{"id":200,"date":"2022-05-16T17:29:01","date_gmt":"2022-05-16T17:29:01","guid":{"rendered":"http:\/\/chaletloree.com\/?page_id=200"},"modified":"2025-09-17T15:34:37","modified_gmt":"2025-09-17T15:34:37","slug":"biochemistry","status":"publish","type":"page","link":"http:\/\/chaletloree.com\/index.php\/biochemistry\/","title":{"rendered":"biochemistry"},"content":{"rendered":"\n<p><u>Sample preparation, quenching and metabolite extraction<\/u><\/p>\n\n<ul>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1_VA5gs_Bd5-03vflczkXAmHerAxgSJo-BY3aXNjRypc\/edit?usp=sharing\" title=\"\">Quenching and liquid\/liquid metabolite extraction from adherent cells<\/a><\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1HR5eCIAj6VP-56gctkC2uWDv4xb5Vx_FLOGJX91jRUM\/edit?usp=sharing\" title=\"\">Quenching and direct extraction of metabolites from adherent cells<\/a><\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1JUmkwt6WpB9Zny68mMXvNKzfdKbO4WP68K3HxViIyZY\/edit?usp=sharing\" title=\"\">Quenching of bacteria in solution (NaCl\/Ethanol)<\/a><\/li>\n\t<li>Quenching eukaryotic suspension cells<\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1H4lKfYqamBpf7tLSOwKfQcTWEDTut1qgIlVkDbtmUWA\/edit?usp=sharing\" title=\"\">Isolation of mitochondria from cell lines&nbsp;<\/a><\/li>\n<\/ul>\n\n<p><u>GC-MS usage<\/u><\/p>\n\n<ul>\n\t<li>general rules<\/li>\n\t<li>Preparing the machine and adding your samples<\/li>\n\t<li>Qualitative analysis<\/li>\n\t<li>Quantitative analysis<\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1aBU1UQ925Z6uJvf1hcMEMommkY98W6EuvPQ8c-OM6ro\/edit?usp=sharing\" title=\"\"><span style=\"font-size: 1rem;\">Derivatization with methoxyamine and MSTFA<\/span><\/a><\/li>\n\t<li>GC-MS analysis of polar metabolites<\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/10vmUvpYpWJOApkdAiFsTpLuNF3Gq63qJYNcFyQygZ1U\/edit?usp=sharing\" title=\"\">&#8220;Saponification&#8221; and methylation of fatty acids derived from lipids<\/a><\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1A2-Kiafrht63rc3USg4rmxe5MhjfZO0hHu5eRzu9Z9U\/edit?usp=sharing\" title=\"\">GC-MS analysis of fatty acid methylesters from cellular extracts<\/a><\/li>\n\t<li>GC-MS analysis of fatty acid methyl esters a la Joseph<\/li>\n<\/ul>\n\n<p><u>LC-MS usage<\/u><\/p>\n\n<ul>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1DLJ29-g3-fUN-skD8cuoGEGWtcxQBb8Oz95zCSA1ctQ\/edit?usp=sharing\" title=\"\">general rules for Agilent 6550 system<\/a> (french and english)<\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1zTBK-eqRbXf3EA8_p4MBEHtgbPhHQ2NSgAvDlpxXETA\/edit?usp=sharing\" title=\"\">Checklist before running your samples (french and english)<\/a><\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1-riZ1jY0rRAHPY0ec1jHfbfhNJnEg0HKi-tunMZOsfc\/edit?usp=sharing\" title=\"\">Preparing the machine and adding your samples<\/a><\/li>\n\t<li>Quantitative analysis<\/li>\n\t<li>Qualitative analysis<\/li>\n\t<li>Screening for unexpected changes of known or unknown metabolites<\/li>\n<\/ul>\n\n<p><u>HPLC<\/u><\/p>\n\n<ul>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1KRVglfrxjNFtGm9lU-AU1z4obHgm2KZjZFHOwHfUqUk\/edit?usp=sharing\" title=\"\">HPLC for nucleotides on SAX column<\/a><\/li>\n\t<li>Determination of H2S and thiosulfate by HPLC<\/li>\n<\/ul>\n\n<p><u>Determination of phosphate release<\/u><\/p>\n\n<ul>\n\t<li>Fiske-Subbarow<\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1ldrFPanMflVqBnDwPzIo1SxuXM8RDSqr_zzVEiF1ZIQ\/edit?usp=sharing\" title=\"\">Molybdate-Malachite Green method (Itaya)<\/a><\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1NhdpsE9DHWut_xikPnzZzgRZCTFEVC9lSht7VMdDYH0\/edit?usp=sharing\" title=\"\">Mild conditions (Saheki)<\/a><\/li>\n<\/ul>\n\n<p><u>Thin layer chromatography:<\/u><\/p>\n\n<ul>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1hXWIJGEWT1KHIhyq_Er93F-hfjKP2FbKEkqaYX109L8\/edit?usp=sharing\" title=\"\">Thin layer chromatography of lipids<\/a><\/li>\n\t<li><a href=\"https:\/\/docs.google.com\/document\/d\/1-QTujv2mbpQAbIfxdixk149LFAhsLeJdfpBlNzZb2tQ\/edit?usp=sharing\" title=\"\">Staining lipids with Primulin<\/a><\/li>\n<\/ul>\n\n<p><u><a href=\"index.php\/protocols\/biochemical-methods\/specific-metabolite-determinations\">Determination of metabolites and enzymatic activities<\/a><\/u><\/p>\n\n<p><u>Manipulation of metabolites<\/u><\/p>\n\n<ul>\n\t<li>Desalting metabolites by gel filtration<\/li>\n\t<li>Reduction of carbonyl groups with Sodium borohydride\/-deuteride<\/li>\n<\/ul>\n\n<p>&nbsp;<\/p>\n\n<p>&nbsp;<\/p>\n\n","protected":false},"excerpt":{"rendered":"<p>Sample preparation, quenching and metabolite extraction Quenching and liquid\/liquid metabolite extraction from adherent cells Quenching and direct extraction of metabolites from adherent cells Quenching of bacteria in solution (NaCl\/Ethanol) Quenching eukaryotic suspension cells Isolation of mitochondria from cell lines&nbsp; GC-MS usage general rules Preparing the machine and adding your samples Qualitative analysis Quantitative analysis Derivatization [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-200","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/200","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/comments?post=200"}],"version-history":[{"count":2,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/200\/revisions"}],"predecessor-version":[{"id":294,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/200\/revisions\/294"}],"wp:attachment":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/media?parent=200"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}