Identification by HPLC-DAD and HPLC-MS Analyses and

Quantification of Constituents of Fennel Teas and Decoctions

Anna Ri ta Bi l i a,* Margheri ta Fumarol a, Sandra Gal l ori , Gi ovanni Mazzi , and F. F. Vi nci eri
Di parti mento di Sci enze Farmaceuti che, Uni versi ty of Fl orence, vi a Gi no Capponi 9, 50121 Fl orence, I tal y
Qual i tati ve and quanti tati ve di fferences among the consti tuents i n vari ous fennel (Foeniculum
vulgare Mi l l ., fami l y Api aceae) teas prepared by cl assi cal i nfusi on, mi crowave decocti on, and
di ssol uti on are reported. Di fferent commerci al starti ng materi al s, such as frui t (unbroken and
crushed), four herbal teas, and two i nstant herbal teas were eval uated. Chl orogeni c aci d (1),
querceti n-3-O--D-gl ucuroni de (2), p-ani sal dehyde (3), and trans-anethol e (4) were i denti fi ed by
HPLC-DAD and HPLC-MS as consti tuents of fennel teas. No coumari ns, whi ch are characteri sti c
consti tuents of pl ants of Api aceae fami l y, were found. Trans-anethol e (4), the mai n consti tuent of
the essenti al oi l , was present i n al l teas. I n addi ti on p-ani sal dehyde (3), a degradati on product of
trans-anethol e, was al so i denti fi ed i n al l teas wi th the excepti on of two sampl es. Chl orogeni c aci d
(1) and querceti n-3-O--D-gl ucuroni de (2) were al so present i n al l teas. I n addi ti on, mi nor
uni denti fi ed fl avonol consti tuents were found i n two teas. Qual i ty, acti vi ty, and safety of the content
of the i nvesti gated preparati ons are al so di scussed.
Keywords: HPLC-DAD and HPLC-MS; fennel; herbal teas and instant herbal teas; infusion
and microwavedecoction; essential oil; polyphenols
I NTRODUCTI ON
Many herbal drugs whose effi cacy has been attri buted
to the essenti al oi l content are used as teas, i .e., cham-
omi l e, peppermi nt l eaf, ani seed, and fennel (Czygan,
1989). However, very l i ttl e i s known about the qual i ta-
ti ve and quanti tati ve composi ti on of the essenti al oi l
andother consti tuents present i n the teas. Furthermore,
chi l dren and newborns are frequent consumers of these
teas, so that knowl edge of thei r consti tuents represents
a very i mportant step for the eval uati on of thei r safety
and effi cacy.
I n thi s work the consti tuents of teas from fennel were
eval uated. Thi s drug i s descri bed i n the 2nd edi ti on of
the European Pharmacopoei a both as Sweet Fennel
and Bi tter Fennel , whi ch consi st of the dry, whol e
cremocarps and meri carps of Foeniculumvulgaresub.
vul gare, var. dul ce (Mi l l er) Thel l ung and Foeniculum
vulgaresub. vul gare, var. vul gare Mi l l er, respecti vel y.
The sweet vari ety i s mai nl y marketed i n I tal y, so i t was
empl oyed for our i nvesti gati on. Fennel and i ts herbal
drug preparati ons are used for dyspepti c compl ai nts
such as mi l d, spasmodi c gastri c-i ntesti nal compl ai nts,
bl oati ng, and fl atul ence. I t i s al so used for the catarrh
of the upper respi ratory tract (Czygan, 1989; Madaus,
1976; Merkes, 1980; Forster et al ., 1980; Forster, 1983;
Wei b, 1991; Reynol ds, 1993). Accordi ng to the 9th
edi ti on of the German Pharmacopoei a, the essenti al oi l
i s currentl y bei ng consi dered as responsi bl e for the
pharmacol ogi cal properti es of fennel but i t cannot be
gi ven as such to chi l dren and newborns because of i ts
ri sk of l ari ngospasms, dysponeas, and states of agi tati on
(Dorsch et al ., 1993). I n thi s study, teas from commerci al
frui t were obtai ned from both crushed frui ts, accordi ng
to the publ i shed recommendati ons (Czygan, 1989), and
from unbroken frui ts. I n addi ti on, teas obtai ned from
prepackaged teabags and from freeze-dri ed products of
fennel have al so been i nvesti gated. Prepackaged teabags
marketed i n I tal y contai n unbroken and/or crushed frui t
or powdered drug. However, the use of unbroken frui t
to prepare i nfusi ons i s i ncorrect. I ndeed some di ffi cul ti es
exi st i n extracti ng the essenti al oi l by i nfusi on due to
i ts i ntracel l ul ar l ocal i zati on. However, crushed or pow-
ered frui t gradual l y l ose thei r essenti al oi l content
duri ng agi ng (Czygan,1989). Fi nal l y, the qual i tati ve and
quanti tati ve contents of the teas prepared usi ng pre-
packaged teabags 7 and 30 days after openi ng were al so
eval uated to i nvesti gate the possi bl e l oss of vol ati l e
consti tuents over ti me. Accordi ng to the 2nd edi ti on of
the European Pharmacopoei a monograph, sweet fennel
contai ns not l ess than 2.0% v/m of essenti al oi l , cal cu-
l ated wi th reference to the anhydrous drug. The es-
senti al oi l i s consti tuted mai nl y by anethol e (80%), and
i t contai ns not more than 10% estragol e and not more
than 7.5% fenchone (Brand, 1993). Other mi nor con-
sti tuents may be present i ncl udi ng R-pi nene, l i monene,
-pi nene, -myrcene, and p-cymene (Brand, 1993; Toth,
1967; Trenkl e, 1972). Furthermore, sweet fennel con-
tai ns other nonvol ati l e consti tuents such as fl avonoi ds
and coumari ns (Kunzemann andHerrmann, 1977; Mur-
ray et al ., 1982) whi ch have not recei ved much attenti on
wi th regard to pharmacol ogi cal properti es. However,
thei r presence i n the teas coul d contri bute to the
pharmacol ogi cal acti vi ty of such herbal drug prepara-
ti ons.
EXPERI MENTAL PROCEDURES
Solvents. Acetoni tri l e was HPLC grade from Merck (Darm-
stadt, Germany); 85%formi c aci d was provi ded by Carl o Erba
(Mi l an, I tal y). Water was puri fi ed by a Mi l l i -Qpl us system from
Mi l l i pore (Mi l ford, MA).
Standards. I ndena Research Laboratori es (Settal a, Mi l an,
I tal y) ki ndl y provi ded ruti n (batch n. K12408717, standard
* Correspondi ng author. Phone: +390552757288. Fax:
+39055240776. E-mai l : bi l i a@farmfi .sci farm.uni fi .i t.
4734 J. Agric. Food Chem. 2000, 48, 47344738
10.1021/jf000171n CCC: $19.00 2000 American Chemical Society
Published on Web 08/08/2000
puri ty 88.17% consi deri ng the content of resi dual sol vents,
moi sture, and amount of i mpuri ti es). Chl orogeni c aci d was
purchased from Extrasynthese (Genay, France). Trans-ane-
thol e and p-ani sal dehyde were purchased from Si gma Chemi -
cal s (Mi l an, I tal y). Querceti n-3-O--D-gl ucuroni de was i sol ated
and characteri zed by the authors. The puri ty val ue of these
standards was cal cul ated or gi ven by the suppl i ers (89.3, 99.0,
98.0 and 90.5%, respecti vel y).
Samples. Aboca S.p.A. (Sansepol cro, Arezzo, I tal y) ki ndl y
offered commerci al sampl es of frui ts of Foeniculum vulgare,
subsp. vulgare var. dul ci s (Mi l l er) Thel l ung. The drug (l ot
59710, 11/97) contai ned 27.7 mL/kg of essenti al oi l determi ned
accordi ng to the 2nd edi ti on of European Pharmacopoei a assay.
Herbal teas and i nstant herbal teas l i sted i n Tabl e 1 were
purchased from l ocal pharmaci es and grocery stores and
named as products A-F.
Sample preparations. Tradi ti onal teas were prepared
from frui ts by i nfusi on of 2.5 g of both unbroken and freshl y
crushed drug i n 150 mL of i nci pi ent boi l i ng water, l eavi ng i t
for 20 mi n i n a covered cup and occasi onal l y sti rri ng (Czygan,
1989; 9
th
edi ti on of the German Pharmacopoei a). Then the
suspensi ons were rapi dl y fi l tered through cotton wool . A 5-mL
fracti on of the fi l trate was used as such for the HPLC anal ysi s.
Teas were al so prepared by 2 mi n decocti on from 2.5 g of both
unbroken and crushed frui ts i n a covered cup contai ni ng 150
mL of water i n a mi crowave (potency 600 W). The suspensi ons
were rapi dl y fi l tered and processed as descri bed for the
tradi ti onal teas. I n addi ti on, tradi ti onal teas were obtai ned
usi ng teabags of products A-D and named Ac, Bc, Cc and Dc,
respecti vel y. These were processed as previ ousl y descri bed for
the other sampl es. Other teas (Am, Bm, Cm, and Dm) were
prepared by decocti on i n mi crowave of a teabag of products
A-D and after removi ng the teabags, were processed as
al ready reported. I n addi ti on, teas al so were obtai ned by
di ssol vi ng two i nstant herbal teas (E and F) i n warm (30 C)
water (150 mL) usi ng 7 g and 1 g of powders respecti vel y,
accordi ng to the l abel suggesti ons and named Ed and Fd.
These teas were processed i n the same manner as the other
sampl es.
HPLC-DAD AnalysisInstrumentation. The HPLC sys-
tem consi sted of a HP 1090L i nstrument equi pped wi th a di ode
array detector (DAD) and managed by a HP 9000 workstati on
(Hewl ett-Packard, Pal o Al to, CA). The col umn was a Li -
Chrosorb RP18 (5m, 250 4 mm i .d.) (Merck, Darmastadt,
Germany) mai ntai ned at 26 C and equi pped wi th a precol umn
Li Chrosorb RP18 (5m, 10 4 mm i .d.) (Merck, Darmastadt,
Germany). The mobi l e phase was a four-step l i near sol vent
gradi ent CH
3CN/H2O (see Tabl e 2) wi th HCOOH (pH ) 3.2)
duri ng a 40-mi n peri od at a fl ow rate of 1 mL/mi n. The i njected
vol ume of sampl e was 10 L of sol uti on. UV-vi s spectra were
recorded i n the range 190-450 nm, and chromatograms were
acqui red at 210, 254, 280, and 330 nm. Peaks were detected
at 254 nm. A typi cal chromatogram i s reported i n Fi gure 1.
The t
R val ues for chl orogeni c aci d (1), querceti n-3-O--D-
gl ucuroni de (2), p-ani sal dehyde (3), and trans-anethol e (4)
were 8.07, 18.8, 25.56, and 35.93 mi n, respecti vel y. Structures
of compounds 1-4 are reported i n Fi gure 2.
HPLC-MS Analysis Instrumentation. The HPLC sys-
tem previ ousl y descri bed was i nterfaced wi th a HP 1100 MSD
API -el ectrospray (Hewl ett-Packard, Pal o Al to, CA). The i n-
terface geometry, wi th an orthogonal posi ti on of the nebul i zer
wi th respect to the capi l l ary i nl et, al l owed the use of anal yti cal
condi ti ons si mi l ar to those of the HPLC-DAD anal yses. The
same col umn, mobi l e phase, ti me peri od, and fl ow rate were
used. Mass spectrometry operati ng condi ti ons were opti mi zed
i n order to achi eve maxi mum sensi ti vi ty val ues: gas temper-
ature 350 C at a fl ow rate of 10 L/mi n, nebul i zer pressure 30
psi , quadrupol e temperature 30 C, and capi l l ary vol tage 3500
V. Ful l scan spectra from m/z 100 to 800 i n the negati ve and
Table 1. Investigated Herbal Teas and Instant Herbal
Teas
test
product
type of
preparati on
recommended
dosages (g) l ot content
A teabags 2.3 08141 brown powder
B teabags 2.0 11H unbroken and
crushed frui ts
C teabags 2.0 309 fi nel y ground
pl ant materi al
D teabags 2.25 31 unbroken frui ts
E powder 1.0 freeze-dri ed
extract
F powder 7.0 freeze-dri ed
extract
Figure1. Chromatographi c profi l e of the conventi onal tea of
freshl y crushed frui ts wi th the HPLC-MS attri buti ons of the
components detected: 1, chl orogeni c aci d; 2, querceti n-3-O-
gl ucuroni de; 3, p-ani sal dehyde; 4, trans-anethol e.
Figure 2. Chemi cal structures of compounds 1-4.
Table 2. Mobile-Phase Composition Used for the
HPLC-DAD Analysis
ti me (mi n) % H2O % CH3CN fl ow (mL/mi n)
0.10 88.0 12.0 1.00
10.00 82.0 18.0 1.00
15.00 82.0 18.0 1.00
30.00 55.0 45.0 1.00
35.00 0.0 100.0 1.00
42.00 0.0 100.0 1.00
50.00 88.0 12.0 1.00
Identification of Constituents of Fennel Teas J. Agric. Food Chem., Vol. 48, No. 10, 2000 4735
posi ti ve i on modes were obtai ned (scan ti me 1 s). The i njected
vol ume of sampl e sol uti on was 10 L.
Identification of Tea Constituents. I denti fi cati on of
consti tuents 1-4 was performed by HPLC-MS anal ysi s and/
or by compari ng the retenti on ti me of the peaks wi th authenti c
sampl es. The puri ty of peaks was checked by compari ng the
UV spectra obtai ned wi th a DAD coupl ed to the HPLC system
wi th those obtai ned by authenti c reference sampl es and/or by
exami nati on of the MS spectra.
Linearity. Li neari ty range of responses was determi ned on
fi ve concentrati on l evel s wi th three i njecti ons for each l evel .
Cal i brati on graphs for HPLC were recorded wi th sampl e
amounts rangi ng from 0.10 to 2 g (r > 0.999).
Repeatability. To eval uate the repeatabi l i ty, si x fennel
frui t sampl es from the same batch were crushed and i m-
medi atel y used to prepare teas that were anal yzed as such by
RP-HPLC. Each consti tuent of such fennel teas was eval uated
to cal cul ate the rel ati ve standard devi ati on. The fol l owi ng data
were obtai ned: chl orogeni c aci d 1.93%, ruti n 1.85%, querceti n-
3-O--D-gl ucuroni de 1.96%, p-ani sal dehyde 2.09%, and trans-
anethol e 2.27%.
Reproducibility. To eval uate the reproduci bi l i ty of the
i njecti on i ntegrati on, 10 L of a standard sol uti on of ruti n (0.1
g/1 L) and of crushed fennel frui t sampl e of tradi ti onal tea
preparati on were i njected si x ti mes and the rel ati ve standard
devi ati on val ues were cal cul ated. The fol l owi ng data were
obtai ned: chl orogeni c aci d 1.32%, ruti n 0.80%, querceti n-3-
O--D-gl ucuroni de 0.98%, p-ani sal dehyde 1.12%, and trans-
anethol e 1.27%.
Quantitation. Al l the fennel teas were anal yzed i n tri pl i -
cate and a cal i brati on graph wi th si x datapoi nts of external
standard was used. The contents of consti tuents were cal cu-
l ated taki ng i nto account the mean of the response factor of
ruti n i n the reference sol uti ons, i .e., area/concentrati on (mg/
mL) puri ty/100, and the response factor of the consi dered
consti tuent rel ati ve to ruti n (RRF), as reported i n the l i terature
(Brol i s et al ., 1998). Thi s val ue was determi ned by cal cul ati ng
the rati o between the average response factor of each com-
pound and the average response factor of ruti n at 254 nm.
RESULTS AND DI SCUSSI ON
Thi s i s the fi rst report of the anal ysi s of the consti tu-
ents of fennel teas usi ng a si mpl e, di rect, rapi d, and
robust RP-HPLC method. Accuracy, reproduci bi l i ty
and speed were found acceptabl e. Fennel teas were
prepared both by i nfusi on wi th occasi onal sti rri ng
accordi ng to the 9
th
edi ti on of German Pharmacopoei a,
and by decocti on i n mi crowave as descri bed i n the
Experi mental Secti on. Commerci al frui ts of Foeniculum
vulgare, subsp. vulgare, var. dul ci s (Mi l l er) Thel l ung
contai ni ng 27.7 mL/Kg of essenti al oi l , commerci al
teabags, and i nstant teas were used i n thi s i nvesti ga-
ti on.
Four major consti tuents were i denti fi ed: chl orogeni c
aci d (1), querceti n-3-O--D-gl ucuroni de (2), p-ani sal de-
hyde (3) and trans-anethol e (4). A of 254 nm was used
for HPLC quanti tati ve eval uati on of consti tuents, as al l
the consti tuents showed appreci abl e absorbance at thi s
wavel ength. Ruti n was used as external standard. Good
l i neari ty of the cal i brati on curves was achi eved between
0.1 and 2 g (r > 0.999); the repeatabi l i ty and reproduc-
i bi l i ty of the method showed sati sfactory resul ts.
Al l compounds were i denti fi ed by means of HPLC-
DAD and HPLC-MS anal yses. Peak 1i n the chromato-
gram of Fi gure 1 di spl ays UV spectra wi th maxi ma at
236 and 327 nm and shows a quasi -mol ecul ar i on
[M+H]
+
at m/z 355 and [M+H-H
2
O]
+
i on at m/z 337.
Peak 1was i denti fi ed as chl orogeni c aci d by compari son
of t
R
, UV spectra, and MS spectra wi th those of an
authenti c sampl e. Peak 2 di spl ays UV spectra wi th
maxi ma at 259 and 355 nm, typi cal of fl avonol s.
Moreover, the presence i n the ESI -MS spectrum of an
i on at m/z 301 i ndi cates that thi s compound i s a
querceti n deri vati ve. A quasi -mol ecul ar i on [M-H]
+
at
m/z 477 and the fragment i on at m/z 301 are evi dent
due to the l oss of gl ucuroni c aci d. Peak 2was i denti fi ed
as querceti n-3-O--D-gl ucuroni de by compari son of t
R
,
UV spectra, and MS spectra wi th those of an authenti c
sampl e. Peaks 3and 4were i denti fi ed as p-ani sal dehyde
and trans-anethol e by compari son of t
R
and UV spectra
(maxi ma at 281 and 257 nm, respecti vel y) and MS
spectra wi th those of authenti c sampl es. P-ani sal dehyde
shows a quasi -mol ecul ar i on [M+H]
+
at m/z 137 and
trans-anethol e shows a quasi -mol ecul ar i on [M+H]
+
at
m/z 149.
Trans-anethol e (4), the mai n consti tuent of essenti al
oi l , was present i n al l teas. I n addi ti on p-ani sal dehyde
(3), a degradati on product of trans-anethol e, not present
i n fennel essenti al oi l obtai ned by steam di sti l l ati on, was
al so i denti fi ed i n al l teas wi th the excepti on of those
from products A and E. Chl orogeni c aci d (1) and
querceti n-3-O--D-gl ucuroni de (2) were al so present i n
al l teas i n addi ti on to mi nor uni denti fi ed fl avonol
consti tuents found i n teas obtai ned from products A and
C. For al l the i nvesti gated teas the content of the
essenti al oi l per 150 mL of preparati on was cal cul ated
and the amounts of consti tuents 1-4, usi ng the re-
sponse factor rel ati ve to ruti n (RRF), were al so reported.
The resul ts are gi ven i n Tabl es 3 and 4.
The qual i tati ve and quanti tati ve composi ti on of vol a-
ti l e consti tuents i n the vari ous teas was qui te di fferent.
I n tradi ti onal teas of freshl y crushed frui ts 88% of the
vol ati l e consti tuents consi sted of trans-anethol e (4),
whi ch was present i n an amount about 8-fol d more than
p-ani sal dehyde (3). However, there was a hi gher content
of p-ani sal dehyde i n the teas obtai ned by mi crowave
decocti on, probabl y due to the di fferent extracti on
condi ti ons and/or to degradati on of 4to 3. I f unbroken
frui ts were used, the content of p-ani sal dehyde was
hi gher, as wel l . A tea obtai ned by di ssol uti on (Fd)
showed hi gh content of trans-anethol e and a smal l
amount of p-ani sal dehyde, and thei r percentages were
si mi l ar to those of tradi ti onal teas of freshl y crushed
Table 3. List and Relative Amounts
a
of Constituents Detected in Teas fromFruits and in Instant Herbal Teas
teas
unbroken frui ts
(conventi onal )
unbroken frui ts
(mi crowave)
crushed frui ts
(conventi onal )
crushed frui ts
(mi crowave) Ed
b
Fd
b
used dose (g) 2.5 2.5 2.5 2.5 1.0 7.0
chl orogeni c aci d (mg) 1.4 ( 0.11 1.6 ( 0.19 1.7 ( 0.21 2.0 ( 0.20 0.5(0.09 1.4(0.12
querceti n-3-O--D-
gl ucuroni de (mg)
6.9 ( 0.08 7.4 ( 0.08 8.6 ( 0.10 10.2 ( 0.11 0.3(0.08 2.4(0.27
other fl avonol s(mg) Not found Not found Not found Not found Not found Not found
anethol e (mg) 2.6 ( 0.21 1.6 ( 0.19 10.0 ( 0.11 4.3 ( 0.39 5.7(0.44 9.1(0.81
ani sal dehyde (mg) 1.3 ( 0.11 1.4 ( 0.13 1.1 ( 0.16 2.2 ( 0.20 Not found 0.3(0.09
a
Means ( SD of three repl i cates.
b
Di ssol ved i n warm water.
4736 J. Agric. Food Chem., Vol. 48, No. 10, 2000 Bilia et al.
frui ts. The other sampl e obtai ned by di ssol uti on (Ed)
showed onl y trans-anethol e. Al so, i n conventi onal teas
obtai ned by the i nfusi on of teabags trans-anethol e
content was hi gher than that obtai ned by mi crowave
decocti on. Thi s val ue sl i ghtl y decreased i n teas prepared
usi ng teabags 7 or 30 days after package openi ng (see
Tabl e 4). Both teas (Cc and Cm) obtai ned by product C
showed the presence of onl y p-ani sal dehyde.
Concerni ng the other substances found i n the teas,
chl orogeni c aci d (1) i s a very common metabol i te of
hi gher pl ants, and querceti n-3-O--D-gl ucuroni de (2) i s
a qui te unusual gl ycosi de of querceti n whi ch, however,
i s a known consti tuent of fennel frui t (Kunzemann and
Herrmann, 1977). Chromatograms of Ac, Am, Cc, and
Cm teas showed traces of other fl avonoi ds. Thei r
maxi ma i n the UV spectra were typi cal of fl avonol
deri vati ves, probabl y gl ycosi des of querceti n and/or
kaempferol . These compounds, however, are al ready
reported as mi nor consti tuents of aeri al parts of Foen-
iculumvulgare(Kunzemann and Herrmann, 1977). I n
al l teas they were detected i n very l ow concentrati ons
and i t was not possi bl e to ful l y i denti fy them. Thei r
i sol ati on and i denti fi cati on i s presentl y i n progress.
The contents of chl orogeni c aci d (1) and querceti n-3-
O--D-gl ucuroni de (2) were al ways hi gher i n the teas
obtai ned by mi crowave decocti ons than those obtai ned
by i nfusi on, probabl y due to the extracti on methodol ogy.
The content of 1 per cup i n tradi ti onal teas of freshl y
crushed frui ts was 1.7 mg, whi ch i ncreased to 2.0 mg
i n teas from mi crowave decocti on of freshl y crushed
frui ts. The content of 2was 8.6 mg per cup i n tradi ti onal
teas of freshl y crushed frui ts and 10.2 mg i n teas
obtai ned by decocti on usi ng a mi crowave. The content
of these pol yphenol s was sl i ghtl y l ower i n teas obtai ned
from unbroken frui ts (see Tabl e 3). Tea Ed showed a
very l ow content of these substances (0.5 mg of 1 and
0.3 mg of 2), whereas Fd showed a hi gher content (1.4
mg of 1 and 2.4 mg of 2). The amount of 1 i n Fd was
general l y si mi l ar to that i n al l teas obtai ned from the
other products i nvesti gated (A-D). Al l teas obtai ned
from products A-D showed a content of 2si mi l ar to that
obtai ned i n the tea from di ssol uti on of product F, or
more (val ues between 1.2 mg and 6.3 mg). However, i n
every case these val ues were l ower than those found i n
tradi ti onal teas of freshl y crushed frui ts. I n general , the
consi derabl e amount, per cup, of these two pol yphenol s
coul d suggest thei r contri buti on i n the acti vi ty of these
herbal drug preparati ons.
Coumari ns, whi ch are characteri sti c metabol i tes of
fennel and i n general of the Api aceae fami l y (Murray
et al ., 1982), were found i n none of the teas i nvesti gated.
Thi s fact was very i mportant for drug use safety.
Concerni ng fennel anti -spasmodi c effects, i t i s re-
ported that i n vi vo studi es of 2-3 g of tradi ti onal teas
of freshl y crushed frui ts/kg body wei ght i n cats i nhi bi ted
i n si tu i l eum spasm (approxi matel y 150 s, 50%) i nduced
by acethyl col i ne and hi stami ne (Schuster, 1971). There-
fore, onl y Fd, whi ch shows a content of vol ati l e con-
sti tuents i n thi s range, may be consi dered acti ve for thi s
therapeuti c i ndi cati on. Concerni ng fennel secretol yti c
and expectorant effects, i n vi vo studi es on rats refer onl y
to anethol e. I nhal ati on of anethol e (3 mg/kg) augmented
the vol ume output of respi ratory tract fl ui d (Boyd and
Sheppard, 1971). I f the necessary dosage for an adul t
human of 70 kg was consi dered, the expected anethol e
dosage woul d be 210 mg of anethol e correspondi ng to
about 235 mg of essenti al oi l . Si nce i t i s not possi bl e to T
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Identification of Constituents of Fennel Teas J. Agric. Food Chem., Vol. 48, No. 10, 2000 4737
obtai n such an essenti al oi l content i n the i nfusi ons, our
resul ts l ead to the concl usi on that none of the tested
teas contai n such essenti al oi l i n an amount suffi ci ent
to expl ai n the secretol yti c and expectorant purposes.
However, because the putati ve pharmacol ogi cal prop-
erti es of the i nfusi ons have not yet been compl etel y
veri fi ed, experi mental evi dence of the pharmacol ogi cal
properti es are needed for an apprai sal of a possi bl e effect
of other consti tuents of the teas such as fl avonoi ds and
chl orogeni c aci d.
LI TERATURE CI TED
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tropi c acti on of i nhal ed terpenes and rel ated vol ati l e agents.
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Vol . 5, pp 156-181.
Brol i s, M.; Gabetta, B.; Fuzzati , N.; Pace, R.; Panzeri , F.;
Peterl ongo, F. I denti fi cati on by hi gh-performance l i qui d
chromatography-di ode array detecti on-mass spectrometry
and quanti fi cati on by hi gh performance l i qui d chromatog-
raphy-UV absorbance detecti on of acti ve consti tuents of
Hypericumperforatum. J . Chromatogr., A 1998, 825, 9-16.
Czygan, F. C. Fenchel . I n Teedrogen, 2nd ed.; Wi chtl , M., Ed.;
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171-173.
Dorsch, W.; Loew, D.; Meyer, E.; Schi l cher, H. I n Empfehlun-
gen zu Kinderdosierungen von monographierten Arzneidro-
gen und ihren Zubereiungen; Kooperati on Phytopharmaca:
Bonn, 1993; pp 50-51.
Forster, H. B. Spasmol yti sche Wi rkung pfl anzl i cher Carmi -
nati va. Z. All. Med. 1983, 59, 1327-1333.
Forster, H. B.; Ni kl as, H.; Lutz, S. Anti spasmodi c effects of
some medi ci nal pl ants. Planta Med. 1980, 40, 309-319.
Kunzemann, J.; Herrmann, K. I sol ati on and i denti fi cati on of
fl avon(ol )-O-gl ycosi des i n caraway (Carumcarvi L.), fennel
(FoeniculumvulgareM.), ani se (Pimpinella anisumL.), and
cori ander (Coriandrum sativum L.) and of fl avonol -C-
gl ycosi des i n ani se. I Phenol i cs of spi ces. Z. Lebensm.-
Unters.-Forsch. 1977, 164, 194-200.
Madaus, G. Foeni cul um. I n Lehrbuch der biologischen Heil-
mittel, Vol . 2; G. Ol ms, Ed.; Hi l eshei m: New York, 1976;
pp 1354-61.
Merkes, K. Drogen mi t atheri schem O l (XVI ) Foeni cul um
vul gare Mi l l er- Fenchel . PTA-Repetitorium 1980, 12, 45-
48.
Murray, R. D. H.; Mendez, J.; and Brown, S. A. I n TheNatural
Coumarins. Occurrence, Chemistry, Biochemistry Wi l ey-
I ntersci ence, John Wi l ey & Sons Ltd.: Chi chester, 1982; p
560.
Schuster, K. P. Di ssertati on, Uni versi ty of Muni ch, Muni ch,
Germany, 1971.
Toth, L. Untersuchungen uber das atheri sche O l von Foen-
iculumvulgare. Planta Med. 1967, 15, 371-89.
Trenkl e, K. Recent studi es on fennel (FoeniculumvulgareM.)
2. The vol ati l e oi l of the frui t, herbs and roots of frui t-beari ng
pl ants. Pharmazie1972, 27, 319-24.
Reynol ds, J. E. F. Fennel , fennel oi l . I n Martinadale-TheExtra
Pharmacopoeia, 30th ed. The Pharmaceuti cal Press: Lon-
don, 1993; pp 1369-1370.
Wei b, R. F. I n Lehrbuch der Phytoterapie, 7th ed. Hi p-
pokrates: Stuttgart, 1991; pp 107-108.
Recei ved for revi ew February 10, 2000. Revi sed manuscri pt
recei ved June 15, 2000. Accepted June 16, 2000. Thi s work
was supported by M.U.R.S.T. (Mi ni stero Uni versi ta` Ri cerca
Sci enti fi ca e Tecnol ogi ca, Roma).
JF000171N
4738 J. Agric. Food Chem., Vol. 48, No. 10, 2000 Bilia et al.