# Organic Materials Used for Giant Buddhas and Wall Paintings in Bamiyan, Afghanistan
## 3 Results
### Identification of Protein and Animal Sources of Glue by LC-MS
#### 3.5.2 Species Identification for Glue
\begin{table}
\begin{tabular}{l c c c c} \hline \hline _Peptide (COLIA1)_ & _m/z_ & _Mass (Da)_ & _Sample (BMM)_ & _Species_ \\ \hline GEAGPSGPAGPTGAR & 641.6 & 1281.2 & 191, 201 & _Bos_ \\ \hline GPSGPQGPSGPOGPK & 666.7 & 1331.4 & 199, 201 & _Bos_ \\ \hline GPSGPqGPSGPOGPK & 667.2 & 1332.4 & 191, 201 & _Bos_ \\ \hline GEOGPAGLPGOOGER & 718.4 & 1434.8 & 191, 201 & _Bos_ \\ \hline GAAGPOGATGFOGAAGR & 730.6 & 1546.2 & 191, 201 & _Bos_ \\ \hline GETGPGPAGPGVGPVAR *1 & 774.1 & 1545.8 & 191, 201 & _Sus_ \\ \hline GETGPAGPAGPIGPVGAR & 781.3 & 1560.5 & 191, 199, 201 & _Bos_ \\ \hline DEAGAAGGPPOAGPAGER & 854.5 & 1706.9 & 199, 201 & _Bos_ \\ \hline GEPGSGGenGAPGQmGOR & 880.8 & 1759.6 & 199, 201 & _Bos_ \\ \hline SORDGETGPAGPIGPVGAR & 659.7 & 1975.0 & 199, 201 & _Bos_ \\ \hline GAPGADGPGAOGTPGPQIGAGR & 1037.9 & 2073.7 & 201 & _Bos_ \\ \hline GnDGATGAGOPGTGGPGPGPGAGVGAK & 850.8 & 2549.3 & 199, 201 & _Bos_ \\ \hline GFSCLQGPPGPOGSOGEqGPSGAGPAGPR & 897.6 & 2689.8 & 201 & _Bos_ \\ \hline GFSCLqGPGOOOGPSPEqGPSGASGPAGPR & 903.5 & 2707.7 & 191 & _Bos_ \\ \hline
**GLTGPIGPPGAGAPGDKGEAGPSGPAGPTGAR** & 952.0 & 2852.9 & 191, 201 & _Bos_ \\ \hline _Peptide (COLIA2)_ & _m/z_ & _Mass (Da)_ & _Sample (BMM)_ & _Species_ \\ \hline
**IGQPGAVGPAGIR** & 597.1 & 1192.1 & 199 & _Bos_ \\ \hline
**IGQOGAVGPAGIR** & 605.2 & 1208.3 & 191 & _Bos_ \\ \hline GEAGPAGPAGPAGPAGPR & 631.6 & 1261.2 & 191, 201 & _Bos_ \\ \hline GIOKGPAGAAGATG *1 & 620.6 & 1239.2 & 201 & _Sus, Equus_ \\ \hline GIGCPVGAAAGATG & 634.5 & 1267.1 & 191 & _Bos_ \\ \hline GPOGESGAAGPTGPIGSR & 791.2 & 1580.5 & 191, 199, 201 & _Bos_ \\ \hline GELGPVGnOGPAGPAGPR & 809.1 & 1616.2 & 191, 201 & _Bos_ \\ \hline GSTGEGPAGPOGPGLR & 825.3 & 1648.6 & 199, 201 & _Bos_ \\ \hline GLOGVAGSVGPGOLGIAGPOGAR & 1066.3 & 2130.5 & 191, 199, 201 & _Bos_ \\ \hline
**GYOGNAGPVGAAGAGPGOQGPVGPVGK** & 1131.4 & 2260.7 & 191, 199 & _Bos_ \\ \hline
**GYOGAGPVGAAGAPGOQGPVGPVGK** & 1132.9 & 2263.8 & 201 & _Bos_ \\ \hline GEqGOAGPPGFQGLPGOAGTAGEAGKOGER & 932.0 & 2792.3 & 199, 201 & _Bos_ \\ \hline _Peptide (COLIA1)_**22** & _m/z_ & _Mass (Da)_ & _Sample (BMM)_ & _Species_ \\ \hline
**GPOPOGGTnGVOGqR** & 719.7 & 1437.3 & 201 & _Bos_ \\ \hline GETGPAGPSGAOGPAGSR & 770.6 & 1539.3 & 201 & _Bos_ \\ \hline GEVPGAGSOGSSAOGqR & 800.3 & 1598.5 & 201 & _Bos_ \\ \hline GPOGPQLOGLAGTAGEOCR & 917.7 & 1833.4 & 201 & _Bos_ \\ \hline \hline \end{tabular} Abbreviations: We used a standard one—letter code for proteinaceous amino acids. The other codes for a few modification products are as follows: O, hydroxy—Pro; m, Met (M)—sulfoxide; q, deamidated Gln (Q); n, deamidated Asn (N). Amino acid residues distinguishing between _Bos (Bos taurus)_ and other animal species (_Sus: Sus scrafi; Equus: Equus caballus_) are printed in red. Sequences unique to deer, goats, sheep, pigs, and horses are printed in bold face. *1 Further analysis of these peptides is needed to confirm the identity of the species. Residues distinguishing between cattle and other species are printed in red. *2 See also Figure 6 for a comparison of sequences with respect to cattle, deer, goat, sheep, and horse.  
\end{table} 
Table 9: Trypic peptides identified originating from the type I collagen \(\alpha\)1-chain (COL1A1), \(\alpha\)2-chain (COL1A2), and type III collagen \(\alpha\)1-chain (COL3A1) in BMM191, BMM199, and BMM201. Although peptides identified from type III collagen were detected only in the sample BMM201, these peptides showed more clearly that the source of animal glue used as a binder was cattle hide. As shown in Figure 8, the amino acid sequences of peptides are not consistent with those of animals, including horses, pigs, sheep, goats, and deer, in line with the results obtained from type I collagen (Table 9). Moreover, the possibility of animal bone as a source of animal glue is unlikely because type III collagen is much less abundant in bones and tendons than type I collagen [46]. The full lists of MASCOT search results of peptides are summarized in Figures S2-S4 in Supporting Information.