge for its ec ono mic al 
value  a nd  use s in d ermatologic al pre pa ra tions, 
as a vailable impo rted  one s are ve ry e xp ens iv e. 
MATERIALS and METHOD  
 The p la nt mate rial was  co llec te d during 
Sep te mbe r and Octob er 1 997  from the ga rd ens  
of the  v ic inity o f the  c ity of Baghda d. F our and 
a half kilograms  o f po wdered  plant mate rial 
(a eria l plants  and  ro ots) we re  extracte d with 12  
liters of p etro le um ethe r (b.p. 6 0-80 Co), under 
re flex fo r 7 2 hours. The p etro le um e ther 
extract was  filtered  and  e vap orated  to drynes s, 
und er reduce d p re ss ure at a  temp erature not 
exc ee ding 40  C o, le aving a  d ark gree n re sidue  
(8 0 gm). 

 
  
Dep artmen t of Pha rmco gna cy,Co lleg e of Phar macy, Un iver sity of Ba ghd ad, Bag hda d-I ra q 
 
 



Ir aq i J.Pharm.Sc i., Vol.15 (1 )  ,2006 11

Separation and Fractionation of 
Furoc oumarin Derivatives: 
    Different p artitio n proce ss es toge ther with 
co lumn chroma tograp hy technique w ere trie d 
but with uns ucc es sful re sults. So the  s ep aratio n 
and frac tionation wa s ca rried b y us ing 
prepa ra tiv e this  la yer chroma tograp hy. 
  A qua ntity of 20  gm of the re sidue  wa s 
diss olved  in a minimum a mount of chloroform 
and a pplied toge ther with s tand ard p so ra le n 
re fe re nc e comp ounds  (metho xs alen and 
imperatorin) on a  numb er o f prepa ra tive  TLC 
plates  o f s ilic a ge l GF 254  (0.75  mm thickne ss) 
and d ev elope d with so lv ent sys tem benze ne-
ac etone  (90 :1 0 v/v). 
     The d eve lope d plate s we re  v is ua lize d 
under U.V light (3 66 nm wave length). F our 
major bands we re  o bse rv ed  a nd de signated  a s 
ba nd I, II, III and IV. 
  Ba nd II and  III were o verlapp ed , and  b oth 
ha ve very c lo se  Rf va lues  to  that o f the 
standard reference me thoxs alen. 
   The se  two  b and s were se pa ra te d by multiple 
de velop ment us ing the sa me so lv ent system to 
give  two co mpo und s. 
    Band  I ga ve c omp ound I as  ye llowis h 
crys ta ls  (25  mg) up on re crys ta llization out of 
boiling metha nol, a fluffy c olorle ss  crys tals 
was  o btained  (20  mg) ha ving a s harp melting 
point of 1 47.5o. Out of ba nd  II, comp ound 2 
was  iso la te d as ye llowish p risma tic  crys tals 
(1 25 mg), up on re crystallization out o f hot 
pe troleum ether (b.p. 60 -8 0 Co) a colorles s 
lo ng p risma tic  c rystals (1 20  mg) was  o btaine d 
ha ving a s harp me lting po int of 78  Co. 
   Out of b and III, compound 3 was  is olated a s 
a yellowish crys tals (2 .7  mg) upo n 
re crys ta llization out of bo iling etha no l fluffy 
like  crysta ls  (2.5 mg) was  o btaine d. 
Out of b and IV, compound 4  wa s isolate d as a n 
oily material. The total yields  of thes e 
co mpounds  is olated fro m the  total extrac t (8 0 
gm)– were: 8 0 mg of c omp ound 1; 4 80 mg of 
co mpound 2; 10 mg of c omp ound 3; a nd oily 
material d es ignated  as co mpo und  4  . 

 ESULTS and DISCUSSION R 
   Out o f thes e co mpounds , only c omp ound 1 
was  ide ntified and c onfirmed to b e 
metho xs alen (xanthotoxin) a s it ha s similar 
melting point, CH analys is , and R f va lues in 
different s olvent s ys te ms with s tand ard 
re fe re nc e of me thoxsa le n. 
     An as sa y b y HPLC also was done for 
co mpound 1, us ing metho xsa le n U.S.P. 1 % a s 
a standard re fe re nc e. The re te ntion time  wa s 
exactly the s ame , furthe r id entifica tion wa s 
ca rrie d out by enric hment tec hnique, in whic h 
the reference  was a dde d ind iv id ually to 
co mpound 1, us ing three  different mobile 
phase s (Fig. 1 a, b, c). 

Other s pec tra l metho ds were us ed  for further 
confirmatio n. The  ma ss  s pe ctrum o f c ompo und  
1 indicates  a  mo le cula r io n pea k at m/e  21 6. 
The  fra gme ntation pa ttern showing s ignifica nt 
pea ks  a s shown in (Fig. 2 ). The  NMR s pe ctrum 
in CDCl3 (Fig. 3 ) co nfirmed  the  pres ence of 
eight protons. The proton (H4) a nd(H5)   a s tw o 
doublet a t δ  6 .8  (H4) and δ 7.71 (H5) (s hielded  
(J ) is sma ll as  e xpe cted  for the furo r ring (J =1-
2 Cps )). (H3) pro to n showed  s inglet pea k at δ 
7.82 (a ro matic p ro to n) (H1) and  (H2) protons  
also a ppe ared a s tw o doublet pea ks  a t δ  7 .3 5, δ 
7.26 for (H1) pro to n, a t δ  6.43  a nd  δ 6.31  for 
(H2) p roton. Me thoxy group  p ro to ns s howe d at 
δ 4.3. The IR spe ctrum (KBr) (Fig. 4) c onfirms  
the structure. C = O s tre tc hing ba nd at 1 575  
cm-1 fo r the α1 β-uns aturated  lacto ne. Tw o 
bands  at 1 600  cm-1 157 5 c m-1 fo r (C = C) 
stre tc hing v ib ra tion o f the aroma tic ring. (C = 
C) alefinic ba nd a t 16 60 cm-1 [10]. Two ba nd s at 
145 0 cm-1 and 1 375  cm-1 for as ymme tric  
bending vibration of CH3 group res pe ctiv ely. 
The  s tretc hing (C-O-CH3) a ppe ars at 12 80 cm

-1 
fo r as ymme trical stre tc hing a nd 10 20 cm-1 for 
symmetrica l s tre tc hing. Two  characteristic  
stre tc hing vibratio ns  fo r the e ster func tion 
app ear at 1 145  c m-1 and 109 0 cm-1. The  mo ot 
cha ra cteris tic  out of plane C-H be nding 
vibra tio n at 8 22 cm-1 a nd  7 60 cm-1. Moreo ver 
the C-H o ut p la ne  b end ing vibration at 87 0 cm-
1 is due to  o ne fre e (H) in benze ne ring. F inally 
the o ut o f plane  o le finic (H) (ciso le fe in) is  
prese nt at 7 55 cm-1 in a gree ment with the  
structure [10]. 
   All the  abo ve mentione d s pec tral da ta  
confirmed that c ompo und 1  is  me thoxsalen 
hav ing the follo wing structure: 
 
 
 
 
 
 
 
 

Co mpound 2, 3, and  4 le ft fo r furthe r 
identific ation studies 

 
 
 
 
 
 
 
 
 
 
 
 
 

OCH3

O O O



Ir aq i J.Pharm.Sc i., Vol.15 (1 )  ,2006 12

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 

 
 
 
 
 

 
 

 
 
 
 

Fig (1 a )H.P.L.C for a mixture  o f e qua l quant itie s  o f the  co mpo und (BA1) and the  s tandard 
re fe re nce Oxsoralon : (Me tho xsa le n U.S.P 1% ) with s eco nd mob ile  phase 

 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 
 
 
 

 
 
 

 
 
 
 
 
 

 
 
 
 
 

 
 
 

 
 

Fig (1b) HPLC fo r the  co mpound (BA1) compare d to the  standard re fe re nce 
oxso rale n ;(Me thoxsale n u.s.p 1 %)

  



Ir aq i J.Pharm.Sc i., Vol.15 (1 )  ,2006 13

 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 

 
 
 
 

 
 
 
 

Fig .(1c ) HPLC for a  mixture  of e qual quantitie s of the  compound (BA1) a nd the  standard 
re fe re nce  Oxso ra le n ; (Me thoxsale n C.S .P 1 % )  wit h third mo bile  pha se  

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

  
Fig (2) Mass Spec trum of the  compo und (BA1) 

 
 
 
 



Ir aq i J.Pharm.Sc i., Vol.15 (1 )  ,2006 14

Fig  (3): NMR Spe ctrum o f the  co mpound (Ba1) de te rmine d in CDCL3 

 
Fig (4 ) :IR Spe ctrum o f co mpou nd (BA1)

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 



Ir aq i J.Pharm.Sc i., Vol.15 (1 )  ,2006 15

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