Identification and Quantification of Feathers, Down, and Hair of Avian and Mammalian Origin Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Abstract

We describe a fast method for the identification of the origin of native and chemically processed feathers, down, and hair and for distinguishing closely related species using enzyme digestion and MALDI-TOF mass spectrometry. Additionally we present two methods for the quantification of different identified bird and mammalian samples, respectively, in binary species mixtures. Without any prior cleaning or isolation of single proteins, enzymatical digests of feathers, down, and hair are performed. Fragments generated are analyzed by MALDI-TOF mass spectrometry, and peak groups of different selectivity are established for every animal species. For the identification of individual animal species, only unique species-specific peaks are accepted while for species classification of tinted or bleached hair, the group of semispecific peaks (SEMPs) is used. Samples from native animal species show a higher coincidence of digest peak masses of SEMPs than far-related species, indicating a phylogenetical relationship of the investigated structure proteins. Quantification of mixed binary avian samples is performed by identification of preweighed single feathers and down, followed by calculation of the gravimetric mass fractions. The composition of mixed binary mammalian samples is calculated from correlation of the quotient of the relative peak intensities or areas of these species and the quotient of corresponding gravimetric mass fractions. High accuracy is achieved by both quantification methods. The fast methods are well suited for industrial quality control for example.

Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry combined with multidimensional scaling, binary hierarchical cluster tree and selected diagnostic masses improves species identification of Neolithic keratin sequences from furs of the Tyrolean Iceman Oetzi

“…[34,45] Species-specific amino acid sequences in combination with extraordinary robustness explicated above [35,36] make hair keratins ideal for species identification. [46][47][48] For trypsin collagen digests a very similar species identification method is described based on species-specific amino acid sequences and the extraordinary robustness of collagen in bones. [49] Keratin-based species identification method Ancient proteins extracted and sequenced from fossil bones offer an extraordinary reservoir of genetic information that has recently become tractable through the application of mass spectrometric (MS) techniques.…”

“…The keratin-based method used in this study (SIAM = Species Identification of Animals using MALDI-TOF mass spectrometry) was developed for the identification of native feathers, down and hair. [14,46,47] Pmfs from digests of intact hair show species-specific differences in their peptide patterns. Specific calculation of their spectral similarities is a measure for their zoological relationship shown by subsequent graphic visualization routines.…”

Differential Identification and Quantification of Cashmere, Sheep and Yak Fibers in Textiles Using Liquid Chromatography/Mass Spectrometry

“…Matrix-assisted laser desorption/ionization-time-offlight mass spectrometer (MALDI-TOFMS) was first studied as an instrument to identify animal fibers [6][7][8][9][10]. These methods rely either on the comparison of patterns of peptides [6][7][8] or on the identification of speciesspecific peptides [9,10].…”

“…These methods rely either on the comparison of patterns of peptides [6][7][8] or on the identification of speciesspecific peptides [9,10]. The methods using MALDI-TOFMS are generally rapid and offer high operational throughput.…”

Peptide Profiling Using Matrix-Assisted Laser Desorption/Ionization-Time-of-Flight Mass Spectrometry for Identification of Animal Fibers

“…We have previously studied the identification and qualification of cashmere and wool (untreated and dyed) fibers using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOFMS), 3) with a similar study focusing on feathers and down. 4) Proteome analysis of cashmere, 5) and identification of cashmere, wool, and yak fibers by peptide mass fingerprint analysis have also been conducted. 6,7) To date, identification of animal fibers has been mostly applied to untreated cashmere, wool, and yak; however, in order to accurately differentiate cashmere from substituted fibers of lower value, it is necessary to widen the scope of materials included in the investigations.…”