Amines in the Marking Fluid and Anal Sac Secretion of the Tiger, Panthera tigris G lyn R. Banks, A lan J. Buglass, and Jo h n S. W aterhouse Anglia Polytechnic University, East Rd., Cambridge, CB 1 1 PT, U.K. Z. Naturforsch. 47c, 618-620 (1992); received March 12, 1992 Amines, Anal Sac Secretion, Marking Fluid, Scent-Marking, Tiger Analysis of the marking fluid of two tigers (one Bengal and one Sumatran) by GC using an amine-specific column and a nitrogen-specific detector has shown the presence of the follow ing amines: ammonia, methylamine, dimethylamine, trimethylamine, triethylamine, propyl amine, and butane-1,4-diamine (putrescine). In contrast to previously published reports, we were unable to detect 2-phenylethylamine. The anal sac secretion was found to have a similar amine content.
Introduction Tigers use a variety o f scent-m arkings in the wild. Sm ith et al. [1], on the basis o f their field studies o f tigers in N epal, list several m odes o f scent-m arking: scrapes, scats, anal sac secretion, urine stains on trees and shrubs, claw -raking and cheek-rubbing. T he m ajor purpose o f these m ark ings seems to be for the establishm ent an d m ainte nance o f territo ry [1]. A n o th er form o f scentm arking, n o t m entioned explicitly in the above re p o rt, has been described by Schaller [2] and by B rahm achary an d D u tta [3 -5 ], and is considered by the latter au th o rs to be semiochem ically the m ost significant m ethod o f m arking. This com prises a w hite fatty fluid, term ed ‘m arking fluid’ [3 -5 ], w hich is sprayed at an angle as a jet, inde p endent o f urine, from the u rinary tract o f b oth m ale an d fem ale tigers w hen the hind leg is raised [4], This action deposits the m arking fluid on vege ta tio n 1- 1 .3 m above the ground. As p ointed o u t [5], there has been som e confu sion in the literature betw een anal sac secretion an d m arking fluid, even tho u g h they are distinctly different in ap p earance and origin. D eposits o f anal sac secretion are described as being d a rk and w axy w ith a distinctive o d o u r [ 1 ], w hereas m arking fluid is w hite an d m uch m ore m obile w ith an amm oniacal o dour. A nal sac secretion is deposited w ith faeces; an d as there is no connection between the anal gland and the urinary tract, there is no Reprint requests to Dr. J. S. Waterhouse. Verlag der Zeitschrift für Naturforschung, D-W-7400 Tübingen 0939-5075/92/0700-0618 $01.30/0
w ay in w hich an al gland secretion can be em itted w ith urine. U rine, on the o th er hand, m ay contain sm all am o u n ts o f m arking fluid [4], A nalysis o f steam distillate extracts o f m arking fluid from b o th m ale and fem ale tigers using TLC an d p aper chro m ato g rap h y has been carried out [3, 4], The chro m ato g ram s revealed the presence o f several am ines, o f w hich 2 -phenylethylam ine was rep o rted to be the m ajor com ponent in each ani m al. O ther am ines identified were butane-1,4-di am ine (putrescine) and pentane-1,5-diam ine (cadaverine) as well as am m onia [5]. N o analytical w ork on the an al sac secretion o f the tiger has yet been published. As p art o f o u r long-term interest in the com posi tion o f sem iochem ically im p o rtan t glandular se cretion o f m am m als, we have analyzed b oth the m arking fluid and anal sac secretion o f the tiger specifically for the presence o f am ines, and we present here o u r prelim inary findings. Materials and Methods M arking fluid from tw o female tigers, K iaw ang (Bengal) and Suzy (S um atran), were collected from their enclosures in L ondon Zoo, R egent’s P ark. A nal sac secretion from tiger K iaw ang was o b tained directly from the anal sac while the ani m al was anaesthetized. All sam ples were stored at - 1 5 °C until analyzed. A nalyses were carried out on a V arian 3700 gas ch ro m ato g rap h equipped w ith a glass colum n (2 m long, 2 m m ID ) packed w ith 60/80 C arb o p ack B /4% C arbow ax 20 m / 0.8% K O H (Supelco). The detector was a th e r m ionic d etector for the specific detection o f n itro gen. Sam ples ( ~ 100 mg) o f m arking fluid and anal
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G. R. Banks et al. ■Amines in Tigers
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sac secretion were dissolved in 0.05 m N a O H so lu tion (1.0 ml), an d 4 jil injected o n to the colum n. The colum n tem p eratu re was held at 50 °C for 2 min, an d was then raised to 210 °C a t 50 °C /m in. The nitrogen flow rate w as 30 m l/m in.
Results and Discussion The gas ch ro m ato g ram from a sam ple o f m a rk ing fluid from K iaw ang (collected 7 .1 1 .8 9 ) is show n in Fig. 1. The identities o f the am ines were determ ined by com p ariso n o f retention tim es w ith those o f authentic sam ples. T he m ajo r am ine in the secretion is trim ethylam ine, w ith a significant am o u n t o f butane-1,4-diam ine an d sm aller am ounts o f m ethylam ine, dim ethylam ine a n d triethylam ine. In stark co n tra st to previously p u b lished w ork [3, 4], the c h ro m ato g ram show s only a possible h in t o f 2 -phenylethylam ine at a reten tio n time o f 12 m in. T o check th a t we h ad n o t m issed this am ine, 2-phenylethylam ine (0.5 mg) w as a d d ed to a sam ple o f m ark in g fluid ( 1 0 0 m g) a n d the m ixture analyzed u n d er the sam e conditions. T he resulting ch ro m ato g ram is show n in Fig. 2, in which the large peak due to 2 -phenylethylam ine has a reten tio n tim e o f 12 m in. This result clearly shows th a t 2 -phenylethylam ine is n o t p resen t in this sam ple o f secretion to any significant extent. A second sam ple o f secretion tak en from th e sam e tiger a t a later d ate (21.2.91) revealed th e sam e suite o f am ines, the m ain difference betw een the
Elution time [ min ]
Fig. 1. Gas chromatogram of marking fluid from tiger Kiawang collected 7.11.89. Compounds identied as fol lows: 1, ammonia; 2, methylamine; 3, dimethylamine; 4, trimethylamine; 5, propylamine; 6, triethylamine; 7, butane-1,4-diamine.
Elution time [ min ]
Fig. 2. Gas chromatogram of marking fluid from tiger Kiawang collected 7.11.89 with added 2-phenylethyl amine (compound 8). Compounds 1 - 7 as in Fig. 1.
two sam ples being the reduced am o u n t o f b utane1,4-diamine in the latter (Fig. 3). T he reason for this difference is n ot certain, b ut it does indicate th a t the com position o f secretion from an individ ual does vary. The chro m ato g ram o b tained from the m arking fluid o f tiger Suzy is show n in Fig. 4. This is alm ost identical to the ch ro m ato g ram o f the first sam ple taken from K iaw ang, despite the fact th a t the anim als were from different sub species, and again, there is no indication o f 2 -phenylethylam ine. Fig. 5 shows the ch ro m ato g ram o btained from the anal sac secretion o f K iaw ang. A lth o u g h this
Elution time [ min ]
Fig. 3. Gas chromatograph of marking fluid from tiger Kiawang collected 21.2.91. Compounds 1 - 7 as in Fig. 1.
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G. R. Banks et al. ■Amines in Tigers
Elution time [ min ]
Fig. 4. Gas chromatogram of marking fluid from tiger Suzy. Compounds 1 - 7 as in Fig. 1.
d ark, waxy m aterial is so different in its ap p ear ance from the w hite, m obile m arking fluid, it has an am ine profile th a t is strikingly sim ilar to th a t o f the latter. O ne o f the few differences is the pres ence o f a significant am o u n t o f iso-butylam ine in the anal sac secretion. A gain, 2-phenylethylam ine was n ot observed. The great sim ilarity in the am ine contents o f the m arking fluid an d anal sac secretion was unexpect ed and surprising, an d it raises the question o f the relationship betw een these tw o excretions, seem ingly so different in b o th appearance and m ode o f deposition. D o they carry the sam e sem iochem ical message? It is also difficult to u n d erstan d the great discrepancy betw een our results and previously published rep o rts [3 -5 ] concerning the presence o f 2-phenylethylam ine in the m arking fluid. A l though o u r w o rk and previous studies [4] show th a t some v ariatio n is to be expected betw een indi viduals an d over tim e for the sam e individual, the dichotom y regarding 2 -phenylethylam ine w ould seem to be to o great to be explained in these term s. It m ay be th a t the absence o f 2-phenylethylam ine in o u r tigers reflects a m etabolic difference be tween them a n d the tigers previously studied in India [3 -5 ].
A cknowledgemen ts Elution time [ min ]
Fig. 5. Gas chromatograph of the anal sac secretion of tiger Kiawang. Compounds 1 -7 as in Fig. 1; 9, iso-butylamine; 10, pentane-1,5-diamine; 11, butylamine.
[1] J. L. D. Smith, C. McDougal, and D. Miquelle, Anim. Behav. 37, 1 (1989). [2] G. B. Schaller, The deer and the tiger, Univ. Chicago Press, Chicago 1976. [3] R. L. Brahmachary and J. Dutta, Z. Naturforsch. 3 4 c , 632(1979).
W e are grateful to D r. C hris F aulkes o f L o ndon Z oo for supply o f m arking fluid and anal sac secre tion.
[4] R. L. Brahmachary and J. Dutta, Am. Nat. 118, 561 (1981). [5] R. L. Brahmachary and J. Dutta, in: Tigers o f the world (R. L. Tilson and U. S. Seal, eds.), pp. 2 9 6 302, Noyes Press, New Jersey 1987.
Unauthenticated Download Date | 1/18/19 4:05 AM
Introduction Tigers use a variety o f scent-m arkings in the wild. Sm ith et al. [1], on the basis o f their field studies o f tigers in N epal, list several m odes o f scent-m arking: scrapes, scats, anal sac secretion, urine stains on trees and shrubs, claw -raking and cheek-rubbing. T he m ajor purpose o f these m ark ings seems to be for the establishm ent an d m ainte nance o f territo ry [1]. A n o th er form o f scentm arking, n o t m entioned explicitly in the above re p o rt, has been described by Schaller [2] and by B rahm achary an d D u tta [3 -5 ], and is considered by the latter au th o rs to be semiochem ically the m ost significant m ethod o f m arking. This com prises a w hite fatty fluid, term ed ‘m arking fluid’ [3 -5 ], w hich is sprayed at an angle as a jet, inde p endent o f urine, from the u rinary tract o f b oth m ale an d fem ale tigers w hen the hind leg is raised [4], This action deposits the m arking fluid on vege ta tio n 1- 1 .3 m above the ground. As p ointed o u t [5], there has been som e confu sion in the literature betw een anal sac secretion an d m arking fluid, even tho u g h they are distinctly different in ap p earance and origin. D eposits o f anal sac secretion are described as being d a rk and w axy w ith a distinctive o d o u r [ 1 ], w hereas m arking fluid is w hite an d m uch m ore m obile w ith an amm oniacal o dour. A nal sac secretion is deposited w ith faeces; an d as there is no connection between the anal gland and the urinary tract, there is no Reprint requests to Dr. J. S. Waterhouse. Verlag der Zeitschrift für Naturforschung, D-W-7400 Tübingen 0939-5075/92/0700-0618 $01.30/0
w ay in w hich an al gland secretion can be em itted w ith urine. U rine, on the o th er hand, m ay contain sm all am o u n ts o f m arking fluid [4], A nalysis o f steam distillate extracts o f m arking fluid from b o th m ale and fem ale tigers using TLC an d p aper chro m ato g rap h y has been carried out [3, 4], The chro m ato g ram s revealed the presence o f several am ines, o f w hich 2 -phenylethylam ine was rep o rted to be the m ajor com ponent in each ani m al. O ther am ines identified were butane-1,4-di am ine (putrescine) and pentane-1,5-diam ine (cadaverine) as well as am m onia [5]. N o analytical w ork on the an al sac secretion o f the tiger has yet been published. As p art o f o u r long-term interest in the com posi tion o f sem iochem ically im p o rtan t glandular se cretion o f m am m als, we have analyzed b oth the m arking fluid and anal sac secretion o f the tiger specifically for the presence o f am ines, and we present here o u r prelim inary findings. Materials and Methods M arking fluid from tw o female tigers, K iaw ang (Bengal) and Suzy (S um atran), were collected from their enclosures in L ondon Zoo, R egent’s P ark. A nal sac secretion from tiger K iaw ang was o b tained directly from the anal sac while the ani m al was anaesthetized. All sam ples were stored at - 1 5 °C until analyzed. A nalyses were carried out on a V arian 3700 gas ch ro m ato g rap h equipped w ith a glass colum n (2 m long, 2 m m ID ) packed w ith 60/80 C arb o p ack B /4% C arbow ax 20 m / 0.8% K O H (Supelco). The detector was a th e r m ionic d etector for the specific detection o f n itro gen. Sam ples ( ~ 100 mg) o f m arking fluid and anal
Unauthenticated Download Date | 1/18/19 4:05 AM
G. R. Banks et al. ■Amines in Tigers
619
sac secretion were dissolved in 0.05 m N a O H so lu tion (1.0 ml), an d 4 jil injected o n to the colum n. The colum n tem p eratu re was held at 50 °C for 2 min, an d was then raised to 210 °C a t 50 °C /m in. The nitrogen flow rate w as 30 m l/m in.
Results and Discussion The gas ch ro m ato g ram from a sam ple o f m a rk ing fluid from K iaw ang (collected 7 .1 1 .8 9 ) is show n in Fig. 1. The identities o f the am ines were determ ined by com p ariso n o f retention tim es w ith those o f authentic sam ples. T he m ajo r am ine in the secretion is trim ethylam ine, w ith a significant am o u n t o f butane-1,4-diam ine an d sm aller am ounts o f m ethylam ine, dim ethylam ine a n d triethylam ine. In stark co n tra st to previously p u b lished w ork [3, 4], the c h ro m ato g ram show s only a possible h in t o f 2 -phenylethylam ine at a reten tio n time o f 12 m in. T o check th a t we h ad n o t m issed this am ine, 2-phenylethylam ine (0.5 mg) w as a d d ed to a sam ple o f m ark in g fluid ( 1 0 0 m g) a n d the m ixture analyzed u n d er the sam e conditions. T he resulting ch ro m ato g ram is show n in Fig. 2, in which the large peak due to 2 -phenylethylam ine has a reten tio n tim e o f 12 m in. This result clearly shows th a t 2 -phenylethylam ine is n o t p resen t in this sam ple o f secretion to any significant extent. A second sam ple o f secretion tak en from th e sam e tiger a t a later d ate (21.2.91) revealed th e sam e suite o f am ines, the m ain difference betw een the
Elution time [ min ]
Fig. 1. Gas chromatogram of marking fluid from tiger Kiawang collected 7.11.89. Compounds identied as fol lows: 1, ammonia; 2, methylamine; 3, dimethylamine; 4, trimethylamine; 5, propylamine; 6, triethylamine; 7, butane-1,4-diamine.
Elution time [ min ]
Fig. 2. Gas chromatogram of marking fluid from tiger Kiawang collected 7.11.89 with added 2-phenylethyl amine (compound 8). Compounds 1 - 7 as in Fig. 1.
two sam ples being the reduced am o u n t o f b utane1,4-diamine in the latter (Fig. 3). T he reason for this difference is n ot certain, b ut it does indicate th a t the com position o f secretion from an individ ual does vary. The chro m ato g ram o b tained from the m arking fluid o f tiger Suzy is show n in Fig. 4. This is alm ost identical to the ch ro m ato g ram o f the first sam ple taken from K iaw ang, despite the fact th a t the anim als were from different sub species, and again, there is no indication o f 2 -phenylethylam ine. Fig. 5 shows the ch ro m ato g ram o btained from the anal sac secretion o f K iaw ang. A lth o u g h this
Elution time [ min ]
Fig. 3. Gas chromatograph of marking fluid from tiger Kiawang collected 21.2.91. Compounds 1 - 7 as in Fig. 1.
Unauthenticated Download Date | 1/18/19 4:05 AM
620
G. R. Banks et al. ■Amines in Tigers
Elution time [ min ]
Fig. 4. Gas chromatogram of marking fluid from tiger Suzy. Compounds 1 - 7 as in Fig. 1.
d ark, waxy m aterial is so different in its ap p ear ance from the w hite, m obile m arking fluid, it has an am ine profile th a t is strikingly sim ilar to th a t o f the latter. O ne o f the few differences is the pres ence o f a significant am o u n t o f iso-butylam ine in the anal sac secretion. A gain, 2-phenylethylam ine was n ot observed. The great sim ilarity in the am ine contents o f the m arking fluid an d anal sac secretion was unexpect ed and surprising, an d it raises the question o f the relationship betw een these tw o excretions, seem ingly so different in b o th appearance and m ode o f deposition. D o they carry the sam e sem iochem ical message? It is also difficult to u n d erstan d the great discrepancy betw een our results and previously published rep o rts [3 -5 ] concerning the presence o f 2-phenylethylam ine in the m arking fluid. A l though o u r w o rk and previous studies [4] show th a t some v ariatio n is to be expected betw een indi viduals an d over tim e for the sam e individual, the dichotom y regarding 2 -phenylethylam ine w ould seem to be to o great to be explained in these term s. It m ay be th a t the absence o f 2-phenylethylam ine in o u r tigers reflects a m etabolic difference be tween them a n d the tigers previously studied in India [3 -5 ].
A cknowledgemen ts Elution time [ min ]
Fig. 5. Gas chromatograph of the anal sac secretion of tiger Kiawang. Compounds 1 -7 as in Fig. 1; 9, iso-butylamine; 10, pentane-1,5-diamine; 11, butylamine.
[1] J. L. D. Smith, C. McDougal, and D. Miquelle, Anim. Behav. 37, 1 (1989). [2] G. B. Schaller, The deer and the tiger, Univ. Chicago Press, Chicago 1976. [3] R. L. Brahmachary and J. Dutta, Z. Naturforsch. 3 4 c , 632(1979).
W e are grateful to D r. C hris F aulkes o f L o ndon Z oo for supply o f m arking fluid and anal sac secre tion.
[4] R. L. Brahmachary and J. Dutta, Am. Nat. 118, 561 (1981). [5] R. L. Brahmachary and J. Dutta, in: Tigers o f the world (R. L. Tilson and U. S. Seal, eds.), pp. 2 9 6 302, Noyes Press, New Jersey 1987.
Unauthenticated Download Date | 1/18/19 4:05 AM
