Supplementary Information.
New Mantellid Alkaloids, previously undescribed:
151C. Unclass. C10H17N. Rt 4.70. MS: 151(55), 150(26), 136(100), 134(18), 122(27), 94(28), 80(16). Table 5.
197J. 3,5-P. ‘C12H23NO’. Rt 8.82. MS: 197(17), 182(7), 124(100), 110(15), 96(9). Table 3
199B. Unclass. ‘C12H25NO’. Rt 7.64. MS: 199(23), 198(18), 184(100), 113(29), 112(24), 111(21), 72(23), 58(55) Table 3.
205L. Dehydro-5,8-I. ‘C14H23N’. Rt 7.45. MS: 205(1), 136(100), 134(70), 120(6). Table 1.
207W. Dehydro-5,8-I. ‘C14H25N’. Rt 9.67. MS: 207(2), 206(13), 136(100), 134(63), 120(11). Table 4.
217H, Tricyclic. ‘C15H23N’. Rt 9.32. MS: 217(48), 202(16), 188(87), 178(30), 160(33), 120(55), 70(100), 0D.
231N. Tricyclic. ‘C16H25N’. Rt 10.73. MS: 231(21), 230(55), 216(10), 202(100), 191(68), 190(87), 174(70), 160(61), 134(81), 122(87), 110(39), 84(88). Table 5.
237V. Unclass. ‘C15H27NO’. Rt 12.82. MS: 237(10), 236(31), 222(8), 136(100), 122(54), 70(17). Table 5.
239Z. 5,6,8-I. ‘C15H29NO’. Rt 11.62. MS: 239(21), 238(1), 196(100), 124(12), 70(3), 1D. Table 4.
239BB. 3,5-P. ‘C15H29NO’. Rt 13.36 > 12.58. MS: 239(2),124(100). Perhaps isomers of 239K with terminal CH2OH side-chain or N-oxides. Table 6.
247N. Tricyclic. ‘C17H29N’. Rt 11.73. MS: 247(50), 246(68), 232(33), 218(100), 204(28), 190(32), 176(57), 150(23), 148(48), 120(20), 96(14), 80(15), 70(16). Table 3.
247O. Dehydro-5,8-I. ‘C15H29N’. Rt 12.00. MS: 247(6), 245(5), 218(12), 150(100), 148 (38), 120(9), 0D. Tables 5, 6.
249AA. Unclass. ‘C16H25NO’. Rt 11.28. MS: 249(12), 204(16), 178(100), 148(7), 128(9), 96(9), 70(8), 1D. Table 2.
249BB. 5,6,8-I. ‘C17H31N’. Rt 10.12. MS: 249(6), 234(2), 152(100), 110(8), 70(5). 0D. Table 2.
263S. 3,5-P. ‘C18H33N’. Rt 12.40. MS: 12.40. MS: 263(8), 220(6), 124(100). Table 3.
263T. Tricyclic. ‘C17H29NO’. Rt 14.05. MS: 263(32), 262(48), 248(56), 234(88), 190(25), 176(63), 154(100), 150(50), 148(45), 124(43), 110(30), 70(20). Table 3.
265CC. Tricyclic. ‘C15H31NO’. Rt 14.28. MS: 265(22), 264(48), 224(73), 222(33), 180(26), 166(23), 152(100), 138(35), 124(43), 110(56), 98(61), 96(60). Table 5.
269J. 5,6,8-I. ‘C16H31NO2’. Rt 14.76. MS: 269(2), 268(11), 212(100), 194(4), 140(6), 70(3). Table 4.
279L. DeoxyPTX. ‘C17H29NO2’. Rt 13.01. MS: 279(7), 278(24), 264(11), 178(14), 150(100), 70(9). Table 7.
289G. 5,6,8-I. ‘C20H35N’. Rt 14.79. MS: 289(2), 288(1), 234(21), 222(8), 152(100), 110(14), 70(4). Table 3.
293O. Dehydro-5,8-I. ‘C19H35NO’. Rt 15.43. MS: 293(7), 250(4), 236(6), 178(100), 176(27), 120(7), Table 3.
297F. Unclass. ‘C17H31NO3’ Rt 14.67. MS: 297(2), 280(2), 238(7), 212(100), 194(37), 128(42), 70(30). 3D. Table 5.
307K. DeoxyPTX. ‘C19H33NO2’. Rt 15.34. MS: 307(<1), 306(3), 192(7), 164(100), 84(7). Table 5.
307L. Unclass. ‘C19H33NO2’. Rt 14.63. MS: 307(36), 306(14), 250(23), 210(34), 180(100), 178(33), 136(15), 122(23), 108(15), 70(56). Table 2.
307M. Unclass. ‘C19H33NO2’. Rt 16.67. MS: 307(3), 290(100), 264(46), 250(43), 237(27), 122(25), 110(30), 70(46). Table 7.
309K. Dehydroizidine. ‘C19H33NO2’. Rt 17.18. MS: 309(5), 308(3), 294(3), 250(26), 194(100), 190(65), 136(16), 134(19), 70(2). Table 3, 6.
323J. aPTX. ‘C19H33NO3’. Rt 16.75. MS: 323(4), 306(21), 288(12), 207(15), 182(100), 114(26), 112(14), 70(94). 2D. Table 2.
Comments on the data of Clark, et al., 2006:
Total ion current (TIC) chromatograms of all 24 extracts analyzed by Clark, et al., 2006were reproduced in their Supplementary Information with identification of the base peak in EI-MS and protonated parent ion in CI-MS. The data, part of which appears in captions to the chromatograms and more of which appears in Table 1 of that paper are indicated as major/minor/trace alkaloids. The Table 1 data, particularly, often appears at variance with the TIC chromatograms. Many alkaloid peaks are ignored (even for known alkaloids), some are misassigned and often the levels reported in Table 1 do not correspond to the peak intensities in the chromatograms. Some chromatograms are truncated so that alkaloid peaks with short or more often long retention times cannot be assessed.
Further complicating the proposal of Clark et al. (2006) that there would be less alkaloids in disturbed sites, was the inclusion of at least four individuals of M. baroni from a disturbed site, in which alkaloids were obtained solely by electrical stimulation (Transcutaneous Amphibian Stimulation, TAS). Although the authors claim that the total number of alkaloids extracted from an individual are the same using the TAS or the standard extraction methods, the TAS method appears to yield extracts with lesser amounts of alkaloids (see Supplementary Information, Clark et al., 2006 to compare total ion chromatograms of extracts obtained by the TAS procedure with those obtained by extraction of skin). Thus, the inclusion of the four TASed samples, all from disturbed sites and all with low numbers of detected alkaloids, results in what we consider an erroneous estimate of lower alkaloid numbers for disturbed sites.
GPS coordinates forour collection sites:
Table
/Site/ (date)
/Elevation (m)
/Latitude
/Longitude
/Condition
1 / Ranomafana(Ranomafanakely) / 1134 / 21.25S. / 47.37E. / Pristine/ slightly disturbed
3 / “ / ” / “ / “ / Disturbed
2 / Besariaka / 19.13S. / 48.28E. / Disturbed
4 / “ / ” / “ / “ / “
5 / Manombo / 44 / 23.03S. / 47.73E. / Disturbed
6 / 581 / 22.79S. / 47.19E. / Disturbed
7 / Ranomafana / 1019 / 21.26S. / 47.40E.
7 / (Feb. 21, ‘03) / 18.61S. / 48.33E.
7 / Close to Tsinjarivo (Feb. 19-20, ’03) / 19.63S. / 47.68E. / Very Disturbed
7 / Vohindrazana (Feb. 19-20, ’03) / 810 / 18.97S. / 48.51E.
7 / Vohiparara (Jan. 16, ’03); / 1152 / 21.23S. / 47.37E. / Pristine
8 / Ranomafana (Jan. 24, ’03) / 21.39S. / 47.46E. / Pristine
Voucher numbersforrepresentativecollectionsa:
Table 1 / Table 3 / Table 5 / Table 6voucher #
ZMA / ZCMV code / voucher #
ZMA / ZCMV code / voucher #
ZMA / ZCMV code / voucher #
ZMA / ZCMV code
UADBA-uncat. / 126 / 20289 / 171 / 20151 / 502 / 20175 / 701
20275 / 127 / 20290 / 172 / 20152 / 503 / 20189 / 702
20276 / 128 / 20326 / 173 / 20153 / 504 / 20176 / 703
20277 / 129 / 20327 / 174 / 20154 / 505 / 20177 / 704
20278 / 130 / 20328 / 175 / 20155 / 506 / 20178 / 705
20279 / 131 / 20329 / 176 / 20156 / 507 / 20179 / 706
20280 / 132 / UADBA-uncat. / 501 / 20180 / 707
20281 / 133 / 20157 / 508 / 20181 / 708
20282 / 134 / 20158 / 509 / 20182 / 709
20283 / 135 / 20159 / 510 / 20183 / 710
20284 / 136 / 20160 / 520 / 20184 / 711
20285 / 137 / 20161 / 521 / 20185 / 712
20286 / 138 / 20162 / 522 / 20186 / 713
20287 / 139 / 20163 / 523 / 20187 / 714
20288 / 140 / 20164 / 524 / 20188 / 715
20173 / 620 / 20190 / 901
20174 / 526 / 20261 / 902
20167 / 525 / 20262 / 903
20263* / 527 / 20268 / 904
20174* / 621 / 20269 / 905
20164 / 529 / 20271 / 906
20165 / 528 / 20272 / 907
20168 / 530 / 20273 / 909
20169 / 531 / 20274 / 910
20170 / 532
UADBA-uncat. / 413
aNo vouchers exist for Tables 2, 4, 8,and 9. Abbreviations: UADBA, Université d’Antananarivo, Dept. de Biologie Animale; ZMA, Zoological Museum of Amsterdam; ZCMV, Zoological Collection of Miguel Vences (field numbers). *Specimens may have been interchanged.