Identification and differentiation of dragon s blood in works of art using gas chromatography/mass spectrometry

Analytical and Bioanalytical Chemistry Electronic Supplementary Material Identification and differentiation of dragon s blood in works of art using gas chromatography/mass spectrometry Ursula Baumer and Patrick Dietemann 1/10

Table S1: Provenance of the dragon s blood reference materials analysed in this study. Materials from botanically identified plants are marked in grey, the others are attributed according to the analyses. Reference collection Dating Identified species Collection Theodor Martius, Friedrich Alexanders-University Erlangen (all samples inscribed resina sanguinis draconis) Royal Botanic Garden Kew, Economic Botany Collection Kew Botanisches Museum Berlin- Dahlem, Harzsammlung Botanischer Garten München, Herbarium Dragons blood from Malaysia, Forest Research Institute Malaysia (FRIM), Kepong, Selangor Darul Ehsan, Malaysia Dragon s blood Gran Canaria (Katja Lewerentz, Cologne University of Applied Sciences) Dragon s blood Socotra, Stuttgart State Academy of Art and Design Dragon s blood from Socotra (Katja Lewerentz, Cologne University of Applied Sciences) Dragon s blood from Sumatra, Dr. Psota, Bern, Switzerland Apotheke Spaichingen (historic pharmacy collection, Cologne University of Applied Sciences) 1825-1863 Daemonorops draco (inv.-no. 40.161, 40.164, 40.166, 40.168, 40.169, 40.171, 40.207) Daemonrops micracantha (inv.-no. 40.159) Dracaena cinnabari (inv.-no. 40.160, 40.162) Pterocarpus santalinus (inv.-no. 27.1.3) Croton (inscribed Resina sanguinis draconis westindisch, inv.-no. 40.163, 40.167) 1896 Daemonorops draco (inscribed Dae. propinqua Becc. a, EBC 35500) 1932 Daemonrops micracantha (inscribed Daemonorops Draco Blume, EBC 35499) b Daemonorops draco (inscribed Dae. Propinquus a, EBC 35519) b Dracaena cinnabari (EBC 61201, EBC 61195) 1957 Pterocarpus Santalinus L. f. (EBC 21364) b Pterocarpus Indicus Willd. (EBC 21359) b Croton sp. Euphorbiaceae (EBC 43900) 20 th century Dracaena cinnabari Balf. fil. (inv.-no. B_16_000010, B_16_000001, B_16_000011, B_16_000007) Dracaena draco (L.) L. (inv.-no. B_16_000005) Daemonorops draco (inv.-no. B_16_000009, B_16_000008, B_16_000013, B_16_000012) 1930 Dracaena draco (L.) L. 1999 Dracaena draco L. subsp. ajgal 2000 Daemonorops micracantha 2000 Dracaena draco 2006, 1994 Dracaena cinnabari 1998 Dracaena cinnabari 2003 Daemonorops draco ~ 1900 Daemonorops draco 2/10

Dragon s blood India/Sumatra, Kremer Pigmente, Aichstetten, Germany Dragon s blood cake, from Socotra, Kremer Pigmente, Aichstetten Dragon s blood powder, GED Gerhard Eggebrecht, Süderau, Germany Dragon s blood ball, GED Gerhard Eggebrecht Sandalwood, GED Gerhard Eggebrecht, Art. No. 24-860 Fa. Bluegreen, Vienna, Art. No. 244 ~ 2000 Daemonorops draco (prod.-no. 3700) 1994 Shellac with Dracaena cinnabari (prod.- no. 37011) ~ 2000 Daemonorops draco (prod.-no. D22-270) ~ 2000 Daemonorops draco (prod.-no. D22-260) 2000 Pterocarpus santalinus 2000 Croton a Daemonorops propinquus has recently been reported to be synonymous to Daemonorops draco [26]. b donation date not recorded Table S2: Historic commercial packing of dragon s blood in the Martius collection and its botanic origin. Samples date around 1825 to 1863. Latin names of historic commercial packing in the Martius collection Resina sanguinis draconis in granis (grains) Resina sanguinis draconis in lacrimis (tears) Resina sanguinis draconis in baculis (bars) Resina sanguinis draconis in globuli (globes) Resina sanguinis draconis in massis (blocks) Approximate dimensions Botanical origin identified after analysis 1-2 cm Dracaena cinnabari 2-4 cm Dracaena cinnabari 1-2 cm, 20-30 cm length 20-30 cm 2-3 cm height x 15-30 cm length Daemonorops draco Sandalwood Daemonorops draco 3/10

Table S3-a: Mass spectral data of compounds identified in dragon's blood of Dracaena cinnabari. No. 10 most abundant MS peaks MW tentative assignment Ref. DrC1 226 (100), 104 (45), 225 (24), 209 (18), 135 (17), 91 (17), 227 (17), 211 (13), 117 (13), 115 (10) DrC2 240 (100), 107 (83), 79 (65), 133 (59), 258 (52), 151 (35), 209 (28), 126 (25), 138 (22), 91 (22) DrC3 137 (100), 256 (51), 124 (34), 105 (26), 77 (23), 107 /19), 151 (11), 138 (10), 257 (9), 240 (5) DrC4 240 (100), 136 (77), 163 (71), 239 (71), 104 (42), 108 (30), 103 (18), 137 (15), 241 (15), 78 (14) DrC5 256 (100), 152 (92), 117 (45), 240 (33), 239 (32), 124 (28), 104 (28), 91 (26), 136 (24), 69 (23) DrC6 256 (100), 179 (83), 152 (64), 255 (62), 124 (36), 137 (33), 107 (20), 153 (18), 257 (16), 258 (16) DrC7 242 (100), 120 (80), 123 (55), 107 (28), 136 (26), 133 (17), 243 (17), 91 (14), 119 (14), 121 (13) DrC8 150 (100), 272 (79), 137 (35), 135 (34), 151 (19), 123 (18), 136 (18), 122 (14), 273 (13), 107 (13) DrC9 256 (100), 131 (39), 107 (30), 132 (27), 137 (21), 225 (19), 133 (17), 257 (17), 255 (15), 239 (14) DrC10 256 (100), 107 (97), 148 (64), 147 (42), 149 (36), 108 (22), 123 (22), 77 (20), 133 (18), 257 (17) DrC11 286 (100), 107 (73), 178 (37), 147 (26), 153 (24), 150 (22), 179 (21), 272 (21), 287 (19), 177 (15) DrC12 286 (100), 150 (69), 135 (37), 137 (37), 151 (34), 178 (19; 287 (18), 136 (16), 162 (16), 163 (16) DrC13 138 (100), 286 (52), 137 (51), 123 (22), 106 (17), 147 (15), 139 (10), 148 (10), 287 (9), 149 (9) DrC14 284 (100), 107 (68), 176 (47), 151 (30), 177 (24), 147 (24), 175 (21), 133 (19), 150 (18), 285 (17) DrC15 138 (100), 286 (49), 137 (33), 123 (19), 147 (15), 106 (14), 139 (11), 316 (11), 287 (9), 107 (8) DrC16 316 (100), 150 (82), 167 (78), 137 (35), 135 (23), 151 (19), 164 (18), 317 (18), 138 (17), 165 (16) 226 7-hydroxyflavan [42] 258 256 4-hydroxy-2-methoxy-dihydrochalcone [42] 240 7-hydroxyflavanone [42] 256 256 5,7-dihydroxyflavanone [41] 242 dihydroxyflavan 272 7,3'-dihydroxy-4'-methoxyflavan [42] 256 256 7-hydroxy-3-(4- hydroxybenzyl)chroman 286 7-hydroxy-3-(4-hydroxybenzyl) - 8-methoxy-chroman 286 286 7-hydroxy-3-(3-hydroxy-4- methoxybenzyl)chroman 284 3-(4-hydroxybenzyl)-7,8- methylenedioxychroman 316? 316 [41], [42], [43] [41], [42] [41], [42] [41], [42], [43] 4/10

Table S3-b: Mass spectral data of compounds identified in dragon's blood of Dracaena cinnabari after methylation with TMSH. No. 10 most abundant MS peaks MW tentative assignment DrC-A 240 (100), 136 (35), 104 (32), 149 (29), 91 (27), 108 (25), 78 240 DrC1 + Me (24), 239 (22), 77 (19), 241 (18) DrC-B 151 (100), 254 (53), 272 (51), 121 (41), 77 (32), 165 (31), 223 272 DrC2 + Me (26), 140 (26), 152 (25), 105 (25) DrC-C 134 (100), 270 (64), 121 (41), 119 (16), 135 (15), 147 (12), 91 270 DrC7 + 2 Me (12), 271 (12), 239 (8), 146 (8) DrC-D 151 (100), 284 (28), 152 (11), 121 (9), 135 (8), 253 (8), 91 (6),? 285 (6), 316 (5), 165 (5) DrC-E 121 (100), 284 (79), 162 (44), 161 (29), 163 (26), 122 (20), 285 284 DrC10 + 2 Me (16), 176 (15), 175 (14), 147 (12) DrC-F 164 (100), 300 (79), 151 (47), 149 (28), 269 (18), 136 (17), 301 300 DrC8 + 2 Me (15), 121 (13), 165 (13), 177 (13) DrC-G 151 (100), 121 (21), 152 (13), 150 (12), 314 (11), 164 (10), 91 300 (9), 77 (5), 165 (5), 300 (4) DrC-H 151 (100), 300 (38), 135 (36), 138 (29), 121 (20), 165 (11), 77 300 (10), 152 (10), 301 (7), 91 (6) DrC-I 314 (100), 121 (62), 315 (21), 192 (19), 193 (17), 167 (16), 191 314 DrC11 + 2 Me (10), 147 (10), 122 (9), 161 (6) DrC-K 121 (100), 298 (45), 122 (25), 147 (19), 176 (17), 91 (12), 151 298 (12), 175 (10), 177 (10), 299 (9) DrC-L 152 (100), 314 (38), 151 (34), 121 (22), 137 (21), 161 (12), 153 314 DrC13 + 2 Me (10), 162 (9), 315 (8), 163 (6) DrC-M 181 (100), 151 (26), 330 (18), 135 (17), 121 (16), 168 (15), 165 330 (15), 182 (11), 77 (10), 136 (8) DrC-N 151 (100), 164 (56), 121 (23), 177 (16), 181 (15), 149 (15), 165 330 DrC16 + Me (13), 152 (13), 330 (13), 91 (10) DrC-O 121 (100), 298 (60), 165 (57), 283 (51), 135 (33), 122 (21), 161 298 DrC14 + Me (20), 270 (20), 297 (16), 133 (16) DrC-P 164 (100), 344 (27), 151 (25), 152 (24), 149 (17), 177 (14), 165 (11), 121 (10), 91 (9), 137 (7) 344 5/10

Table S4-a: Mass spectral data of compounds identified in dragon's blood of Dracaena draco. No. 10 most abundant MS peaks MW tentative assignment Ref. DrD1 120 (100), 242 (92), 123 (59), 107 (33), 91 (27), 136 (23), 65 (21), 119 (19), 133 (16), 121 (15) DrD2 120 (100), 137 (85), 256 (79), 107 (36), 121 (34), 91 (27), 150 (27), 77 (24), 133 (17), 119 (17) DrD3 150 (100), 272 (68), 135 (36), 137 (32), 77 (19), 107 (19), 123 (18), 136 (13), 65 (12), 273 (12) DrD4 150 (100), 286 (58), 137 (46), 107 (21), 131 (21), 135 (20), 163 (13), 149 (12), 255 (11), 151 (11) DrD5 107 (100), 256 (81), 148 (50), 147 (42), 77 (34), 149 (31), 108 (20), 123 (19), 133 (17), 257 (14) DrD6 150 (100), 272 (75), 135 (60), 107 (55), 137 (46), 77 (35), 147 (21), 162 (20), 123 (18), 65 (17) DrD7 107 (100), 286 (83), 77 (33), 178 (30), 147 (30), 153 (22), 133 (17), 179 (17), 177 (15), 287 (15) DrD8 150 (100), 286 (53), 137 (48), 135 (47), 77 (23), 107 (23), 162 (16), 151 (12), 163 (12), 79 (11) DrD9 284 (100), 137 (31), 285 (19), 115 (14), 131 (14), 128 (12), 138 (10), 147 (9), 69 (9), 77 (8) DrD10 107 (100), 284 (97), 150 (40), 176 (40), 151 (33), 77 (33), 147 (29), 133 (24), 91 (22), 175 (22) DrD11 121 (100), 284 (24), 138 (17), 137 (15), 77 (13), 122 (11), 91 (8), 153 (8), 78 (8), 147 (8) DrD12 153 (100), 272 (96), 107 (62), 120 (55), 300 (47), 133 (43), 121 (36), 69 (34), 192 (33), 138 (32) DrD13 166 (100), 286 (84), 167 (75), 120 (42), 136 (35), 107 (27), 133 (27), 137 (27), 148 (25), 91 (21) DrD14 286 (100), 107 (97), 178 (85), 177 (63), 179 (58), 77 (36), 153 (35), 69 (29), 287 (20), 108 (15) 242 dihydroxyflavan 256 4,7-dihydroxyflavanone or 4',7-dihydroxy-8-methylflavan [46] / [13], [46], [47] 272 4',7-dihydroxy-3'-methoxyflavan [13], [46], [47] 286 4',7-dihydroxy-3'-methoxy-8- methylflavan 256 7-hydroxy-3-(4- hydroxybenzyl)chroman 272 3',7-dihydroxy-4'-methoxyflavan 286 5,7-dihydroxy-3-(4- hydroxybenzyl)chroman-4-one 286 3',7-dihydroxy-4'-methoxy-8- methylflavan 284 10-hydroxy-11-methoxydracaenone 284 7,8-methylenedioxy-3-(4- hydroxybenzyl)chroman 284 300 5-7-dimethoxy-3-(4- hydroxybenzyl)chroman and other flavonoids 286 4',5-dihydroxy-7-methoxy-8- methylflavan 286 7-hydroxy-8-methoxy-3-(4- hydroxybenzyl)chroman [13], [46], [47] [13], [46] [13], [46] [46], [47] [46] [46] [46], [47] [13], [46] 6/10

Table S4-b: Mass spectral data of compounds identified in dragon's blood of Dracaena draco after methylation with TMSH. No. 10 most abundant MS peaks MW tentative assignment DrD-A 134 (100), 270 (84), 121 (36), 271 (15), 135 (15), 119 (14), 270 DrD1 + 2 Me 147 (11), 239 (11), 91 (10), 146 (8) DrD-B 134 (100), 284 (69), 121 (37), 119 (14), 135 (14), 91 (13), 284 DrD2 + 2 Me 285 (13), 147 (11), 253 (9), 120 (8) DrD-C 284 (100), 121 (53), 162 (22), 253 (20), 285 (19), 161 (17), 284 DrD5 + 2 Me 163 (15), 145 (15), 269 (10), 122 (10) DrD-D 164 (100), 300 (89), 151 (37), 149 (26), 269 (18), 301 (17), 300 DrD3 / DrD6 + 2 Me 136 (17), 177 (14), 165 (13), 121 (12) DrD-E 164 (100), 314 (98), 151 (35), 283 (23), 149 (22), 315 (21), 314 DrD4 / DrD8 + 2 Me 165 (15), 299 (14), 177 (14), 143 (12) DrD-F 314 (100), 121 (69), 180 (50), 315 (22), 147 (20), 192 (17), 314 193 (16), 162 (16), 167 (13), 134 (11) DrD-G 180 (100), 314 (88), 121 (29), 162 (25), 147 (23), 315 (19), 314 DrD13 + 2 Me 150 (18), 134 (17), 181 (13), 151 (10) DrD-H 121 (100), 314 (75), 192 (65), 328 (48), 193 (43), 191 (41), 314 DrD12 + Me 297 (33), 145 (20), 206 (18), 315 (17) DrD-I 121 (100), 298 (83), 283 (62), 165 (50), 135 (33), 270 (24), 122 (21), 161 (19), 297 (18), 299 (17) 298 DrD10 + Me 7/10

Table S5-a: Mass spectral data of compounds identified in dragon's blood of Daemonorops draco. No. 10 most abundant MS peaks MW tentative assignment Ref. DaeD1 256 (100), 152 (63), 117 (25), 257 (18), 124 (16), 165 (14), 153 (13), 91 (11), 239 (11), 225 (10) DaeD2 270 (100), 166 (51), 239 (19), 271 (19), 138 (189), 117 (14), 179 (13), 168 (13), 91 (12), 165 (10) DaeD3 167 (100), 272 (30), 137 (20), 105 (19), 168 (13), 77 (12), 152 (9), 256 (5), 273 (5), 69 (5) DaeD4 256 (100), 179 (80), 153 (63), 255 (62), 124 (31), 153 (18), 257 (15), 104 (15), 103 (14), 69 (12) DaeD5 256 (100), 284 (90), 165 (45), 152 (35), 213 (22), 91 (21), 137 (20), 257 (18), 90 (17), 285 (17) DaeD6 253 (100), 254 (91), 167 (39), 177 (26), 238 (25), 117 (20), 255 (15), 239 (11), 286 (11), 152 (10) DaeD7 268 (100), 267 (91), 252 (33), 191 (27), 253 (18), 269 (17), 237 (14), 251 (13), 118 (9), 77 (9) DaeD8 154 (100), 167 (68), 105 (32), 286 (29), 77 (23), 137 (20), 168 (14), 166 (10), 155 (10), 123 (8) DaeD9 134 (100), 286 (69), 147 (24), 121 (24), 119 (13), 287 (13), 135 (11), 91 (10), 152 (9), 146 (7) DaeD10 251 (100), 266 (68), 265 (30), 267 (29), 252 (23), 268 (18), 223 (14), 237 (13), 165 (12), 77 (11) 256 7-hydroxy-5-methoxyflavan [38], [50] 270 7-hydroxy-5-methoxy-6- methylflavan 272 256 284 286 268 286 4,6-dihydroxy-2-methoxy-3- methyldihydrochalcone 286 7-hydroxy-4',5-dimethoxyflavan [38], [50] [38] 266 dracorhodin [38], [50] Table S5-b: Mass spectral data of compounds identified in dragon's blood of Daemonorops draco after methylation with TMSH. No. 10 most abundant MS peaks MW tentative assignment DaeD-A 270 (100), 166 (62), 138 (25), 117 (22), 271 (19), 179 (15), 167 270 DaeD1 + Me (14), 239 (13), 154 (12), 253 (11) DaeD-B 284 (100), 180 (71), 152 (33), 285 (20), 253 (17), 117 (17), 193 284 DaeD2 + Me (16), 137 (13), 91 (13), 109 (11) DaeD-C 270 (100), 166 (81), 138 (28), 117 (27), 91 (27), 139 (24), 179 270 (20), 271 (19), 165 (17), 123 (17) DaeD-D 210 (100), 164 (94), 181 (88), 151 (42), 195 (30), 149 (21), 121 (19), 75 (17), 311 (16), 105 (15) DaeD-E 211 (100), 181 (40), 296 (25), 195 (14), 212 (13), 210 (12), 105 (11), 77 (11), 297 (10), 151 (9) DaeD-F 134 (100), 300 (91), 147 (37), 166 (36), 121 (31), 301 (17), 138 (15), 135 (13), 119 (13), 146 (12) 300 DaeD9 + Me 8/10

Table S6-a: Mass spectral data of compounds identified in dragon's blood of Daemonorops micracantha. No. 10 most abundant MS peaks MW DaeM1 268 (100), 267 (96), 252 (31), 191 (25), 253 (19), 269 (17), 237 (15), 251 (13), 118 (9), 223 (6) DaeM2 134 (100), 286 (66), 147 (25), 121 (23), 119 (12), 287 (12), 135 (11), 91 (9), 152 (8), 146 (7) DaeM3 272 (100), 153 (97), 120 (35), 133 (33), 273 (17), 152 (15), 107 (15), 69 (12), 165 (12), 132 (12) DaeM4 150 (100), 316 (94), 302 (66), 167 (64), 153 (44), 137 (37), 135 (29), 163 (25), 151 (20), 317 (19) DaeM5 316 (100), 164 (66), 150 (29), 167 (27), 317 (19), 151 (18), 177 (18), 149 (17), 165 (15), 285 (13) DaeM6 283 (100), 284 (93), 268 (31), 269 (15), 253 (11), 267 (11), 285 (10), 126 (9), 191 (8), 142 (7) DaeM7 316 (100), 150 (84), 167 (72), 137 (49), 131 (22), 317 (21), 163 (20), 135 (20), 165 (19), 151 (16) 268 286 272 316 316 284 316 Table S6-b: Mass spectral data of compounds identified in dragon's blood of Daemonorops micracantha after methylation with TMSH. No. 10 most abundant MS peaks MW tentative assignment DaeM-A 300 (100), 134 (82), 166 (34), 147 (33), 121 (25), 301 300 DaeM2 + Me (20), 138 (13), 269 (10), 135 (10), 146 (10) DaeM-B 164 (100), 330 (83), 151 (32), 177 (27), 149 (22), 331 330 DaeM4 + Me (19), 176 (16), 165 (16), 299 (15), 315 (14) DaeM-C 164 (100), 344 (87), 151 (25), 345 (20), 177 (18), 149 344 DaeM5 + 2 Me (17), 165 (15), 313 (12), 195 (12), 152 (11) DaeM-D 327 (100), 175 (73), 176 (55), 151 (36), 343 (35), 145 (33), 328 (24), 179 (21), 177 (21), 161 (20) 328 (?) 9/10

Table S7: Triterpenoids identified in some Daemonorops draco resins of the historic Martius collection. In the derivatised extract, additional triterpenoid acids can be identified that were also found before [37]. The compounds marked in grey are typical for dammar resin. No. 10 most abundant MS peaks MW tentative assignment Ref. T1 109 (100), 205 (66), 95 (61), 69 (51), 204 (48), 189 (48), 93 (47), 108 (45), 107 (38), 81 (34) T2 218 (100), 203 (61), 189 (26), 205 (22), 109 (21), 95 (20), 219 (18), 69 (17), 204 (15), 107 (14) T3 109 (100), 189 (89), 207 (76), 204 (65), 95 (65), 69 (56), 93 (55), 108 (54), 135 (52), 107 (45) T4 204 (100), 134 (33), 175 (27), 191 (26), 189 (25), 205 (25), 95 (25), 122 (24), 109 (22), 135 (22) T5 218 (100), 204 (39), 203 (31), 189 (26), 95 (25), 219 (23), 109 (22), 205 (21), 122 (21), 107 (19) T6 205 (100), 189 (74), 121 (61), 109 (56), 95 (53), 123 (47), 204 (45), 122 (43), 107 (43), 425 (41) T7 109 (100), 205 (47), 95 (46), 69 (42), 356 (35), 425 (31), 93 (29), 135 (24), 107 (24), 123 (23) T8 203 (100), 204 (33), 232 (26), 189 (19), 205 (17), 109 (13), 119 (13), 95 (12), 175 (12), 81 (12) T9 203 (100), 133 (37), 204 (22), 205 (17), 232 (15), 119 (10), 105 (8), 189 (8), 95 (8), 134 (7) T10 203 (100), 133 (33), 204 (21), 232 (15), 189 (14), 207 (12), 119 (12), 190 (11), 95 (10), 105 (10) 424 Dammaradienone [55]?? 426 Dammaradienol [55] 410 Nor-α-Amyrone [55] 426 α-amyrin 442? 442 Hydroxydammarenone [55], [37] 438 Oleanonic aldehyde [55], [37] 438 Ursonic aldehyde [55], [37]? 10/10