{"id":43562,"date":"2021-01-12T13:30:00","date_gmt":"2021-01-12T13:30:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/analisi-diretta-e-rapida-degli-amminoacidi-non-derivatizzati-nelle-formule-per-lattanti\/"},"modified":"2025-12-19T14:15:06","modified_gmt":"2025-12-19T14:15:06","slug":"analisi-diretta-e-rapida-degli-amminoacidi-non-derivatizzati-nelle-formule-per-lattanti","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/it\/articoli\/fffa3436-it\/analisi-diretta-e-rapida-degli-amminoacidi-non-derivatizzati-nelle-formule-per-lattanti","title":{"rendered":"Analisi diretta e rapida degli amminoacidi non derivatizzati nelle formule per lattanti"},"content":{"rendered":"\n<ul class=\"wp-block-list\">\n<li>Preparazione semplice e in un\u2019unica fase del campione per l\u2019analisi diretta degli amminoacidi non derivatizzati.<\/li>\n\n\n\n<li>Analisi simultanea degli amminoacidi non polari, polari, con carica positiva e con carica negativa in una breve corsa di soli 10 minuti.<\/li>\n\n\n\n<li>Un gradiente ben definito separa i contaminanti dagli analiti target.<\/li>\n<\/ul>\n\n\n\n<p>Per soddisfare le esigenze nutrizionali, \u00e8 necessario ricorrere a metodi analitici affidabili che determinino con accuratezza gli amminoacidi liberi nelle formule per lattanti. Una derivatizzazione precolonna seguita da un\u2019analisi LC a fase inversa \u00e8 un approccio tipico in cui la derivatizzazione \u00e8 necessaria a causa della mancanza di ritenzione cromatografica e scarsa sensibilit\u00e0 degli amminoacidi liberi. Tuttavia, si tratta di metodi che richiedono molto tempo e una procedura elaborata. Un\u2019altra tecnica comune \u00e8 l\u2019utilizzo di acidi perfluorurati come reagenti di coppia ionica per migliorare la ritenzione degli amminoacidi non derivatizzati su una colonna C18, ma questo metodo pu\u00f2 avere un impatto negativo sul sistema cromatografico e sullo spettrometro di massa.<\/p>\n\n\n\n<p>Nel metodo pi\u00f9 semplice qui illustrato, gli amminoacidi non derivatizzati possono essere misurati direttamente \u2013 dopo aver eseguito una preparazione del campione in un\u2019unica fase \u2013 utilizzando una colonna Raptor Polar X insieme a un detector MS\/MS. Le colonne Raptor Polar X presentano una fase ibrida (HILIC e a scambio ionico) che offre la ritenzione bilanciata necessaria per l\u2019analisi simultanea di un\u2019ampia gamma di chimiche degli analiti. Come dimostrato, gli amminoacidi non derivatizzati con catene laterali non polari, polari, con carica positiva e con carica negativa sono stati adeguatamente trattenuti e velocemente eluiti utilizzando un gradiente di 10 minuti. Questo gradiente ha inoltre separato i contaminanti del sistema degli amminoacidi dagli analiti target, aggiungendo robustezza al metodo. Seguendo questo metodo, i&nbsp;processi di preparazione del campione che richiedono molto tempo e una procedura elaborata \u2013 che possono anche includere costosi kit di derivatizzazione \u2013 vengono sostituiti da una semplice precipitazione delle proteine e dall\u2019analisi diretta dell\u2019estratto risultante. L\u2019analisi diretta degli amminoacidi non derivatizzati sulla colonna Raptor Polar X offre risultati eccellenti in un flusso di lavoro facile e veloce, rendendola un\u2019alternativa vantaggiosa rispetto ai metodi tradizionali.<\/p>\n\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><\/div><div class='chromatogram-article-inner-full'><div class=\"chromatogram-article-inner\">\n<div class=\"wp-block-custom-chromatogram-article\"><div class=\"wp-block-custom-chromatogram-article\"><div class=\"chromatogram-image regular-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_ff0579.png\" alt=\"Underivatized Amino Acids Analysis in Baby Formula on Raptor Polar X\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_FF0579<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks col-lg-6 col-12 peak-50\">\n<thead><tr><th><\/th><th style=\"text-align: left;width: 75px\">Peaks<\/th><th style=\"text-align: center;width: 75px\">t<sub>R<\/sub> (min)<\/th><th style=\"text-align: center;width: 75px\">Precursor Ion<\/th><th style=\"text-align: center;width: 75px\">Product Ion<\/th><\/tr><\/thead>\n<tbody><tr><td class=\"num\">1.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Tryptophan\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/54-12-6\/Tryptophan\" target=\"_blank\" rel=\"noopener\">Tryptophan<\/a><\/td><td class=\"oth\">1.17<\/td><td class=\"oth\">205.07<\/td><td class=\"oth\">146.08<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Phenylalanine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/63-91-2\/Phenylalanine\" target=\"_blank\" rel=\"noopener\">Phenylalanine<\/a><\/td><td class=\"oth\">1.26<\/td><td class=\"oth\">166.13<\/td><td class=\"oth\">120.10<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Leucine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/61-90-5\/Leucine\" target=\"_blank\" rel=\"noopener\">Leucine<\/a><\/td><td class=\"oth\">1.41<\/td><td class=\"oth\">132.13<\/td><td class=\"oth\">86.10<\/td><\/tr>\n<tr><td class=\"num\">4.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Isoleucine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/73-32-5\/Isoleucine\" target=\"_blank\" rel=\"noopener\">Isoleucine<\/a><\/td><td class=\"oth\">1.55<\/td><td class=\"oth\">132.13<\/td><td class=\"oth\">86.10<\/td><\/tr>\n<tr><td class=\"num\">5.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Methionine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/59-51-8\/Methionine\" target=\"_blank\" rel=\"noopener\">Methionine<\/a><\/td><td class=\"oth\">1.62<\/td><td class=\"oth\">150.07<\/td><td class=\"oth\">104.10<\/td><\/tr>\n<tr><td class=\"num\">6.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Tyrosine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/556-03-6\/Tyrosine\" target=\"_blank\" rel=\"noopener\">Tyrosine<\/a><\/td><td class=\"oth\">1.69<\/td><td class=\"oth\">182.10<\/td><td class=\"oth\">136.08<\/td><\/tr>\n<tr><td class=\"num\">7.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Taurine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/107-35-7\/Taurine\" target=\"_blank\" rel=\"noopener\">Taurine<\/a><\/td><td class=\"oth\">1.91<\/td><td class=\"oth\">126.07<\/td><td class=\"oth\">108.07<\/td><\/tr>\n<tr><td class=\"num\">8.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Valine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/516-06-3\/Valine\" target=\"_blank\" rel=\"noopener\">Valine<\/a><\/td><td class=\"oth\">1.98<\/td><td class=\"oth\">118.13<\/td><td class=\"oth\">72.11<\/td><\/tr>\n<tr><td class=\"num\">9.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Proline\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/609-36-9\/Proline\" target=\"_blank\" rel=\"noopener\">Proline<\/a><\/td><td class=\"oth\">2.29<\/td><td class=\"oth\">116.13<\/td><td class=\"oth\">70.09<\/td><\/tr>\n<tr><td class=\"num\">10.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Alanine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/302-72-7\/Alanine\" target=\"_blank\" rel=\"noopener\">Alanine<\/a><\/td><td class=\"oth\">3.00<\/td><td class=\"oth\">90.03<\/td><td class=\"oth\">44.10<\/td><\/tr>\n<\/tbody><\/table>\n<table class=\"peaks col-lg-6 col-12 peak-50\"><thead><tr><th><\/th><th style=\"text-align: left;width: 75px\">Peaks<\/th><th style=\"text-align: center;width: 75px\">t<sub>R<\/sub> (min)<\/th><th style=\"text-align: center;width: 75px\">Precursor Ion<\/th><th style=\"text-align: center;width: 75px\">Product Ion<\/th><\/tr><\/thead><tbody>\n<tr><td class=\"num\">11.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Threonine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/80-68-2\/Threonine\" target=\"_blank\" rel=\"noopener\">Threonine<\/a><\/td><td class=\"oth\">3.12<\/td><td class=\"oth\">120.13<\/td><td class=\"oth\">74.08<\/td><\/tr>\n<tr><td class=\"num\">12.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Glycine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/56-40-6\/Glycine\" target=\"_blank\" rel=\"noopener\">Glycine<\/a><\/td><td class=\"oth\">3.62<\/td><td class=\"oth\">76.10<\/td><td class=\"oth\">30.11<\/td><\/tr>\n<tr><td class=\"num\">13.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Glutamine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/56-85-9\/Glutamine\" target=\"_blank\" rel=\"noopener\">Glutamine<\/a><\/td><td class=\"oth\">3.87<\/td><td class=\"oth\">147.13<\/td><td class=\"oth\">84.07<\/td><\/tr>\n<tr><td class=\"num\">14.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Serine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/302-84-1\/Serine\" target=\"_blank\" rel=\"noopener\">Serine<\/a><\/td><td class=\"oth\">3.93<\/td><td class=\"oth\">106.07<\/td><td class=\"oth\">60.09<\/td><\/tr>\n<tr><td class=\"num\">15.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Asparagine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/70-47-3\/Asparagine\" target=\"_blank\" rel=\"noopener\">Asparagine<\/a><\/td><td class=\"oth\">4.08<\/td><td class=\"oth\">133.13<\/td><td class=\"oth\">74.07<\/td><\/tr>\n<tr><td class=\"num\">16.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Arginine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/74-79-3\/Arginine\" target=\"_blank\" rel=\"noopener\">Arginine<\/a><\/td><td class=\"oth\">4.47<\/td><td class=\"oth\">175.17<\/td><td class=\"oth\">70.09<\/td><\/tr>\n<tr><td class=\"num\">17.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Histidine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/71-00-1\/Histidine\" target=\"_blank\" rel=\"noopener\">Histidine<\/a><\/td><td class=\"oth\">4.66<\/td><td class=\"oth\">156.07<\/td><td class=\"oth\">110.16<\/td><\/tr>\n<tr><td class=\"num\">18.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Lysine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/56-87-1\/Lysine\" target=\"_blank\" rel=\"noopener\">Lysine<\/a><\/td><td class=\"oth\">4.97<\/td><td class=\"oth\">147.13<\/td><td class=\"oth\">84.13<\/td><\/tr>\n<tr><td class=\"num\">19.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Glutamic acid\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/617-65-2\/Glutamic acid\" target=\"_blank\" rel=\"noopener\">Glutamic acid<\/a><\/td><td class=\"oth\">5.89<\/td><td class=\"oth\">148.10<\/td><td class=\"oth\">84.10<\/td><\/tr>\n<tr><td class=\"num\">20.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Cystine\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/56-89-3\/Cystine\" target=\"_blank\" rel=\"noopener\">Cystine<\/a><\/td><td class=\"oth\">6.10<\/td><td class=\"oth\">241.13<\/td><td class=\"oth\">152.00<\/td><\/tr>\n<tr><td class=\"num\">21.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Aspartic acid\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/617-45-8\/Aspartic acid\" target=\"_blank\" rel=\"noopener\">Aspartic acid<\/a><\/td><td class=\"oth\">7.12<\/td><td class=\"oth\">134.07<\/td><td class=\"oth\">74.06<\/td><\/tr>\n<\/tbody><\/table><\/div><div class=\"chromatogram-conditions\"><h4>Conditions<\/h4><div class=\"conditions-container container-fluid\"><div class=\"row\"><table class=\"conditions col-lg-6 col-12\"><tr><th class=\"conditions_header\" scope=\"row\">Column<\/th><td>Raptor Polar X  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9311A12?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_FF0579\" rel=\"noopener\">cat.# 9311A12<\/a>)<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Dimensions:<\/th><td>100 mm x 2.1 mm ID<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Particle Size:<\/th><td>2.7 \u00b5m<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Temp.:<\/th><td>30 \u00b0C<\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Standard\/Sample<\/th><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Diluent:<\/th><td>20:80 Water:acetonitrile, 0.01 N HCl<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Conc.:<\/th><td>  Endogenous amino acids<\/td><\/tr><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Inj. Vol.:<\/th><td>5 \u00b5L <\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Mobile Phase<\/th><td><\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">A:<\/th><td>Water, 0.5% formic acid <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>Mobile Phase B: 9:1 Acetonitrile:20 mM ammonium formate in water (pH 3.0) (The ammonium formate concentration is 20 mM relative to the total volume of mobile phase B. See preparation notes for instructions on diluting a 200 mM aqueous starting solution.) <\/td><\/tr><tr><td><\/td><td><table class=\"cgram_ramp\"><thead><tr><th>Time (min)<\/th><th>Flow (mL\/min)<\/th><th>%A<\/th><th>%B<\/th><\/tr><\/thead><tbody><tr><td>0.00<\/td><td>0.5<\/td><td>12<\/td><td>88<\/td><\/tr><tr><td>3.50<\/td><td>0.5<\/td><td>12<\/td><td>88<\/td><\/tr><tr><td>8.00<\/td><td>0.5<\/td><td>70<\/td><td>30<\/td><\/tr><tr><td>8.01<\/td><td>0.5<\/td><td>12<\/td><td>88<\/td><\/tr><tr><td>10.0<\/td><td>0.5<\/td><td>12<\/td><td>88<\/td><\/tr><\/tbody><\/table><\/td><\/tr><\/table><table class=\"conditions col-lg-6 col-12\"><tr><th class=\"conditions_header\" scope=\"row\">Detector<\/th><td>MS\/MS<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Ion Mode:<\/th><td>ESI+ <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Mode:<\/th><td>MRM <\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Instrument<\/th><td>UHPLC<\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Sample Preparation<\/th><td>A 200 \u00b5L aliquot of protein hydrolysate formula (Similac ALIMENTUM) was mixed with 800 \u00b5L of acetonitrile and 10 \u00b5L of 1 N HCl. After centrifugation at 4000 rpm for 5 minutes, the supernatant was diluted 20-fold with 20:80 water:acetonitrile (0.01 N HCl) and injected for analysis.<\/td><\/tr><tr class=\"cgram_header_row\"><th class=\"conditions_header\" scope=\"row\">Notes<\/th><td><i>Mobile Phase B Preparation:<\/i> To make 500 mL of mobile phase B, measure ~45 mL of water into a small beaker and add 1 mL of 10 M ammonium formate solution. Adjust pH to 3.0 by adding formic acid and then bring the volume to 50 mL with water. Combine this 50 mL ammonium formate solution (pH 3.0) with 450 mL of acetonitrile to complete the preparation.   <\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_ff0579.pdf\" target=\"_blank\" rel=\"noopener noreferrer\"><svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"18\" height=\"18\" viewBox=\"0 0 18 18\"><g data-name=\"Group 2996\"><path data-name=\"Rectangle 1246\" d=\"M0 0h18v18H0z\" style=\"fill: none;\"><\/path><\/g><g data-name=\"Group 2997\"><path data-name=\"Path 729\" d=\"M13.412 11.4v2.017H5.345V11.4H4v2.017a1.349 1.349 0 0 0 1.345 1.345h8.068a1.349 1.349 0 0 0 1.345-1.345V11.4zm-.672-2.694-.948-.948-1.741 1.735V4H8.706v5.493L6.965 7.758l-.948.948 3.361 3.361z\" transform=\"translate(-.437 -.414)\" style=\"fill: rgb(13, 123, 196);\"><\/path><\/g><\/svg>Download PDF<\/a><\/div><\/div><\/div>\n<\/div><\/div><\/div>\n\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Questa analisi diretta LC-MS\/MS degli amminoacidi non derivatizzati nelle formule per lattanti \u00e8 un semplice flusso di lavoro workflow che fornisce una buona ritenzione e una rapida eluizione di una vasta gamma di aminoacidi.<\/p>\n","protected":false},"author":11,"featured_media":90022,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_kadence_starter_templates_imported_post":false,"_kad_post_transparent":"","_kad_post_title":"","_kad_post_layout":"","_kad_post_sidebar_id":"","_kad_post_content_style":"","_kad_post_vertical_padding":"","_kad_post_feature":"","_kad_post_feature_position":"","_kad_post_header":false,"_kad_post_footer":false,"footnotes":""},"categories":[788],"tags":[],"industries-application":[2290,2259],"post-badge":[],"resource-type":[2768],"product-library":[2535,2517],"resource-technique":[2334,2364],"ppma_author":[414],"class_list":["post-43562","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-articoli","industries-application-food-beverage-it","industries-application-sicurezza-alimentare","resource-type-featured-application","product-library-colonne-lc","product-library-prodotti-per-cromatografia-liquida","resource-technique-cromatografia-liquida-lc","resource-technique-ms-ms-it"],"acf":[],"taxonomy_info":{"category":[{"value":788,"label":"Articoli"}],"industries-application":[{"value":2290,"label":"Food &amp; 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