{"id":43342,"date":"2020-10-08T00:00:00","date_gmt":"2020-10-08T00:00:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/a-fast-dilute-and-shoot-method-for-simultaneous-5-hydroxyindoleacetic-acid-5-hiaa-vanillylmandelic-acid-vma-and-homovanillic-acid-hva-lc-msms-analysis-in-human-urine\/"},"modified":"2025-12-10T15:14:11","modified_gmt":"2025-12-10T15:14:11","slug":"a-fast-dilute-and-shoot-method-for-simultaneous-5-hydroxyindoleacetic-acid-5-hiaa-vanillylmandelic-acid-vma-and-homovanillic-acid-hva-lc-msms-analysis-in-human-urine","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/fr\/application-notes-it\/cfan2465\/a-fast-dilute-and-shoot-method-for-simultaneous-5-hydroxyindoleacetic-acid-5-hiaa-vanillylmandelic-acid-vma-and-homovanillic-acid-hva-lc-msms-analysis-in-human-urine","title":{"rendered":"A Fast Dilute-And-Shoot Method for Simultaneous 5-Hydroxyindoleacetic Acid (5-HIAA), Vanillylmandelic Acid (VMA), and Homovanillic Acid (HVA) LC-MS\/MS Analysis in Human Urine"},"content":{"rendered":"\n<h2 class=\"wp-block-heading\">Abstract<\/h2>\n\n\n\n<p>In this method evaluation study, we demonstrated the performance of a fast dilute-and-shoot LC-MS\/MS method using a Raptor Biphenyl column for the simultaneous analysis of 5-hydroxyindoleacetic acid (5-HIAA), vanillylmandelic acid (VMA), and homovanillic acid (HVA) in human urine. Excellent results were obtained for linearity, accuracy, and precision. With this method, these serotonin and catecholamine metabolites can be measured in human urine with a 5-minute total analysis time. The analytical method is applicable to clinical labs that require a combination of fast analysis times and low-cost sample preparation procedures for 5-HIAA, VMA, and HVA using LC-MS\/MS analysis.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Introduction<\/h2>\n\n\n\n<p>5-Hydroxyindoleacetic acid (5-HIAA), vanillylmandelic acid (VMA), and homovanillic acid (HVA) are the final metabolites of serotonin, epinephrine, and dopamine, respectively (Figure 1). Measurement of these metabolites in urine is used for the diagnosis of carcinoid and neuroendocrine tumors including neuroblastoma, pheochromocytoma, and paraganglioma. Several commercial products are available for the simultaneous measurement of these three analytes in urine. However, these commercial kits all require extensive sample preparation and chromatographic analysis times of up to 16 minutes. The development of a faster and simpler method would greatly reduce the time and cost of this type of clinical test. In this study, we developed a fast dilute-and-shoot method for accurate and specific 5-HIAA, VMA, and HVA LC-MS\/MS analysis in urine using a Raptor Biphenyl column. The Raptor Biphenyl column was selected for this work because our initial investigation determined that it offers better peak shape and sensitivity than a C18 column.<\/p>\n\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><strong>Figure 1:<\/strong>\u00a0Structures of VMA, HVA, and 5-HIAA.<\/div><div class='chromatogram-article-inner-full'><div class=\"chromatogram-article-inner\">\n<style>.kb-image43342_0a9a6e-09 .kb-image-has-overlay:after{opacity:0.3;}<\/style>\n<div class=\"wp-block-kadence-image kb-image43342_0a9a6e-09\"><figure class=\"aligncenter size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"453\" src=\"https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-01-1024x453.jpg\" alt=\"\" class=\"kb-img wp-image-18868\" title=\"-\" srcset=\"https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-01-1024x453.jpg 1024w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-01-300x133.jpg 300w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-01-768x340.jpg 768w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-01-1536x679.jpg 1536w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-01.jpg 1800w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/figure><\/div>\n\n<\/div><\/div><\/div>\n\n\n<div style=\"height:20px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Experimental<\/h2>\n\n\n\n<p>To evaluate linearity, eight standard solutions (0.2, 0.4, 1.0, 5.0, 10, 25, 50, and 100 \u00b5g\/mL) were prepared in synthetic human urine (Surine negative urine control, Cerilliant). In addition, two levels of urine control samples (Bio-Rad Lyphochek quantitative urine control, bilevel minipak #375X) were used as QC samples for accuracy and precision analysis. Controls 1 and 2 were described by Bio-Rad Laboratories as normal and abnormal levels, respectively, and had labeled concentration ranges (Table II) for 5-HIAA, VMA, and HVA that were determined by Bio-Rad using a reversed-phase HPLC method.<\/p>\n\n\n\n<p>All prepared standards and samples were diluted 10x by mixing 40 \u00b5L of prepared standard or sample in urine with 360 \u00b5L of water and 10 \u00b5L of internal standard solution (33.3 \u00b5g\/mL of 5-HIAA-D5, VMA-D3, and HVA-D5 in methanol) in a 0.45 \u00b5m Thomson SINGLE StEP PVDF filter vial (Restek, cat.# 25896). Standards and samples were filtered and injected for analysis.<\/p>\n\n\n\n<p>5-HIAA, VMA, and HVA LC analysis was performed on a Shimadzu Nexera XR System coupled with a Sciex API 4000 mass spectrometer. Instrument conditions were as follows and analyte transitions are provided in Table I.<\/p>\n\n\n\n<figure class=\"wp-block-table\"><table style=\"border-style:none;border-width:0px\"><tbody><tr><td>Analytical column:<\/td><td colspan=\"2\">Raptor Biphenyl (5 \u00b5m, 100 mm x 2.1 mm;&nbsp;cat.# 9309512)<\/td><\/tr><tr><td>Guard column:<\/td><td colspan=\"2\">Raptor Biphenyl EXP guard column cartridge (5 \u00b5m, 5 mm x 2.1 mm;<br>cat.# 930950252) and EXP direct connect holder (cat.# 25808)<\/td><\/tr><tr><td>Mobile phase A:<\/td><td colspan=\"2\">0.1% Formic acid + 5 mM ammonium formate in water<\/td><\/tr><tr><td>Mobile phase B:<\/td><td colspan=\"2\">Methanol<\/td><\/tr><tr><td>Gradient<\/td><td>Time (min)<\/td><td>%B<\/td><\/tr><tr><td>&nbsp;<\/td><td>0.00<\/td><td>15<\/td><\/tr><tr><td>&nbsp;<\/td><td>3.00<\/td><td>80<\/td><\/tr><tr><td>&nbsp;<\/td><td>3.01<\/td><td>15<\/td><\/tr><tr><td>&nbsp;<\/td><td>5.00<\/td><td>15<\/td><\/tr><tr><td>Flow rate:<\/td><td>0.5 mL\/min<\/td><td>&nbsp;<\/td><\/tr><tr><td>Injection volume:<\/td><td>5 \u00b5L<\/td><td>&nbsp;<\/td><\/tr><tr><td>Column temp.:&nbsp;<\/td><td>30 \u00b0C<\/td><td>&nbsp;<\/td><\/tr><tr><td>Ion mode:<\/td><td>Negative ESI<\/td><td>&nbsp;<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<div style=\"height:20px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<p><strong>Table I<\/strong>: Analyte Transitions<\/p>\n\n\n\n<figure class=\"wp-block-table\"><table class=\"has-fixed-layout\"><tbody><tr><td><strong>Analyte<\/strong><\/td><td><strong>CAS#<\/strong><\/td><td><strong>Precursor Ion<\/strong><\/td><td><strong>Product Ion Quantifier<\/strong><\/td><td><strong>Product Ion Qualifier<\/strong><\/td><\/tr><tr><td>5-HIAA-D5<\/td><td>\u2014<\/td><td>195.1<\/td><td>148.0<\/td><td>\u2014<\/td><\/tr><tr><td>5-HIAA<\/td><td>54-16-0<\/td><td>190.0<\/td><td>145.9<\/td><td>144.0<\/td><\/tr><tr><td>VMA-D3<\/td><td>\u2014<\/td><td>200.1<\/td><td>139.9<\/td><td>\u2014<\/td><\/tr><tr><td>VMA<\/td><td>55-10-7<\/td><td>197.0<\/td><td>137.9<\/td><td>136.9<\/td><\/tr><tr><td>HVA-D5<\/td><td>\u2014<\/td><td>186.1<\/td><td>127.0<\/td><td>\u2014<\/td><\/tr><tr><td>HVA<\/td><td>306-08-1<\/td><td>181.0<\/td><td>121.9<\/td><td>136.9<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<h2 class=\"wp-block-heading\">Results and Discussion<\/h2>\n\n\n\n<p>To develop a quantitative dilute-and-shoot method, it is important to ensure chromatographic separation of all matrix interferences from the target analytes. Upon initial method development, it was observed that the response of HVA internal standard suffered from urine matrix suppression and resulted in inaccurate measurement of HVA concentration. To solve this problem, it was necessary to modify the additive type in the mobile phase and select an LC column with the proper phase chemistry. The final method demonstrated that the Raptor Biphenyl column is unique in that it offered excellent chromatographic performance for simultaneous accurate and precise quantification of 5-HIAA, VMA, and HVA in diluted human urine samples (see Figure 3, 4, and 5 for representative chromatograms).<\/p>\n\n\n\n<p>Linear responses were obtained from 0.2\u2013100 \u00b5g\/mL for 5-HIAA, VMA, and HVA. A 1\/x weighted quadratic regression provided the best fit for the standard calibration curves of all three analytes. The deviations were all \u226410% (except for the lowest concentration, which was \u226415%) and the R<sup>2<\/sup>&nbsp;values all were 0.999\u20131.000 (see examples in Figure 2). As estimated from the signal-to-noise value of the 0.2 \u00b5g\/mL standard injection, the LODs were 0.03, 0.007, and 0.02 \u00b5g\/mL for 5-HIAA, VMA, and HVA, respectively.<\/p>\n\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><strong>Figure 2:<\/strong>\u00a0Calibration Curves for 5-HIAA, VMA, and HVA LC Analyses.<\/div><div class='chromatogram-article-inner-full'><div class=\"chromatogram-article-inner\">\n<div class=\"wp-block-image\">\n<figure class=\"aligncenter size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"570\" height=\"1024\" src=\"https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-570x1024.jpg\" alt=\"\" class=\"wp-image-18874\" title=\"-\" srcset=\"https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-570x1024.jpg 570w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-167x300.jpg 167w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-768x1381.jpg 768w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-854x1536.jpg 854w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-1139x2048.jpg 1139w, https:\/\/discover.restek.com\/wp-content\/uploads\/figure-article-CFAN2465-02-scaled.jpg 1424w\" sizes=\"auto, (max-width: 570px) 100vw, 570px\" \/><\/figure>\n<\/div>\n<\/div><\/div><\/div>\n\n\n<p>Three sets of samples were prepared and analyzed to evaluate method accuracy and precision. Method accuracy was demonstrated to be acceptable as the quantitative concentrations fell within the nominal concentration ranges for both the low and high urine control samples (Table II). The %RSD ranged from 0.2\u20132.8% and 0.2\u20132.9% for intraday and interday, respectively (data not shown), indicating acceptable method precision.<\/p>\n\n\n\n<p><strong>Table II<\/strong>: Accuracy and Precision of Urine Control Samples<\/p>\n\n\n\n<figure class=\"wp-block-table\"><table class=\"has-fixed-layout\"><tbody><tr><td>&nbsp;<\/td><td colspan=\"3\"><strong>QC Level 1 (n = 3)<\/strong><\/td><td colspan=\"3\"><strong>QC Level 2 (n = 3)<\/strong><\/td><\/tr><tr><td><strong>Analyte<\/strong><\/td><td><strong>Range (\u00b5g\/mL)*<\/strong><\/td><td><strong>Average Conc. (\u00b5g\/mL)<\/strong><\/td><td><strong>%RSD<\/strong><\/td><td><strong>Range (\u00b5g\/mL)*<\/strong><\/td><td><strong>Average Conc. (\u00b5g\/mL)<\/strong><\/td><td><strong>%RSD<\/strong><\/td><\/tr><tr><td>5-HIAA<\/td><td>2.69\u20134.49<\/td><td>3.59<\/td><td>2.2<\/td><td>21.6\u201332.7<\/td><td>27.5<\/td><td>1.7<\/td><\/tr><tr><td>VMA<\/td><td>2.12\u20133.90<\/td><td>3.34<\/td><td>1.0<\/td><td>10.9\u201318.2<\/td><td>16.3<\/td><td>0.2<\/td><\/tr><tr><td>HVA<\/td><td>1.33\u20132.46<\/td><td>1.95<\/td><td>2.4<\/td><td>12.9\u201318.9<\/td><td>17.5<\/td><td>2.9<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<p>*The QC sample concentration ranges were supplied by the vendor.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">Conclusion<\/h2>\n\n\n\n<p>Commercial kits for the analysis of 5-hydroxyindoleacetic acid (5-HIAA), vanillylmandelic acid (VMA), and homovanillic acid (HVA), require extensive sample preparation and chromatographic analysis times up to 16 minutes. In contrast, using Restek\u2019s dilute-and-shoot method and a Raptor Biphenyl column, these serotonin and catecholamine metabolites can be measured in a quick 5-minute analysis. This method is applicable to clinical 5-HIAA, VMA, and HVA LC-MS\/MS analysis in urine and provides the benefits of both fast analysis times and low-cost sample preparation procedures.<\/p>\n\n\n<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><strong>Figure 3:<\/strong>\u00a05-HIAA, VMA, and HVA LC Analysis in Fortified Synthetic Human Urine.<\/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 wide-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0660.png\" alt=\"Vanillylmandelic Acid, Homovanillic Acid, and 5-Hydroxyindoleacetic Acid in Synthetic Human Urine on Raptor Biphenyl\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_CF0660<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks col-lg-6 col-12\">\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\">Conc.<br \/>(\u00b5g\/mL)<\/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\">4-Hydroxy-3-methoxymandelic acid-D3 (VMA-D3)<\/td><td class=\"oth\">0.90<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">200.1<\/td><td class=\"oth\">139.9<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Vanillylmandelic acid (VMA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/55-10-7\/Vanillylmandelic acid\" target=\"_blank\" rel=\"noopener\">Vanillylmandelic acid (VMA)<\/a><\/td><td class=\"oth\">0.92<\/td><td class=\"oth\">1<\/td><td class=\"oth\">197.0<\/td><td class=\"oth\">137.9<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\">5-Hydroxyindole-4,6,7-D3-3-acetic-D2 acid (5-HIAA-D5)<\/td><td class=\"oth\">1.80<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">195.1<\/td><td class=\"oth\">148.0<\/td><\/tr>\n<tr><td class=\"num\">4.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for 5-Hydroxyindole-3-acetic acid (5-HIAA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/54-16-0\/5-Hydroxyindole-3-acetic acid\" target=\"_blank\" rel=\"noopener\">5-Hydroxyindole-3-acetic acid (5-HIAA)<\/a><\/td><td class=\"oth\">1.82<\/td><td class=\"oth\">1<\/td><td class=\"oth\">190.0<\/td><td class=\"oth\">145.9<\/td><\/tr>\n<tr><td class=\"num\">5.<\/td><td class=\"cmpd\">4-Hydroxy-3-methoxyphenyl-D3-acetic-D2 acid (HVA-D5)<\/td><td class=\"oth\">2.10<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">186.1<\/td><td class=\"oth\">127.0<\/td><\/tr>\n<tr><td class=\"num\">6.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Homovanillic acid (HVA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/306-08-1\/Homovanillic acid\" target=\"_blank\" rel=\"noopener\">Homovanillic acid (HVA)<\/a><\/td><td class=\"oth\">2.12<\/td><td class=\"oth\">1<\/td><td class=\"oth\">181.0<\/td><td class=\"oth\">121.9<\/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 Biphenyl  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9309512?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0660\" rel=\"noopener\">cat.# 9309512<\/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>5 \u00b5m<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Pore Size:<\/th><td>90 \u00c5<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Guard Column:<\/th><td>Raptor Biphenyl guard column cartridge 5 mm, 2.1 mm ID, 5 \u00b5m (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/930950252?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0660\" rel=\"noopener\">cat.# 930950252<\/a>)<\/td><\/tr><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>Water<\/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>0.1% Formic acid, 5 mM ammonium formate in water <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>Methanol <\/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>85<\/td><td>15<\/td><\/tr><tr><td>3.00<\/td><td>0.5<\/td><td>20<\/td><td>80<\/td><\/tr><tr><td>3.01<\/td><td>0.5<\/td><td>85<\/td><td>15<\/td><\/tr><tr><td>5.00<\/td><td>0.5<\/td><td>85<\/td><td>15<\/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>The synthetic urine (Surine) was fortified with VMA, HVA, and 5-HIAA to a concentration of 10 \u03bcg\/mL. A 40 \u03bcL aliquot of fortified urine was mixed with 360 \u03bcL of water and 10 \u03bcL of internal standard solution (33.3 \u03bcg\/mL in methanol) in a Thomson 0.45 \u03bcm PVDF filter vial and injected for analysis after filtration.<\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0660.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<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><strong>Figure 4:<\/strong>\u00a05-HIAA, VMA, and HVA LC Analysis in Human Urine (Control Level 1).<\/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 wide-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0661.png\" alt=\"Vanillylmandelic Acid, Homovanillic Acid, and 5-Hydroxyindoleacetic Acid in Human Urine (Quantitative Control Level 1) on Raptor Biphenyl\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_CF0661<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks col-lg-6 col-12\">\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\">Conc.<br \/>(\u00b5g\/mL)<\/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\">4-Hydroxy-3-methoxymandelic acid-D3 (VMA-D3)<\/td><td class=\"oth\">0.90<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">200.1<\/td><td class=\"oth\">139.9<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Vanillylmandelic acid (VMA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/55-10-7\/Vanillylmandelic acid\" target=\"_blank\" rel=\"noopener\">Vanillylmandelic acid (VMA)<\/a><\/td><td class=\"oth\">0.92<\/td><td class=\"oth\">2.12\u20133.90*<\/td><td class=\"oth\">197.0<\/td><td class=\"oth\">137.9<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\">5-Hydroxyindole-4,6,7-D3-3-acetic-D2 acid (5-HIAA-D5)<\/td><td class=\"oth\">1.80<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">195.1<\/td><td class=\"oth\">148.0<\/td><\/tr>\n<tr><td class=\"num\">4.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for 5-Hydroxyindole-3-acetic acid (5-HIAA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/54-16-0\/5-Hydroxyindole-3-acetic acid\" target=\"_blank\" rel=\"noopener\">5-Hydroxyindole-3-acetic acid (5-HIAA)<\/a><\/td><td class=\"oth\">1.82<\/td><td class=\"oth\">2.69\u20134.49*<\/td><td class=\"oth\">190.0<\/td><td class=\"oth\">145.9<\/td><\/tr>\n<tr><td class=\"num\">5.<\/td><td class=\"cmpd\">4-Hydroxy-3-methoxyphenyl-D3-acetic-D2 acid (HVA-D5)<\/td><td class=\"oth\">2.10<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">186.1<\/td><td class=\"oth\">127.0<\/td><\/tr>\n<tr><td class=\"num\">6.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Homovanillic acid (HVA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/306-08-1\/Homovanillic acid\" target=\"_blank\" rel=\"noopener\">Homovanillic acid (HVA)<\/a><\/td><td class=\"oth\">2.12<\/td><td class=\"oth\">1.33\u20132.46*<\/td><td class=\"oth\">181.0<\/td><td class=\"oth\">121.9<\/td><\/tr>\n<\/tbody><\/table><div class=\"peaknotes\">*These concentration ranges were supplied by the vendor (see notes).<\/div><\/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 Biphenyl  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9309512?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0661\" rel=\"noopener\">cat.# 9309512<\/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>5 \u00b5m<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Pore Size:<\/th><td>90 \u00c5<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Guard Column:<\/th><td>Raptor Biphenyl guard column cartridge 5 mm, 2.1 mm ID, 5 \u00b5m (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/930950252?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0661\" rel=\"noopener\">cat.# 930950252<\/a>)<\/td><\/tr><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>Water<\/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>0.1% Formic acid, 5 mM ammonium formate in water <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>Methanol <\/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>85<\/td><td>15<\/td><\/tr><tr><td>3.00<\/td><td>0.5<\/td><td>20<\/td><td>80<\/td><\/tr><tr><td>3.01<\/td><td>0.5<\/td><td>85<\/td><td>15<\/td><\/tr><tr><td>5.00<\/td><td>0.5<\/td><td>85<\/td><td>15<\/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>The human urine sample is the Bio-Rad Lyphochek quantitative urine control, level 1. A 40 \u03bcL aliquot of urine was mixed with 360 \u03bcL of water and 10 \u03bcL of internal standard solution (33.3 \u03bcg\/mL in methanol) in a Thomson 0.45 \u03bcm PVDF filter vial and injected for analysis after filtration. <\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0661.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<div class=\"wp-block-custom-chromatogram-article-top\"><div class=\"chromatogram-article-placeholder\"><div class=\"figure-heading\"><strong>Figure 5:<\/strong>\u00a05-HIAA, VMA, and HVA LC Analysis in Human Urine (Control Level 2)<\/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 wide-image\"><img decoding=\"async\" src=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0662.png\" alt=\"Vanillylmandelic Acid, Homovanillic Acid, and 5-Hydroxyindoleacetic Acid in Human Urine (Quantitative Control Level 2) on Raptor Biphenyl\" title=\"-\"><\/div><p class=\"article-id\" style=\"text-align:center\"> LC_CF0662<\/p><div class=\"chromatogram-peaks\"><h4>Peaks<\/h4><table class=\"peaks col-lg-6 col-12\">\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\">Conc.<br \/>(\u00b5g\/mL)<\/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\">4-Hydroxy-3-methoxymandelic acid-D3 (VMA-D3)<\/td><td class=\"oth\">0.90<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">200.1<\/td><td class=\"oth\">139.9<\/td><\/tr>\n<tr><td class=\"num\">2.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Vanillylmandelic acid (VMA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/55-10-7\/Vanillylmandelic acid\" target=\"_blank\" rel=\"noopener\">Vanillylmandelic acid (VMA)<\/a><\/td><td class=\"oth\">0.91<\/td><td class=\"oth\">10.9\u201318.2*<\/td><td class=\"oth\">197.0<\/td><td class=\"oth\">137.9<\/td><\/tr>\n<tr><td class=\"num\">3.<\/td><td class=\"cmpd\">5-Hydroxyindole-4,6,7-D3-3-acetic-D2 acid (5-HIAA-D5)<\/td><td class=\"oth\">1.81<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">195.1<\/td><td class=\"oth\">148.0<\/td><\/tr>\n<tr><td class=\"num\">4.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for 5-Hydroxyindole-3-acetic acid (5-HIAA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/54-16-0\/5-Hydroxyindole-3-acetic acid\" target=\"_blank\" rel=\"noopener\">5-Hydroxyindole-3-acetic acid (5-HIAA)<\/a><\/td><td class=\"oth\">1.83<\/td><td class=\"oth\">21.6\u201332.7*<\/td><td class=\"oth\">190.0<\/td><td class=\"oth\">145.9<\/td><\/tr>\n<tr><td class=\"num\">5.<\/td><td class=\"cmpd\">4-Hydroxy-3-methoxyphenyl-D3-acetic-D2 acid (HVA-D5)<\/td><td class=\"oth\">2.11<\/td><td class=\"oth\">0.83<\/td><td class=\"oth\">186.1<\/td><td class=\"oth\">127.0<\/td><\/tr>\n<tr><td class=\"num\">6.<\/td><td class=\"cmpd\"><a class=\"cmpd_link\" title=\"View compound information for Homovanillic acid (HVA)\" href=\"https:\/\/ez.restek.com\/compound\/view\/en\/306-08-1\/Homovanillic acid\" target=\"_blank\" rel=\"noopener\">Homovanillic acid (HVA)<\/a><\/td><td class=\"oth\">2.14<\/td><td class=\"oth\">12.9\u201318.9*<\/td><td class=\"oth\">181.0<\/td><td class=\"oth\">121.9<\/td><\/tr>\n<\/tbody><\/table><div class=\"peaknotes\">*These concentration ranges were supplied by the vendor (see notes).<\/div><\/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 Biphenyl  (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/9309512?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0662\" rel=\"noopener\">cat.# 9309512<\/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>5 \u00b5m<\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">Pore Size:<\/th><td>90 \u00c5<\/td><\/tr><tr><td><\/td><tr><th class=\"sub conditions_header\" scope=\"row\">Guard Column:<\/th><td>Raptor Biphenyl guard column cartridge 5 mm, 2.1 mm ID, 5 \u00b5m (<a target=\"_blank\" href=\"https:\/\/www.restek.com\/p\/930950252?utm_source=chromatograms&amp;utm_medium=link&amp;utm_campaign=LC_CF0662\" rel=\"noopener\">cat.# 930950252<\/a>)<\/td><\/tr><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>Water<\/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>0.1% Formic acid, 5 mM ammonium formate in water <\/td><\/tr><tr><th class=\"sub conditions_header\" scope=\"row\">B:<\/th><td>Methanol <\/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>85<\/td><td>15<\/td><\/tr><tr><td>3.00<\/td><td>0.5<\/td><td>20<\/td><td>80<\/td><\/tr><tr><td>3.01<\/td><td>0.5<\/td><td>85<\/td><td>15<\/td><\/tr><tr><td>5.00<\/td><td>0.5<\/td><td>85<\/td><td>15<\/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>The human urine sample is the Bio-Rad Lyphochek quantitative urine control, level 2. A 40 \u03bcL aliquot of urine was mixed with 360 \u03bcL of water and 10 \u03bcL of internal standard solution (33.3 \u03bcg\/mL in methanol) in a Thomson 0.45 \u03bcm PVDF filter vial and injected for analysis after filtration. <\/td><\/tr><\/table><\/div><\/div><\/div><div class=\"chromatogram-pdf-link\"><a href=\"https:\/\/ez.restek.com\/images\/cgram\/lc_cf0662.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:20px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n        <div class=\"cpb\">\n            <h3 class=\"cpb-heading\">Products Mentioned<\/h3>\n            <hr class=\"cpb-heading-underline\" \/>\n            <div class=\"cpb-list\">\n                                    <div class=\"cpb-item\">\n                        <div class=\"cpb-col cpb-col--left\">\n                            <a class=\"cpb-catalog\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/9309512\">                                Catalog No. 9309512                            <\/a>                        <\/div>\n                        <div class=\"cpb-col cpb-col--middle\">\n                            <div class=\"cpb-title\">Colonnes Raptor Biphenyl, 5\u00b5m 100 x 2,1mm<\/div>\n                        <\/div>\n                        <div class=\"cpb-col cpb-col--right\">\n                                                            <a class=\"cpb-view-btn\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/9309512\">Voir le produit<\/a>\n                                                    <\/div>\n                    <\/div>\n                                    <div class=\"cpb-item\">\n                        <div class=\"cpb-col cpb-col--left\">\n                            <a class=\"cpb-catalog\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/930950252\">                                Catalog No. 930950252                            <\/a>                        <\/div>\n                        <div class=\"cpb-col cpb-col--middle\">\n                            <div class=\"cpb-title\">Cartouche de garde Raptor Biphenyl EXP, 5 \u00b5m, L 5\u00a0x DI 2.1 mm, lot de 3<\/div>\n                        <\/div>\n                        <div class=\"cpb-col cpb-col--right\">\n                                                            <a class=\"cpb-view-btn\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/930950252\">Voir le produit<\/a>\n                                                    <\/div>\n                    <\/div>\n                                    <div class=\"cpb-item\">\n                        <div class=\"cpb-col cpb-col--left\">\n                            <a class=\"cpb-catalog\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/25808\">                                Catalog No. 25808                            <\/a>                        <\/div>\n                        <div class=\"cpb-col cpb-col--middle\">\n                            <div class=\"cpb-title\">Support de cartouche \u00e0 connexion directe EXP, livr\u00e9 avec un \u00e9crou et 2 ferrules<\/div>\n                        <\/div>\n                        <div class=\"cpb-col cpb-col--right\">\n                                                            <a class=\"cpb-view-btn\" target=\"_blank\" rel=\"noopener noreferrer\" href=\"https:\/\/www.restek.com\/p\/25808\">Voir le produit<\/a>\n                                                    <\/div>\n                    <\/div>\n                            <\/div>\n        <\/div>\n        \n\n\n<div style=\"height:100px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Using Restek\u2019s dilute-and-shoot method and a Raptor Biphenyl column, serotonin and catecholamine metabolites can be measured simultaneously in a quick 5-minute analysis. This method is applicable to clinical 5-HIAA, VMA, and HVA LC analysis in urine and provides the benefit of both fast analysis times and low-cost sample preparation procedures.<\/p>\n","protected":false},"author":46,"featured_media":6325,"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":[784],"tags":[],"industries-application":[2245,2246,2247],"post-badge":[],"resource-type":[],"product-library":[2535,2517,2519],"resource-technique":[2334,2364],"ppma_author":[578,752],"class_list":["post-43342","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-application-notes-it","industries-application-clinico","industries-application-diagnostica-clinica","industries-application-ricerca-clinica","product-library-colonne-lc","product-library-prodotti-per-cromatografia-liquida","product-library-prodotti-per-la-preparazione-del-campione","resource-technique-cromatografia-liquida-lc","resource-technique-ms-ms-it"],"acf":[],"taxonomy_info":{"category":[{"value":784,"label":"Application 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