Supporting Information

Identification of cross-species marker peptides for the detection of

mammalian and poultry meat in vegan and vegetarian foods

Lukas Häfner,1 Jens Brockmeyer,2 Ilka Haase,1 Bertolt Kranz,1 and Wolfgang Jira3,*

1
National Reference Centre for Authentic Food, Max Rubner-Institut (MRI), E.-C.-Baumann-Straße 20,

95326 Kulmbach, Germany

2
Department of Food Chemistry, Institute for Biochemistry and Technical Biochemistry, University of

Stuttgart, Allmandring 5B, 70569 Stuttgart, Germany

3
Department of Safety and Quality of Meat, Max Rubner-Institut (MRI), E.-C.-Baumann-Straße 20,

95326 Kulmbach, Germany

* Corresponding author. E-mail address: wolfgang.jira@mri.bund.de

1
Table S1: Sample materials (n = 51) of mammalian (n = 24) and poultry (n = 27) meat. The entries

marked gray (n = 15) were used for peptide identification (Section 3.2.), those marked dark gray (n = 7)

also for peptide selection (Section 3.4.). For homology testing, all of the sample materials were used

(Section 3.7.). a = for EPI scan (Section 3.4.); b = for spiking experiments (Section 3.5.).

Species Sample material Acquisition date

Pork steak (back) b 03/16/2017
Pig
Pork schnitzel (thigh) a,b 04/15/2020
(Sus scrofa)
Fat-reduced minced pork 01/11/2021
Beef goulash 04/15/2020
Cattle
Beef steak from young bull 03/16/2017
(Bos taurus)
Organic minced beef 01/11/2021
Deboned lamb back 01/11/2021
Sheep
New Zealand Lamb fillet 02/28/2022
(Ovis aries)
Lamb fillet 02/28/2022
Goat meat 03/25/2021
Goat
Minced goat meat 07/29/2022
(Capra hircus)
Flank steak from young kid 07/29/2022
Rabbit fillet (back) 01/11/2021
Rabbit
Rabbit leg 02/08/2022
(Oryctolagus cuniculus)
Rabbit parts (back, leg) 02/08/2022
Horse roulade (leg) 11/29/2021
Horse
Horse sauerbraten (shoulder) 02/12/2022
(Equus caballus)
Horse sauerbraten (eye of round steak) 02/12/2022
Red deer goulash 01/11/2021
Red deer
Red deer goulash 02/28/2022
(Cervus elaphus)
Red deer medallions (leg) 02/28/2022
Roe deer medallions 01/11/2021
Roe deer
Roe deer goulash 02/28/2022
(Capreolus capreolus)
Roe deer medallions (leg) 02/28/2022
Chicken schnitzel b 01/16/2020
Chicken
Chicken breast fillet a,b 01/16/2020
(Gallus gallus)
Chicken breast fillet 05/09/2019
Turkey schnitzel 05/09/2019
Turkey
Turkey schnitzel 01/16/2020
(Meleagris gallopavo)
Turkey steak (breast) 01/16/2020
Duck Barbarie duck breast fillet 05/09/2019
(Anas platyrhynchos, Barbarie duck breast 09/11/2019
Cairina moschata) Duck breast fillet 01/16/2020
Goose Goose breast fillet 05/09/2019
(Anser anser, Goose breast 09/11/2019
Anser cygnoides) Polish goose breast 09/11/2019

2
Table S1: Continued.

Homogenized pheasant meat 06/04/2019

Pheasant
Wild pheasant (whole) 09/20/2019
(Phasianus colchicus)
Pheasant breast fillet 02/11/2020
Homogenized quail meat 06/04/2019
Quail
Quail (whole) 09/20/2019
(Coturnix japonica)
Quail (whole) 02/14/2020
Homogenized Guinea fowl meat 06/04/2019
Guinea fowl
Guinea fowl (whole) 09/20/2019
(Numida meleagris)
Guinea fowl breast 02/10/2020
Ostrich steak 01/23/2020
Ostrich
Ostrich meat strips [note: pet food] 02/07/2020
(Struthio camelus)
Ostrich steak 04/06/2020
Wild pigeon breast 09/20/2019
Pigeon
Pigeon (whole) 02/14/2020
(Columba livia)
Domestic pigeon 04/06/2020

3
Table S2: Base materials and ingredients of the three main components (matrix, textured pea protein,

emulsifier) and the seasoning for the commercial vegan bratwurst-style sausages. The complete recipe

is a property of Van Hees (Walluf, Germany). The pork and chicken meat (n = 1, each) for the spiking

of the vegan sausages was acquired from local supermarkets in the form of whole muscle pieces/cuts

and minced in-house prior to use.

Matrix
Methylcellulose (E461)
Base material PRALLO® Matrix V
Potato starch
Rapeseed oil
Ingredients
Ice
Texturized pea protein
Pea protein
Base material PRALLO® Struktur veggian AF
Pea flour
Ingredients Water
Emulsifier
Processed Eucheuma algae (E407a)
Potato starch
Pea protein
Base material PRALLO® Funktion veg. Bratwurst
Table salt
Citrus fiber
Rapeseed oil
Rapeseed oil
Ingredients
Ice
Seasoning
Base material VAN HEES® Würzung veg. Bratwurst mA
Potato starch
Table salt
Ingredients
Marjoram
Citric acid

Species Sample material Acquisition date

Pig (Sus scrofa) Pork shoulder 06/14/2021
Chicken (Gallus gallus) Chicken breast fillet 07/19/2021

4
Table S3: Sixteen different groups of 136 food-relevant, plant-based (or plant-like) sample materials.

The groups represent categories of related foods, selected based on the European vegetable production

in 2019 (≥ 100,000 metric tonnes after grouping) provided by the Food and Agriculture Organisation of

the United Nations;a data obtained from the German Federal Ministry of Food and Agriculture about the

vegetable consumption in Germany between 2018 and 2019;b a review of the scientific literature

regarding meat substitutes; and a survey of the ingredients of commercial vegan and vegetarian foods.

Another five sample materials were chosen either based on suspected homologies of the marker peptide

IDFDEFLK (Section 3.3.) or to exclude homologies in the sample preparation control (alfalfa).

Group 1: Common Cereals Group 2: Millets

Wheat (Triticum aestivum) Proso millet (Panicum miliaceum)
Spelt (Triticum spelta) Golden millet (Panicum miliaceum)
Rye (Secale cereale) Silver millet (Panicum miliaceum)
Triticale (× Triticosecale Wittmack) Pearl millet (Pennisetum glaucum)
Barley (Hordeum vulgare) Red Foxtail millet (Setaria italica)
Oat (Avena sativa) Yellow Foxtail millet (Setaria italica)
Rice (Oryza sativa) Sorghum (Sorghum bicolor)
Maize (Zea mays) Teff (Eragrostis tef)
Group 3: Pseudo Cereals and Substitutes Group 4: Tubers and Bulb Vegetables
Buckwheat (Fagopyrum esculentum) Potato (Solanum tuberosum)
Quinoa (Chenopodium quinoa) Sweet potato (Ipomoea batatas)
Amaranth (Amaranthus spp.) Cassava (Manihot esculenta)
Chia (Salvia hispanica) Jerusalem artichoke (Helianthus tuberosus)
Flax (Linum usitatissimum) Ginger (Zingiber officinale)
Sweet chestnut (Castanea sativa) Celeriac [root]
(Apium graveolens var. rapaceum)
Fennel [bulb] (Foeniculum vulgare)

5
Table S3: Continued.

Group 5: Root Vegetables Group 6: Onions and Leaks

Turnip (Brassica rapa ssp. rapa var. majalis) Onion (Allium cepa var. cepa)
Rutabaga (Brassica napus ssp. rapifera) Shallot (Allium cepa var. aggregatum)
Beetroot Spring onion (Allium fistulosum)
(Beta vulgaris ssp. vulgaris Conditiva Group) Leek (Allium ampeloprasum Leek Group)
Mangelwurzel Garlic (Allium sativum)
(Beta vulgaris ssp. vulgaris Crassa Group) Wild garlic (Allium ursinum)
White radish
(Raphanus sativus convar. sativus
Radish Group)
Red radish
(Raphanus sativus convar. sativus
Small Radish Group)
Carrot (Daucus carota)
Parsnip (Pastinaca sativa)
Parsley [root] (Petroselinum crispum)
Horseradish (Armoracia rusticana)
Group 7: Salads Group 8: Leaf Vegetables
Butterhead lettuce Spinach (Spinacia oleracea)
(Latuca sativa Butterhead Group) Pak choi (Brassica rapa ssp. chinensis)
Iceberg lettuce Napa cabbage
(Latuca sativa Crisphead Group) (Brassica rapa ssp. pekinensis)
Chicory Celery [leaf]
(Cichorium intybus var. foliosum (Apium graveolens var. secalinum)
Foliosum Group) Artichoke
Radicchio (Cynara cardunculus Scolymus Group)
(Cichorium intybus var. foliosum
Radicchio Group)
Rucola (Eruca vesicaria)
Common cornsalad (Valerianella locusta)
Chard
(Beta vulgaris ssp. vulgaris Cicla Group)

6
Table S3: Continued.
Group 9: Cabbages
Kale (Brassica oleracea var. sabellica)
White cabbage
(Brassica oleracea convar. capitata var. alba)
Red cabbage
(Brassica oleracea convar. capitata var. rubra)
Savoy cabbage
(Brassica oleracea convar. capitata
var. sabauda)
Broccoli (Brassica oleracea var. italica)
Cauliflower (Brassica oleracea var. botrytis)
Brussel sprouts
(Brassica oleracea var. gemmifera)
Kohlrabi
(Brassica oleracea var. gongylodes)
Group 11: Beans
Garden bean (Phaseolus vulgaris ssp.)
Kidney bean (Phaseolus vulgaris ssp.)
Pinto bean (Phaseolus vulgaris ssp.)
Black bean (Phaseolus vulgaris ssp.)
White bean (Phaseolus vulgaris ssp.)
Runner bean (Phaseolus coccineus ssp.)
Giant bean (Phaseolus coccineus ssp.)
Lima bean (Phaseolus lunatus)
Broad bean (Vicia faba ssp. faba var. minuta)
Mung bean (Vigna radiata)
Group 13: Nuts
Walnut (Juglans regia)
Hazelnut (Corylus avellana)
Almond (Prunus dulcis)
Cashew (Anacardium occidentale)
Macadamia (Macadamia spp.)
Coconut (Cocos nucifera)
Brazil nut (Bertholletia excelsa)
Pecan nut (Carya illinoinensis)
Pistachio (Pistacia vera)
Pine seed (Pinus pinea)
7
Group 10: Fruit Vegetables
Tomato (Solanum lycopersicum)
Eggplant (Solanum melongena)
Bell pepper (Capsicum annuum)
Chili pepper (Capsicum frutescens)
Habanero (Capsicum chinense ‘Habanero’)
Cucumber (Cucumis sativus)
Pumpkin [seed protein] (Cucurbita spp.)
Zucchini
(Cucurbita pepo var. cylindrica
Zucchini Group)
Avocado (Persea americana)
Olive (Olea europaea)
Group 12: Other Legumes
Soy bean (Glycine max)
Pea (Pisum sativum)
Chick pea (Cicer arietinum)
Brown lentil (Lens culinaris ssp.)
Red lentil (Lens culinaris ssp.)
White lupin (Lupinus albus)
Blue lupin (Lupinus angustifolius)
Yellow lupin (Lupinus luteus)
Peanut (Arachis hypogaea)
Group 14: Selected Fruits and Thickeners
Jackfruit (Artocarpus heterophyllus)
Blackcurrant (Ribes nigrum)
Red Kuri squash [fruit]
(Cucurbita maxima ‘Hokkaido’)
Red Kuri squash [seed]
(Cucurbita maxima ‘Hokkaido’)
Apple (Malus domestica)
Sweet orange [peel] (Citrus × sinensis)
Lemon [peel] (Citrus limon)
Banana (Musa × paradisiaca)
Locust bean gum (Ceratonia siliqua)
Psyllium husk (Plantago spp.)
Table S3: Continued.

Group 15: Algae and Mushrooms Group 16: Others

Nori (Porphyra spp.)
Sea spaghetti (Himanthalia elongata)
Champignon (Agaricus bisporus)
Porcino (Boletus edulis)
Chanterelle (Cantharellus cibarius)
King trumpet mushroom (Pleurotus eryngii)
Oyster mushroom (Pleurotus ostreatus)
Shiitake (Lentinula edodes)
Quorn® (Fusarium venenatum)
Baker’s yeast (Saccharomyces cerevisiae)
Sunflower [seed protein] (Helianthus annuus)
Pumpkin [seed] (Cucurbita spp.)
Asparagus (Asparagus officinalis)
Bamboo [shoot] (Bambusoideae)
Breadseed poppy (Papaver somniferum)
Sesame (Sesamum indicum)
White mustard (Sinapis alba)
Black mustard (Brassica nigra)
Brown mustard (Brassica juncea)
Hemp [protein] (Cannabis sativa)

Additional Sample Materials

Pigeon pea (Cajanus cajan)
Taro (Colocasia esculenta)
Sunflower [seed] (Helianthus annuus)
Sweet orange [whole fruit] (Citrus × sinensis)
Alfalfa (Medicago sativa)

a Food and Agriculture Organisation of the United Nations (FAO). FAOSTAT. Crops and

livestock products https://www.fao.org/faostat/en/#data/QCL (accessed Jan 28, 2021).

b German Federal Ministry of Food and Agriculture (BMEL). Konsum von Gemüse in

Deutschland nach Art in den Jahren 2017/18 und 2018/19 (in 1.000 Tonnen)

https://de.statista.com/statistik/daten/studie/215858/umfrage/marktverbrauch-von-gemuese-

nach-arten-in-deutschland/ (accessed Jan 28, 2021).

8
Table S4: Sample materials (n = 18) of commercial spray-dried milk (n = 3), egg (n = 3), casein (n = 3),

whey (n = 3), albumen (n = 3), and yolk (n = 3), purchased from seven different producers.

Designation Product description Producer Acquisition date

Milk 1 Organic whole milk powder A 10/05/2021
Milk 2 Whole milk powder B 10/06/2021
Milk 3 Skim milk powder B 10/06/2021
Egg 1 Organic whole egg powder A 10/05/2021
Egg 2 Whole egg powder B 10/06/2021
Egg 3 Whole egg powder C 02/03/2021
Casein 1 Organic micellar casein D 10/05/2021
Casein 2 Micellar casein E 03/30/2020
Casein 3 Casein from bovine milk [for laboratory use] F 09/12/2022
Whey 1 Organic whey protein D 10/05/2021
Whey 2 Whey protein E 03/30/2020
Whey 3 Whey from bovine milk [for laboratory use] F 08/31/2022
Albumen 1 Albumen powder G 10/05/2021
Albumen 2 Albumen powder B 10/06/2021
Albumen 3 Egg whites from chicken [for laboratory use] F 08/15/2021
Yolk 1 Yolk powder G 10/05/2021
Yolk 2 Yolk powder C 02/24/2020
Yolk 3 Egg yolks from chicken [for laboratory use] F 08/15/2022

9
Table S5: Sample materials (n = 19) of commercial vegetarian meat substitute products based on
milk (n = 6; marked gray) and egg (n = 13; marked dark gray), purchased from eight different producers
(acquisition date: 11/09/2022).
Product description
Vegetarian schnitzel
with gouda cheese
Vegetarian schnitzel
Vegetarian geschnetzeltes
Vegetarian steak
Vegetarian nuggets
Vegetarian crispy sticks
Vegetarian mini-bratwurst
Vegetarian hot dogs
Vegetarian (vienna) sausages
Vegetarian (vienna) sausages
Vegetarian (vienna) sausages
Vegetarian bologna sausage
Vegetarian cold cuts
(chicken breast)
Vegetarian snack-salami
Vegetarian mini-salami
Vegetarian mini vienna
sausages
Vegetarian vienna sausages
Vegetarian frikadeller
Vegetarian (meat) balls
Ingredients derived from milk and egg Producer
50% skim milk, 14% gouda cheese,
chicken egg protein [percentage undisclosed],
cheddar cheese powder [percentage undisclosed],
flavor (derived from milk) [percentage undisclosed]
64% skim milk,
chicken egg protein [percentage undisclosed],
flavor (derived from milk) [percentage undisclosed]
77% skim milk,
chicken egg protein [percentage undisclosed],
flavor (derived from milk) [percentage undisclosed] H
76% skim milk,
chicken egg protein [percentage undisclosed]
62% skim milk,
chicken egg protein [percentage undisclosed],
flavor (derived from milk) [percentage undisclosed]
56 % skim milk,
chicken egg protein [percentage undisclosed],
cheddar cheese powder [percentage undisclosed],
flavor (milk-derived) [percentage undisclosed]
9% chicken egg protein powder I
5.3% egg protein powder J
10% chicken egg albumen powder K
10% chicken egg protein L
10% chicken egg protein powder
9% chicken egg protein powder
M
5% chicken egg protein powder
Dried egg albumen [percentage undisclosed]
Dried egg albumen [percentage undisclosed]
8% dried egg albumen N
8% dried egg albumen
Dried egg albumen [percentage undisclosed]
Mayonnaise
(with chicken egg yolk [percentage undisclosed]),
O
chicken egg protein powder
[percentage undisclosed]
10
Table S6: Parameters of the LC run for HR-MS/MS as well as the non-optimized and optimized LC runs

for LR-MS/MS. The mobile phase consisted of solvent A (water with 3% ACN and 0.1% FA) and

solvent B (water with 90% ACN and 0.1% FA).

LC run for HR-MS/MS

Time [min] [% B] Flow rate [μL/min]
0 2 250
3 2 250
33 60 250
34 100 250
44 100 250
45 2 250
52 2 250
Total run time: 52 min
Non-optimized LC run for LR-MS/MS
Time [min] [% B] Flow rate [μL/min]
0 10 250
1 10 250
26 30 250
27 100 250
33 100 250
34 10 250
39 10 250
Total run time: 39 min
Optimized LC run for LR-MS/MS
Time [min] [% B] Flow rate [μL/min]
0 10 250
1 10 250
16 40 250
16 40 500
17 100 500
22 100 500
22 10 500
27 10 500
27 10 250
28 10 250
Total run time: 28 min

11
Table S7: Amino acid sequences, retention times (tR), precursor ions, fragment ions, and MRM parameters of the 43 peptides preselected as meat markers

under non-optimized (single or upper row) and optimized (lower row) conditions. The five fragment ions of each peptide are listed in order of decreasing intensity

as observed by MRM. a = No fragment ions detectable by EPI scan; b = Fragment ion not detectable by non-optimized MRM; marked bold: first = quantifier,

second = qualifier 1, third = qualifier 2. The entries marked gray showed high signal intensities (least intense fragment ion) and good analytical performances in

seven food-important types of meat applying non-optimized MRM. DP = Declustering potential; CE = Collision energy; CXP = Collision cell exit potential. The tR

were derived from the seven meat matrices (non-optimized LC conditions) and from the vegan sausages spiked with 5%, w/w pork or 5%, w/w chicken meat

(optimized LC conditions) using the (preliminary) optimized MRM, and were confirmed with synthetic peptides. The entries 17 and 19 occurred only in mammals,

the entries 15 and 36 were not detectable (only their near-identical L/I-isomers).

m/z Precursor
No. Peptide sequence tR [min] DP [V] m/z Fragments (identity) CE [V] CXP [V]
(charge state)

1 YGYTHLSTGDLLR 14.1 499.3 (+3) 30 761.4 (y7)/401.3 (y3)/573.3 (y5)/666.8 (y12 2+)/638.3 (y112+) 30/30/30/30/30 36/36/36/36/36

2 GNPTVEVDLHTAK 6.3 690.9 (+2) 30 605.3 (y112+)/912.5 (y8)/783.4 (y7)/319.2 (y3)/570.3 (a6) 30/30/30/30/30 36/36/36/36/36

3 IEEALGDK 42 2.2 437.7 (+2) 30 319.2 (y3)/432.2 (y4)/632.3 (y6)/761.4 (y7)/216.6 (y4 2+) b 30/30/30/30/30 36/36/36/36/36

4 TDLNHENLK 1.6 542.3 (+2) 30 640.3 (y5)/434.2 (y72+)/503.3 (y4)/374.2 (y3)/260.2 (y2) 30/30/30/30/30 36/36/36/36/36

5 GILAADESTGSIAK 8.7 666.9 (+2) 30 284.2 (b3)/355.2 (b4)/581.8 (y12 2+)/978.5 (y10)/907.4 (y9) 30/30/30/30/30 36/36/36/36/36

6.7 685.4 (+2) 30 702.4 (y8)/815.5 (y9)/555.3 (b6)/668.4 (b7)/928.5 (y10) 30/30/30/30/30 36/36/36/36/36
6 GAAQNIIPASTGAAK 42
- 685.4 (+2) 111 702.4 (y8)/815.5 (y9)/555.3 (b6)/328.2 (b4)/289.2 (b3) 29/25/27/31/41 42/48/30/20/10
18.6 597.3 (+2) 30 686.4 (y6)/235.1 (a2)/377.1 (b3)/589.4 (y5)/931.5 (y8) 30/30/30/30/30 36/36/36/36/36
7 DFNMPLTISR
11.40 597.3 (+2) 81 686.4 (y6)/235.1 (a2)/377.1 (b3)/508.2 (b4)/262.2 (y2) 25/43/21/19/21 32/20/20/26/24

12
Table S7: Continued.

8 DSALSTHKPIEVK 2.7 712.9 (+2) 30 246.2 (y2)/585.4 (y5)/655.4 (y12 2+)/840.4 (b8)/387.2 (b4) 30/30/30/30/30 36/36/36/36/36

9 LQDAEEHVEAVNAK 2.1 776.9 (+2) 30 332.2 (y3)/631.3 (y6)/502.3 (y5)/357.2 (b3)/922.4 (b8) 30/30/30/30/30 36/36/36/36/36

10 LQNEVEDLMIDVER 42 24.1 851.9 (+2) 30 518.3 (y4)/762.4 (y6)/631.3 (y5)/403.2 (y3)/875,5 (y7) 30/30/30/30/30 36/36/36/36/36

11 ADIAESQVNK 1.5 537.8 (+2) 30 775.4 (y7)/704.4 (y6)/575.3 (y5)/261.2 (y2)/371.2 (b4) 30/30/30/30/30 36/36/36/36/36

12 AEDEEEINAELTAK 8.6 781.4 (+2) 30 746.4 (y7)/319.2 (y3)/859.5 (y8)/316.1 (b3)/988.5 (y9) 30/30/30/30/30 36/36/36/36/36

19.7 623.3 (+2) 30 229.1 (b2)/674.4 (y6)/545.4 (y5)/432.3 (y4)/331.2 (y3) 30/30/30/30/30 36/36/36/36/36
13 DIDDLELTLAK 21,42
- 623.3 (+2) 136 201.1 (a2)/229.1 (b2)/1017.5 (y9)/459.2 (b4)/218.1 (y2) 31/25/27/25/25 14/16/48/26/16
14.8 613.6 (+3) 30 805.9 (y152+)/741.4 (y142+)/602.4 (y6)/501.3 (y5)/430.3 (y4) 30/30/30/30/30 36/36/36/36/36
14 DLEEATLQHEATAAALR
9.37 613.6 (+3) 86 201.1 (a2)/229.1 (b2)/805.9 (y152+)/358.2 (b3)/501.3 (y5) 33/27/23/23/30 10/12/38/26/36
11.0 432.9 (+3) 30 359.2 (y3) b/589.3 (y5) b/474.3 (y4) b/702.4 (y6) b/839.4 (y7) b 30/30/30/30/30 36/36/36/36/36
15 DTQIHLDDALR
- 432.9 (+3) 61 476.8 (y82+)/217.1 (b2)/420.2(y72+)/589.3 (y5)/359.2 (y3) 15/15/21/23/27 18/10/24/26/18
b
16 EQDTSAHLER 1.5 593.3 (+3) 30 417.2 (y3)/554.3 (y4)/625.3 (y5)/813.4 (y7)/632.3 (b6) 30/30/30/30/30 36/36/36/36/36

13.5 744.9 (+2) 30 243.1 (b2)/215.1 (a2)/504.2 (y4)/375.2 (y3)/875.4 (y7) 30/30/30/30/30 36/36/36/36/36
17 IEELEEEIEAER *
- 744.9 (+2) 166 243.1 (b2)/372.2 (b3)/375.2 (y3)/504.2 (y4)/875.4 (y7) 35/33/31/35/33 12/20/20/24/50

18 IQHELEEAEER 2.0 691.8 (+2) 30 571.3 (y92+)/875.4 (y7)/379.2 (b3)/304.2 (y2)/504.2 (y4) 30/30/30/30/30 36/36/36/36/36

12.1 542.8 (+2) 30 242.1 (b2)/843.5 (y7)/730.4 (y6)/601.4 (y5)/355.2 (b3) 30/30/30/30/30 36/36/36/36/36
19 IQLELNQVK *
- 542.8 (+2) 111 242.1 (b2)/843.5 (y7)/355.2 (b3)/488.3 (y4)/730.4 (y6) 25/23/23/33/30 16/52/18/24/36
9.5 565.3 (+2) 30 229.1 (b2)/772.5 (y7)/701.4 (y6)/331.2 (y3)/901.5 (y8) 30/30/30/30/30 36/36/36/36/36
20 LDEAEQLALK *
7.54 565.3 (+2) 116 229.1 (b2)/201.1 (a2)/358.2 (b3)/901.5 (y8)/331.2 (y3) 27/33/25/21/27 12/14/18/48/16
28.8 658.3 (+3) 30 461.2 (y5)/702.4 (y7)/261.2 (y3)/390.2 (y4)/726.4 (b6) 30/30/30/30/30 36/36/36/36/36
21 NLQQEISDLTEQIAEGGK 42
- 658.3 (+3) 106 261.2 (y3)/390.2 (y4)/461.2 (y5)/200.1 (a2)/574.3 (y6) 19/23/27/37/27 12/20/22/10/30

13
Table S7: Continued.

22 TEELEEAK - - - a - -

11.7 570.7 (+3) 30 228.1 (b2)/362.2 (y3)/475.3 (y4)/741.9 (y13 2+)/675.4 (y6) 30/30/30/30/30 36/36/36/36/36
23 VQLLHTQNTSLINTK
- 570.7 (+3) 71 228.1 (b2)/362.2 (y3)/248.2 (y2)/741.9 (y132+)/475.3 (y4) 25/33/39/30/31 24/18/22/36/20

24 YETDAIQR 1.7 498.2 (+2) 30 703.4 (y6)/265.1 (a2)/487.3 (y4)/293.1 (b2)/602.3 (y5) 30/30/30/30/30 36/36/36/36/36

11.5 432.9 (+3) 30 359.2 (y3)/589.3 (y5)/474.3 (y4)/702.4 (y6)/839.4 (y7) 30/30/30/30/30 36/36/36/36/36
25 DTQLHLDDALR
- 432.9 (+3) 26 540.8 (y92+)/476.8 (y82+)/217.1 (b2)/420.2 (y72+)/359.2 (y3) 13/15/17/21/27 20/20/12/20/3
3.4 484.3 (+2) 30 209.1 (a2)/237.1 (b2)/731.4 (y5)/333.2 (y3)/446.3 (y4) 30/30/30/30/30 36/36/36/36/36
26 HVLATLGEK
3.08 484.3 (+2) 116 209.1 (a2)/237.1 (b2)/731.4 (y7)/333.2 (y3)/258.1 (y2-H2O) 31/25/23/33/37 12/16/38/16/14
9.1 596.8 (+2) 30 645.3 (y5)/532.2 (y4)/744.4 (y6)/417.2 (y3)/845.4 (y7) 30/30/30/30/30 36/36/36/36/36
27 EAFTVIDQNR
7.44 596.8 (+2) 91 645.3 (y5)/417.2 (y3)/532.2 (y4)/201.1 (b2)/744.4 (y6) 27/39/21/27/27 40/20/24/22/48
29.0 994.5 (+2) 30 712.3 (y6)/972.5 (y8)/611.3 (y5)/333.2 (y3)/480.2 (y4) 30/30/30/30/30 36/36/36/36/36
28 EASGPINFTVFLTMFGEK
- 663.3 (+3) 111 712.3 (y6)/480.2 (y4)/972.5 (y8)/333.2 (y3)/825.4 (y7) 27/39/25/39/27 34/24/54/16/50

29 ALESPERPFLAILGGAK 24.8 590.3 (+3) 30 792.9 (y152+)/728.4 (y142+)/684.9 (y132+)/332.2 (y4)/275.2 (y3) 30/30/30/30/30 36/36/36/36/36

25.5 879.9 (+2) 30 770.9 (b142+)/320.2 (y3)/219.1 (y2)/680.3 (y6)/566.2 (y5) 30/30/30/30/30 36/36/36/36/36
30 AISEELDHALNDMTSI
- 879.9 (+2) 66 219.1 (y2)/320.2 (y3)/680.3 (y6)/302.2 (y3-H2O)/201.1 (y2-H2O) 37/33/30/35/41 20/22/36/16/10
26.5 879.9 (+2) 30 320.2 (y3)/219.1 (y2)/566.2 (y5)/770.9 (b14 2+)/680.3 (y6) b 30/30/30/30/30 36/36/36/36/36
31 AISEELDHALNDMTSL
- 879.9 (+2) 171 320.2 (y3)/770.9 (b142+)/219.1 (y2)/1079.5 (b10)/680.3 (y6) 33/27/49/37/30 16/40/12/18/36
13.3 622.3 (+2) 30 242.1 (b2)/790.4 (y6)/355.2 (b3)/889.4 (y7)/661.3 (y5) 30/30/30/30/30 36/36/36/36/36
32 IQLVEEELDR 21,42
9.21 622.3 (+2) 121 242.1 (b2)/1002.5 (y8)/355.2 (b3)/790.4 (y6)/889.4 (y7) 29/25/27/29/29 14/46/20/42/54
14.4 566.3 (+2) 30 630.4 (y5)/758.4 (y6)/871.5 (y7)/501.3 (y4)/388.3 (y3) 30/30/30/30/30 36/36/36/36/36
33 MEIQEIQLK *
9.61 566.3 (+2) 116 233.1 (a2)/371.2 (y3-NH3)/871.5 (y7)/758.4 (y6)/260.2 (y2) 31/39/25/23/35 12/30/52/24/18
16.7 769.9 (+2) 30 346.2 (y3)/509.3 (y4)/995.5 (y8)/866.4 (y7)/622.4 (y5) 30/30/30/30/30 36/36/36/36/36
34 SIDDLEDELYAQK
10.50 769.9 (+2) 156 201.1 (b2)/346.2 (y3)/509.3 (y4)/258.1 (y2-NH3)/866.4 (y7) 37/39/37/53/37 26/24/38/24/26

35 SLEAQAEK 1.6 438.2 (+2) 30 201.1 (b2)/675.3 (y6)/347.2 (y3)/546.3 (y5)/475.3 (y4) 30/30/30/30/30 36/36/36/36/36

14
Table S7: Continued.

12.4 575.3 (+2) 30 261.1 (b2) b/648.3 (y5) b/776.4 (y6) b/889.5 (y7) b/519.3 (y4) b 30/30/30/30/30 36/36/36/36/36
36 MEIQEMQLK
- 575.3 (+2) 71 261.1 (b2)/233.1 (a2)/371.2 (y3-NH3)/889.5 (y7)/519.3 (y4) 25/29/41/25/27 28/16/18/38/30

37 AISEELDNALNDITSL 28.9 859.4 (+2) 30 219.1 (y2)/320.2 (y3)/530.2 (b5)/662.3 (y6)/775.4 (y7) 30/30/30/30/30 36/36/36/36/36

13.0 575.3 (+2) 30 261.1 (b2)/648.3 (y5)/776.4 (y6)/889.5 (y7)/519.3 (y4) 30/30/30/30/30 36/36/36/36/36
38 MELQEMQLK
- 575.3 (+2) 116 261.1 (b2)/233.1 (a2)/889.5 (y7)/371.2 (y3-NH3)/374.2 (b3) 27/33/23/39/25 24/24/48/20/18
22.8 513.8 (+2) 30 229.1 (b2)/798.4 (y6)/536.3 (y4)/651.3 (y5)/913.4 (y7) 30/30/30/30/30 36/36/36/36/36
39 IDFDEFLK
13.10 513.8 (+2) 111 229.1 (b2)/798.4 (y6)/201.1 (a2)/913.4 (y7)/260.2 (y2) 21/21/23/23/33 4/50/10/50/22
28.7 725.3 (+2) 30 554.3 (y4)/425.2 (y3)/814.4 (y6)/667.3 (y5)/943.5 (y7) 30/30/30/30/30 36/36/36/36/36
40 IDYDEFLEFMK
- 725.3 (+2) 201 229.1 (b2)/201.1 (a2)/554.3 (y4)/425.2 (y3)/814.4 (y6) 31/31/27/27/30 14/20/26/26/36

41 IMLQATGETITEDDIEELMK 27.6 760.7 (+3) 30 649.3 (y5)/611.8 (y102+)/992.5 (y8)/520.3 (y4)/278.2 (y2) 30/30/30/30/30 36/36/36/36/36

42 NADGYIDLEELK 17.3 690.3 (+2) 30 746.4 (y6)/859.5 (y7)/521.2 (b5)/389.2 (y3)/260.2 (y2) 30/30/30/30/30 36/36/36/36/36

43 ILLAELEQLK 23.2 585.4 (+2) 30 227.2 (b2)/830.5 (y7)/943.5 (y8)/759,4 (y6)/630.4 (y5) 30/30/30/30/30 36/36/36/36/36

* Observable L/I-isomers of entries 17, 19, 20 and 33 (tR for non-optimized LC; presumed specificity):

IEELEEELEAER (12.7 min; mammals and poultry), LQLELNQVK (12.7 min; poultry), LDEAEQIALK (8.9 min; mammals), MELQEIQLK (15.1 min; mammals).

15
Table S8: Presence of 62 peptides suitable to indicate meat in 19 food-relevant mammalian and poultry species (■) and their sequence homologies in green plants

(bold = Peptide sequence; light = L/I-isomers) according to the NCBI database. The 43 peptides preselected as meat markers are marked gray. The NCBI accessions

for the target proteins are representatives from pig (Sus scrofa), with mismatched amino acids marked bold. a = Peptide contains cysteine; b = > 20 amino acids;

c = Not originating from a myofibrillar or sarcoplasmic protein; d = Non-tryptic homology; e = Putative tryptic homology.

Mammals Poultry

A. platyrhynchos
Tryptic homologies
Target protein

M. gallopavo
O. cuniculus

N. meleagris
A. cygnoides
C. capreolus

C. moschata

P. colchicus
Peptide sequence in food-relevant

C. japonica
E. caballus

S. camelus
C. elaphus
(Sus scrofa)

B. taurus

C. hircus

G. gallus
S. scrofa
plants (selected)

A. anser
O. aries

C. livia
Arachis hypogea,
AGFAGDDAPR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Capsicum annuum,
Zea mays (and others)
Capsicum annuum,
Malus domestica,
AVFPSIVGRPR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Actin α, Solanum tuberosum
skeletal muscle (and others)
(XP_020927589.1) Hordeum vulgare,
DSYVGDEAQSK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Zea mays
Capsicum annuum,
Malus domestica,
IWHHTFYNELR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Solanum tuberosum
(and others)

16
Table S8: Continued.

Hordeum vulgare,
SYELPDGQVITIGNER Actin α, ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Cucurbita maxima,
skeletal muscle Zea mays (and others)
VAPEEHPTLLTEAPLNPK (XP_020927589.1) ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Zea mays
Adenylate kinase
YGYTHLSTGDLLR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(XP_003122225.3)
GNPTVEVDLHTAK Alpha-enolase ■ ■ ■ ■ ■ ■ ■
SGETEDTFIADLVVGLCTGQIK a,b (XP_020950937.1) ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Beta-enolase
IEEALGDK 42 ■ ■ ■ ■ ■ ■ ■
(NP_001037992.1)
GFSGLDGAK c Collagen α-1 ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
GPAGPQGPR c (XP_020922812.1) ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Prunus armeniaca,
LMVEMEK Creatine kinase, ■ ■ ■ ■ ■ ■ ■ ■
Prunus dulcis,
M-type Prunus mume,
(NP_001123421.1) Prunus persica
TDLNHENLK ■ ■ ■ ■ ■ ■ ■ ■ ■
Fructose-bisphosphate
GILAADESTGSIAK aldolase A ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(XP_020943654.1)
GAAQNIIPASTGAAK 42 ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Glyceraldehyde
VIISAPSADAPMFVMGVNHEK b 3-phosphat ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
dehydrogenase Prunus armeniaca,
VPTPNVSVVDLTCR a (XP_020946773.1) ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Solanum tuberosum,
Zea mays (and others)
DFNMPLTISR LIM domain-binding ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
protein 3
DSALSTHKPIEVK (XP_005657482.2) ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
LQDAEEHVEAVNAK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Myosin I
LQNEVEDLMIDVER 42 ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(NP_001098421.1)
SLSTELFK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Carya illinoinensis

17
Table S8: Continued.

ADIAESQVNK ■ ■ ■ ■ ■■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
AEDEEEINAELTAK ■ ■ ■ ■ ■■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
DIDDLELTLAK 21,42 ■ ■ ■ ■ ■■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
DLEEATLQHEATAAALR ■ ■ ■ ■ ■■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
DTQIHLDDALR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
ELEEISER ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Cicer arietinum
EQDTSAHLER Myosin II ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
IEELEEEIEAER (NP_999301.1) ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
IQHELEEAEER ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
IQLELNQVK ■ ■ ■ ■ ■ ■ ■ ■
LDEAEQLALK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
NLQQEISDLTEQIAEGGK 42 ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
TEELEEAK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
VQLLHTQNTSLINTK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
YETDAIQR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Myosin VIII
DTQLHLDDALR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(XP_020921928.1)
Myosin,
HVLATLGEK light chain 1/3 ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(NP_999539.2)
EAFTVIDQNR Myosin, ■ ■ ■ ■ ■ ■ ■ ■
regulatory light chain 2
EASGPINFTVFLTMFGEK (NP_001006592.1) ■ ■ ■ ■ ■ ■
Phosphoglycerate
ALESPERPFLAILGGAK kinase ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(NP_001093402.1)
Triosephosphate Brassica napus,
Brassica oleracea,
VVLAYEPVWAIGTGK isomerase ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Sesamum indicum,
(NP_001032228.1) Sinapis alba

18
Table S8: Continued.

AISEELDHALNDMTSI ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
AISEELDHALNDMTSL ■ ■ ■ ■ ■ ■
CAELEEELK a ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Tropomyosin α-1
IQLVEEELDR 21,42 ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(NP_001090952.1)
MEIQEIQLK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
SIDDLEDELYAQK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
SLEAQAEK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Tropomyosin α-4
MEIQEMQLK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(NP_999500.1)
AISEELDNALNDITSL ■ ■ ■ ■ ■ ■ ■
Tropomyosin β
IVTNNLK ■ ■ ■ ■ ■ ■ ■ ■ Lactuca sativa
(XP_020936634.1)
MELQEMQLK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
Capsicum annuum,
Solanum tuberosum,
ELGTVMR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Triticum aestivum
Troponin C, (and others)
fast skeletal muscle Cajanus cajan d,
(NP_001001862.1) Colocasia esculenta d,
IDFDEFLK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ Citrus sinensis d,
Citrus unshiu d,
Helianthus annuus e
IDYDEFLEFMK ■ ■ ■ ■ ■ ■ ■
AAFDIFVLGAEDGCISTK a Troponin C, ■ ■ ■ ■ ■ ■ ■ ■
slow skeletal muscle
IMLQATGETITEDDIEELMK (NP_001123715.1) ■ ■ ■ ■ ■ ■ ■ ■
NADGYIDLEELK ■ ■ ■ ■ ■ ■ ■ ■
Vimentin
ILLAELEQLK ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■
(XP_005668163.1)

19
Table S9: Peak areas (least intense fragment ion, Table S7) of the 43 peptides preselected as meat markers in seven food-important types of meat (n = 1, each)

applying non-optimized MRM. MEAN = Mean peak area in all seven samples; MEAN [%] = Percentage of a given peptide’s MEAN compared to the MEAN (100%)

of the most intense peptide (IQLVEELDR). Peptides (or their L/I-isomers) exhibiting ≥ 2% MEAN [%] are marked gray.

No. Peptide sequence Pork Beef Lamb Chicken Turkey Duck Goose MEAN MEAN [%]
1 YGYTHLSTGDLLR 27549 17532 22256 93847 20570 21131 37895 34397 0.2
2 GNPTVEVDLHTAK 49613 27421 31763 153282 85816 30382 43115 60199 0.3
3 IEEALGDK 42 2248 4437 14398 20043 8783 5075 6192 8740 0.0
4 TDLNHENLK 40903 101189 227323 89039 25455 32426 60720 82436 0.4
5 GILAADESTGSIAK 1210431 1508208 917661 10007638 2506513 2368132 3914194 3204682 14.4
6 GAAQNIIPASTGAAK 42 528994 1118798 365306 964472 296387 175614 290862 534348 2.4
7 DFNMPLTISR 522628 923639 441345 716508 594068 359236 427066 569213 2.6
8 DSALSTHKPIEVK 29681 51027 8240 14378 42583 27375 41029 30616 0.1
9 LQDAEEHVEAVNAK 4368 0 19286 3692 5527 4599 3519 5856 0.0
10 LQNEVEDLMIDVER 42 268879 51183 165367 182546 95353 264715 366658 199243 0.9
11 ADIAESQVNK 17741 9153 118959 9994 18026 13266 16521 29094 0.1
12 AEDEEEINAELTAK 15101 73358 86254 10457 11420 12440 14768 31971 0.1
13 DIDDLELTLAK 21,42 748030 671533 399694 405535 416619 509572 673109 546299 2.4
14 DLEEATLQHEATAAALR 974078 1067337 750087 768385 879553 537311 1103319 868581 3.9
15 DTQIHLDDALR Undetectable by MRM; see Table S7.
16 EQDTSAHLER 9408 3066 83176 7774 6140 9327 6892 17969 0.1
17 IEELEEEIEAER 24400 93289 190624 -* - - - 102771 0.5
18 IQHELEEAEER 24757 8634 85063 16892 26830 17949 17431 28222 0.1
19 IQLELNQVK 1747249 645540 603199 -* - - - 998662 4.5
20 LDEAEQLALK 5264473 1962846 1784471 4082802 4306587 3838132 4673724 3701862 16.6
21 NLQQEISDLTEQIAEGGK 42 771512 193739 333607 179107 937065 677553 781708 553470 2.5

20
Table S9: Continued.

22 TEELEEAK Undetectable by HPLC-LR-MS/MS; see Table S7 and Section 3.4.

23 VQLLHTQNTSLINTK 2509159 1164917 814720 673840 1591601 1126365 1431789 1330342 6.0
24 YETDAIQR 114417 79789 115004 90834 238701 79417 180292 128351 0.6
25 DTQLHLDDALR 104807 88480 37561 251381 309808 218188 262583 181830 0.8
26 HVLATLGEK 9608381 5553875 3425039 12942127 8538317 1279684 2129510 6210990 27.8
27 EAFTVIDQNR 14023076 11284372 10346172 19134654 9367005 6962003 7278104 11199341 50.2
28 EASGPINFTVFLTMFGEK 1554490 63788 1268881 2103114 1610009 813372 1024227 1205411 5.4
29 ALESPERPFLAILGGAK 41003 66723 15720 212044 34226 42098 77942 69965 0.3
30 AISEELDHALNDMTSI 1617888 1511102 1383438 2836935 1798617 769464 755376 1524689 6.8
31 AISEELDHALNDMTSL 76101 45207 63291 50047 79124 52190 60117 60868 0.3 **
32 IQLVEEELDR 21,42 28315915 29046983 23399675 19472826 17518909 18688056 19753773 22313734 100.0
33 MEIQEIQLK 11882314 5353440 8180972 13557435 9256656 7251620 8715130 9171081 41.1
34 SIDDLEDELYAQK 8068522 3484292 6816984 8581878 6607211 7320323 7739709 6945560 31.1
35 SLEAQAEK 3222 5593 76412 3729 5940 7652 9574 16017 0.1
36 MEIQEMQLK Undetectable by MRM **; see Table S7 and Section 3.5.
37 AISEELDNALNDITSL 657394 744842 396280 255918 218293 57912 46023 339523 1.5
38 MELQEMQLK 5593861 8040497 2291589 397788 1884062 371383 470229 2721344 12.2
39 IDFDEFLK 9264867 5586057 7457720 6285986 6654363 7419418 6569407 7033974 31.5
40 IDYDEFLEFMK 345550 1324170 407246 278794 328701 232061 394855 473054 2.1
41 IMLQATGETITEDDIEELMK 226047 951041 261088 232070 211386 149880 260586 327442 1.5
42 NADGYIDLEELK 106325 517223 147500 113593 69706 68433 113425 162315 0.7
43 ILLAELEQLK 142319 232735 237772 860952 446658 713604 329797 423405 1.9

* Entries 17 und 19 are presumed mammalian specific peptides; see Table S7, Section 3.4., and Section 3.5.

** The L/I-isomers of entries 31 and 36 – entries 30 and 38 – exhibited ≥ 2% MEAN [%].

21
Table S10: Peak areas (qualifier 2, Table S7) of the 18 potential marker peptide candidates in full-preserve vegan bratwurst-style sausages spiked with 5 %, w/w pork

or 5 %, w/w chicken meat (n = 3, each) applying (fully) optimized MRM. MEAN = Mean Peak area in all 18 samples; MEAN [%] = Percentage of a given peptide’s

MEAN compared to the MEAN (100%) of the most intense peptide (IQLVEELDR). Peptides exhibiting ≥ 10% MEAN [%] are marked gray.

Pork Chicken Total

Peptide sequence
Peak areas MEAN MEAN [%] Peak areas MEAN MEAN [%] MEAN MEAN [%]
AISEELDHALNDMTSI 18702 22351 16228 19094 1.0 57087 52861 60835 56928 2.7 38011 1.9
DFNMPLTISR 183048 215011 166230 188096 9.4 248363 226861 252749 242658 11.7 215377 10.6
DIDDLELTLAK 21,42 100919 111511 92558 101663 5.1 133050 114674 110989 119571 5.7 110617 5.4
DLEEATLQHEATAAALR 211344 221792 178662 203933 10.2 565370 457481 443159 488670 23.5 346301 17.0
EAFTVIDQNR 179667 178752 162329 173582 8.7 280800 233540 266317 260219 12.5 216901 10.7
EASGPINFTVFLTMFGEK 3336 3863 5403 4201 0.2 3221 2781 2205 2736 0.1 3468 0.2
GAAQNIIPASTGAAK 42 57213 55730 53461 55468 2.8 147567 107190 112569 122442 5.9 88955 4.4
GILAADESTGSIAK 178675 174697 156153 169842 8.5 440796 357286 346260 381447 18.3 275645 13.5
HVLATLGEK 380308 317745 350893 349649 17.6 1079950 799913 955685 945183 45.4 647416 31.8
IDFDEFLK 489839 525878 454316 490011 24.6 571517 527860 517570 538982 25.9 514497 25.3
IDYDEFLEFMK 10342 13372 10970 11561 0.6 7691 6518 6614 6941 0.3 9251 0.5
IQLVEEELDR 21,42 2071408 2114578 1786634 1990873 100.0 2332720 1967195 1944900 2081605 100.0 2036239 100.0
LDEAEQLALK 306879 332411 294206 311165 15.6 575232 469672 476619 507174 24.4 409170 20.1
MEIQEIQLK 282960 281064 216987 260337 13.1 732332 646482 755210 711341 34.2 485839 23.9
MELQEMQLK 151623 207909 147948 169160 8.5 10628 10702 10912 10747 0.5 89954 4.4
NLQQEISDLTEQIAEGGK 42 108405 109086 89822 102437 5.1 67995 59458 56286 61246 2.9 81842 4.0
SIDDLEDELYAQK 121148 127482 110936 119855 6.0 384327 344834 332536 353899 17.0 236877 11.6
VQLLHTQNTSLINTK 98432 99754 89651 95946 4.8 66068 55571 55104 58914 2.8 77430 3.8

22
Figure S1: Chromatograms derived from the pooled samples “Common Cereals” (A), “Beans” (B), and

“Other Legumes” (C). Low signals (see y-axis for scale) of the marker peptide candidate

GILAADESTGSIAK were detectable all pooled samples except B. black = quantifier; blue = qualifier 1;

red = qualifier 2.

23
Table S11: S/N (qualifier 2, Table S7) of the 9 remaining marker peptide candidates in spray-dried milk,

egg, casein, whey, albumen, and yolk (Table S4), and in 10-fold diluted samples of beef and chicken meat

(Table S1) (n = 3, each), applying fully optimized MRM. The listed values already account for the dilution.

gray = S/N ≥ 10; dark gray = S/N ≥ 1,000.

Milk Milk Milk Egg Egg Egg 3

Peptide sequence
1 2 3 1 2
DFNMPLTISR - - - - - -
DLEEATLQHEATAAALR - - - - - -
EAFTVIDQNR - - - - - 5
HVLATLGEK 4 5 4 5 4 9
IDFDEFLK - - - - - 10
IQLVEEELDR 21,42 - 5 - - - 9
LDEAEQLALK - - - - - -
MEIQEIQLK - - - - - -
SIDDLEDELYAQK - - - - - -
Casein Casein Casein Whey Whey Whey
Peptide sequence
1 2 3* 1 2 3*
DFNMPLTISR - - - - - -
DLEEATLQHEATAAALR 40 4 - - - -
EAFTVIDQNR 24 15 - - - -
HVLATLGEK 58 17 - - - -
IDFDEFLK 9 - - - - -
IQLVEEELDR 21,42 15 6 - 26 12 8
LDEAEQLALK 40 38 - - - -
MEIQEIQLK - - - - - -
SIDDLEDELYAQK - - - - - -
Albumen Albumen Albumen Yolk Yolk Yolk
Peptide sequence 1 2 3* 1 2 3*
DFNMPLTISR - - - - - -
DLEEATLQHEATAAALR - - - 23 25 -
EAFTVIDQNR - - - 37 21 -
HVLATLGEK - 10 - 28 24 -
IDFDEFLK - 7 - 70 - -
IQLVEEELDR 21,42 - 8 - 14 9 -
LDEAEQLALK - 19 - 27 46 -
MEIQEIQLK - - - - - -
SIDDLEDELYAQK - - - - - -

24
Table S11: Continued.

Beef Beef Beef Chicken Chicken Chicken

Peptide sequence
1 2 3 1 2 3
DFNMPLTISR 3196 2351 4987 3185 3412 3449
DLEEATLQHEATAAALR 4863 3533 3585 2677 3859 2723
EAFTVIDQNR 2538 2468 2856 2987 2841 3260
HVLATLGEK 4122 3542 3566 2490 4738 3148
IDFDEFLK 1460 1009 2043 1456 1367 1284
IQLVEEELDR 21,42 25374 18022 20413 38327 24971 27764
LDEAEQLALK 1516 1440 2978 3973 5278 4268
MEIQEIQLK 7265 17098 13125 33477 29156 25222
SIDDLEDELYAQK 3857 4368 4207 3971 2740 3166

* Sample materials derived from products intended for laboratory use, for which a higher purity

compared to products intended for human consumption was assumed.

25
Figure S2: Chromatogram derived from the commercial meat substitute “Vegetarian geschnetzeltes”

(milk content: 77% of the total weight). No peptide signals were detectable.

26
Table S12: Peak areas (quantifier, Table S7) of the 9 remaining marker peptide candidates in 10-fold diluted samples of 19 food-relevant types of mammalian and

poultry meat (three different animals, each) applying fully optimized MRM. The listed values already account for the dilution. SD = Standard deviation.

DLEEATLQHEATAAALR

SIDDLEDELYAQK
IQLVEEELDR 21,42

LDEAEQLALK
EAFTVIDQNR
DFNMPLTISR

HVLATLGEK

MEIQEIQLK
Type of Meat

IDFDEFLK
17685150 51603122 65101535 68316055 49400890 221278515 104039290 103905625 17979814
Pork
17060537 58450724 82039691 106134089 70622216 249307953 139967420 140330455 24630138
S. Scrofa
18001419 82427946 85189717 109295393 70962146 257568973 149459181 142488909 24572564
MEAN 17582369 64160597 77443648 94581846 63661751 242718480 131155297 128908330 22394172
SD [%] 2.2 20.6 11.4 19.7 15.8 6.4 14.9 13.7 13.9
16050385 64178492 37343940 63277113 34712141 162044449 54090060 58236040 9273865
Beef
17445848 68980667 67260509 97214316 65201391 235680019 76328243 95453911 16694809
B. taurus
21639113 74110557 57160131 88405162 45962617 269704441 56278127 78886136 14374285
MEAN 18378449 69089905 53921527 82965530 48625383 222476303 62232143 77525362 13447653
SD [%] 12.9 5.9 23.0 17.3 25.9 20.2 16.1 19.6 23.0

27
Table S12: Continued.

18367840 69578804 59777469 94076906 53296184 213284430 64703123 122131217 22435010

Lamb
19372135 57995615 50257176 86387428 40691622 201218708 48652108 64518353 13460611
O. aries
17155969 54772410 43847787 78669631 36671067 173021917 46613481 54739995 11986564
MEAN 18298648 60782276 51294144 86377988 43552958 195841685 53322904 80459855 15960728
SD [%] 5.0 10.5 12.8 7.3 16.3 8.6 15.2 37.0 28.9
19982666 91253322 42958214 74618167 43676102 235215559 69368604 72131870 12206510
Goat
20231711 63094210 48310222 62122236 36300327 214499197 51711317 71269298 11801408
C. hircus
17520766 45332824 39574659 47374535 26303745 175470613 39325027 56038194 9321830
MEAN 19245048 66560119 43614365 61371646 35426725 208395123 53468316 66479787 11109916
SD [%] 6.4 28.4 8.2 18.1 20.1 11.9 23.1 11.1 11.5
19635965 27824940 90490519 144671293 77221536 253682140 113282409 191519783 35656981
Rabbit
25628853 56380352 108355299 142522394 56991530 257953345 142405424 152097138 30799476
O. cuniculus
24489748 19914101 95566452 120477188 54494152 245644338 104412996 143752431 29861733
MEAN 23251522 34706464 98137423 135890292 62902406 252426608 120033610 162456451 32106063
SD [%] 11.2 45.1 7.7 8.0 16.2 2.0 13.5 12.8 7.9
20379739 69911613 40778701 60891038 46560453 206906913 61620315 72962136 12513809
Horse
21394522 91858046 60791466 71697325 40443454 282888097 74628144 84394334 14797994
E. caballus
21776441 75988648 82396610 95332288 55321816 255784046 98017524 99857677 19886885
MEAN 21183567 79252769 61322259 75973550 47441908 248526352 78088661 85738049 15732896
SD [%] 2.8 11.7 27.7 18.9 12.9 12.7 19.3 12.9 19.6
24878377 106981576 71006735 96092147 58041599 309295162 71022552 104698768 16985560
Red deer
21016694 86775650 73458804 92411052 49409798 243888757 76422986 103382901 17926836
C. elaphus
20463816 85969095 70861956 94433020 53037767 248340016 80090608 105308291 18597408
MEAN 22119629 93242107 71775832 94312073 53496388 267174645 75845382 104463320 17836601
SD [%] 8.9 10.4 1.7 1.6 6.6 11.2 4.9 0.8 3.7

28
Table S12: Continued.

14944405 43206032 63839716 70959507 48015171 177327853 40798344 115218337 21711205

Roe deer
18001940 72709885 68377924 90009141 48665803 207924286 63382467 110350151 20712269
C. capreolus
18780528 54327474 75592691 86605849 46415479 212086428 55324732 130141234 24970725
MEAN 17242291 56747797 69270110 82524832 47698818 199112856 53168514 118569907 22464733
SD [%] 9.6 21.4 7.0 10.1 2.0 7.8 17.6 7.1 8.1
22186684 60620401 71644280 141016122 57926228 228890696 114238910 171884090 33012512
Chicken
22899406 62581901 76020260 143876119 61715822 276661558 113695749 206204566 39999717
G. gallus
23361453 165457190 96366682 133735403 56786620 255897132 187225582 178233710 34257629
MEAN 22815848 96219831 81343741 139542548 58809557 253816462 138386747 185440789 35756619
SD [%] 2.1 50.9 13.2 3.1 3.6 7.7 25.0 8.0 8.5
23208679 69473874 73382143 134822268 63193205 228171568 131583657 141099956 27289564
Turkey
28314181 116225864 94014946 156236980 75869298 271542006 167962444 183685365 35769175
M. gallopavo
31862607 150474547 111788075 177758728 79005931 296935429 187451062 195038744 38340618
MEAN 27795156 112058095 93061721 156272659 72689478 265549668 162332388 173274688 33799786
SD [%] 12.8 29.6 16.9 11.2 9.4 10.7 14.3 13.4 14.0
Duck 16575504 39991484 52134882 80140418 47800862 178993912 103071129 120504781 22539445
A. platyrhynchos 15560383 30117332 47463767 77908156 43151829 168501958 91404336 125336310 24453637
C. moschata 22262415 69130892 52592960 94716693 40004542 173033006 113480648 124108513 22337289
MEAN 18132767 46413236 50730536 84255089 43652411 173509625 102652038 123316535 23110124
SD [%] 16.3 35.7 4.6 8.8 7.3 2.5 8.8 1.7 4.1
Goose 18966464 75349115 54452455 74384950 46742444 195750055 111505963 124660148 23841923
A. anser 17675563 47231781 42817985 64530378 34543749 193520904 79354355 134048575 27135123
A. cygnoides 16295336 49018680 43698575 59704087 38274337 189544049 80633281 132026130 25957722
MEAN 17645788 57199859 46989672 66206472 39853510 192938336 90497866 130244951 25644923
SD [%] 6.2 22.5 11.3 9.2 12.8 1.3 16.4 3.1 5.3

29
Table S12: Continued.

15640300 86592659 62620989 109172282 47344513 198132414 138331677 153324082 28119077

Pheasant
20618698 86084258 83031287 96911705 33520308 246432405 127110447 153003770 30074250
P. colchicus
15103453 115284841 83637615 103149237 18434963 195285057 144288475 149005537 31198851
MEAN 17120817 95987253 76429964 103077741 33099928 213283292 136576866 151777796 29797393
SD [%] 14.5 14.2 12.8 4.9 35.7 11.0 5.2 1.3 4.3
12291563 78964944 49409600 77432263 28800279 164984244 114764322 125124495 23845599
Quail
13551136 61920742 36937994 63841961 26373677 149857863 110840597 104180291 21504663
C. japonica
17096911 111137020 60563108 87133666 28205671 164390394 140708138 121642983 24306919
MEAN 14313203 84007569 48970234 76135963 27793209 159744167 122104352 116982590 23219060
SD [%] 14.2 24.3 19.7 12.5 3.7 4.4 10.9 7.8 5.3
18064196 74601665 71814498 116257813 48603516 222743462 118943387 179808770 33944902
Guinea fowl
16096054 45585808 51239137 95764261 39597606 157860490 97517212 119966142 26658544
N. meleagris
20185221 143379558 91473008 122953767 39678656 199538383 145604982 152377318 30342412
MEAN 18115157 87855677 71508881 111658614 42626593 193380778 120688527 150717410 30315286
SD [%] 9.2 46.7 23.0 10.4 9.9 13.9 16.3 16.2 9.8
25919701 46264130 72682932 90659050 69513934 249568613 117804978 107394575 19825538
Ostrich
24366203 54065878 61200367 73825639 56256895 232364577 98665092 92132827 16919310
S. camelus
4468225 3286675 9964597 14206711 5247771 38824656 11725060 10006850 2499301
MEAN 18251376 34538894 47949299 59563800 43672867 173585949 76065043 69844751 13081383
SD [%] 53.5 64.6 56.9 55.1 63.4 55.0 60.7 61.2 53.5
20420844 83138280 70448792 92665828 35201540 227666328 118002226 168690560 33239631
Pigeon
22820539 126586918 64119375 77406651 32174463 165835617 127663464 116582439 21599687
C. livia
20596063 116491670 64344640 77115274 29098446 156539889 116398179 105943168 20922378
MEAN 21279149 108738956 66304269 82395918 32158150 183347278 120687956 130405389 25253899
SD [%] 5.1 17.1 4.4 8.8 7.7 17.2 4.1 21.0 5.1

30
Table S12: Continued.

Average MEAN 19574752 73385965 65298096 93712151 46891885 214460447 99841565 120976821 23001837

Average SD [%] 15.6 31.5 24.6 28.7 25.3 15.7 33.6 29.3 15.6
MEAN (Pork, Beef,
Chicken, Turkey) 21642955 85382107 76442659 118340646 60946542 246140228 123526644 141287292 26349558
SD [%] (Pork, Beef,
Chicken, Turkey) 18.8 23.0 18.6 25.7 14.2 6.4 30.1 30.0 34.3

31
Figure S3: Chromatograms derived from the 10-fold diluted samples of beef (A) and turkey (B) meat.

The signals of a given peptide detected in A and B were of comparable intensities (see y-axis for scale).

a = HVLATLGEK; b = EAFTVIDQNR; c = LDEAEQLALK; d = IQLVEEELDR; e =

DLEEATLQHEATAAALR; f = MEIQEIQLK; g = SIDDLEDELYAQK; h = DFNMPLTISR;

i = IDFDEFLK; * = MELQEIQLK (tR = 9.88 min), isomer of MEIQEIQLK, presumably indicating

mammalian meat; ** = putative, but uncharacterized isomers of LDEAEQLAK and IQLVEEELDR;

black = quantifier; blue = qualifier 1; red = qualifier 2.

32
Table S13: Peak areas (quantifier, see Table S7) of the 9 marker peptides in uncooked (upper rows) and canned (lower rows) vegan bratwurst-style sausage fillings

spiked with 5 %, w/w pork or 5 %, w/w chicken meat (n = 3, each) applying fully optimized MRM. MEAN = Mean peak area in all samples (canned or uncooked);

MEAN [%] = Percentage of a given peptide’s MEAN in canned sausage fillings compared to its MEAN in uncooked sausage fillings.

Pork (average MEAN [%] = -30.8%) Chicken (average MEAN [%] = -22.3%) Total (-25.7%)
Peptide sequence
Peak area MEAN MEAN [%] Peak area MEAN MEAN [%] MEAN MEAN [%]
219395 247412 261182 242663 235399 267435 248832 250555 246609
DFNMPLTISR -23.1 -4.4 -13.6
177379 184703 198003 186695 233540 254703 230244 239496 213095
976041 865435 848281 896586 1397760 1446100 1157245 1333702 1115144
DLEEATLQHEATAAALR +32.4 +57.9 +47.6
1058587 1151657 1350048 1186764 1852282 2270821 2195240 2106114 1646439
848895 848478 950017 882464 1241635 1234906 1290194 1255578 1069021
EAFTVIDQNR -59.5 -63.2 -61.6
350877 342716 378885 357493 429701 484014 474088 462601 410047
1093475 1136988 1389078 1206514 2025581 1641576 2234274 1967144 1586829
HVLATLGEK -52.8 -31.7 -39.7
532998 580720 595937 569885 1362795 1335738 1329741 1342758 956321
1035802 997036 933181 988673 972175 1009587 911755 964506 976589
IDFDEFLK -5.8 -9.0 -7.4
867038 938838 988731 931535 819794 949133 863617 877515 904525
6579064 6486164 6782976 6616068 5361972 5475085 5294733 5377264 5996666
IQLVEEELDR 21,42 -42.5 -40.4 -41.5
3640629 3749523 4029906 3806686 2890350 3491757 3228262 3203456 3505071
1740165 1693345 1646218 1693243 2022572 2138819 1826018 1995803 1844523
LDEAEQLALK -24.5 -24.5 -24.5
1146505 1284459 1402047 1277670 1304723 1655093 1561996 1507271 1392470
874719 1013009 1096469 994732 1932096 2227166 2166590 2108617 1551675
MEIQEIQLK -41.3 -37.0 -38.4
542511 577556 631375 583814 1226371 1459626 1297139 1327712 955763
314269 334589 349508 332789 692459 731112 723923 715831 524310
SIDDLEDELYAQK -60.3 -48.7 -52.4
119994 132129 144177 132100 346885 383648 370139 366891 249495

33