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This procedure consists of a milligram\u2010scale alkaline hydrolysis followed by an absolute quantification step using a multiple reaction monitoring tandem mass spectrometric (LC\u2010MS\/MS) detection method. The approach facilitates the analysis of a few hundred samples per week by using a 96\u2010well plate extraction setup. Importantly, the method uses only \u223c4 mg of tissue per sample and uses the common alkaline hydrolysis protocol, followed by water extraction that includes <jats:sc>L<\/jats:sc>\u2010Trp\u2010d5 as an internal standard to enable the quantification of the absolute level of the bound Trp with high precision, accuracy, and reproducibility. The protocol described herein has been optimized for seed samples for <jats:italic>Arabidopsis thaliana<\/jats:italic>, <jats:italic>Glycine max<\/jats:italic>, and <jats:italic>Zea mays<\/jats:italic> but could be applied to other plant tissues. \u00a9 2023 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol<\/jats:bold>: Analysis of protein\u2010bound tryptophan from seeds<\/jats:p>","DOI":"10.1002\/cpz1.862","type":"journal-article","created":{"date-parts":[[2023,8,4]],"date-time":"2023-08-04T18:43:50Z","timestamp":1691174630000},"source":"Crossref","is-referenced-by-count":3,"title":["A High\u2010Throughput Absolute Quantification of Protein\u2010Bound Tryptophan from Model and Crop Seeds"],"prefix":"10.1002","volume":"3","author":[{"given":"Huda","family":"Ansaf","sequence":"first","affiliation":[{"name":"Bond Life Sciences Center University of Missouri  Columbia Missouri"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abou","family":"Yobi","sequence":"additional","affiliation":[{"name":"Bond Life Sciences Center University of Missouri  Columbia Missouri"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruthie","family":"Angelovici","sequence":"additional","affiliation":[{"name":"Bond Life Sciences Center University of Missouri  Columbia Missouri"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,8,4]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.69.255"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021\u20109258(18)41947\u20103"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-030-77477-6_11"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13007\u2010018\u20100277\u20108"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1080\/15592324.2015.1096469"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021\u20109673(98)00721\u20103"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1177\/1178646918802282"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1515\/BC.2005.097"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00726\u2010018\u20102640\u20105"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021\u20109258(19)45285\u2010X"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021\u20109258(19)45285\u2010X"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2019.00797"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003\u20102697(80)90028\u20107"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1515\/bchm2.1984.365.2.1183"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac60139a005"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1155\/2016\/8952520"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/bip.21436"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcs.2007.06.006"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1093\/jexbot\/53.370.947"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.108.118091"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002170050197"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.3390\/molecules25020311"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00726\u2010009\u20100269\u20100"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpz1.861"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/cppb.20084"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2003.07.026"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj1350059"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.862","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,27]],"date-time":"2024-01-27T14:04:17Z","timestamp":1706364257000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.862"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8]]},"references-count":27,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2023,8]]}},"alternative-id":["10.1002\/cpz1.862"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.862","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,8]]},"article-number":"e862"}}