{"id":1193,"date":"2024-04-11T14:43:54","date_gmt":"2024-04-11T11:43:54","guid":{"rendered":"https:\/\/sisu.ut.ee\/lcms_method_validation\/self-test-10-2\/"},"modified":"2024-04-11T14:43:54","modified_gmt":"2024-04-11T11:43:54","slug":"self-test-10-2","status":"publish","type":"page","link":"https:\/\/sisu.ut.ee\/lcms_method_validation\/self-test-10-2\/","title":{"rendered":"Self test 10.2"},"content":{"rendered":"<div id=\"watupro_quiz\" class=\"quiz-area single-page-quiz\">\n<p id=\"submittingExam26\" style=\"display:none;text-align:center;\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/sisu.ut.ee\/wp-content\/plugins\/watupro\/img\/loading.gif\" width=\"16\" height=\"16\"><\/p>\n\n\n\n<form action=\"\" method=\"post\" class=\"quiz-form\" id=\"quiz-26\" enctype=\"multipart\/form-data\">\n<div class=\"watu-question \" id=\"question-1\" style=\";\"><div id=\"questionWrap-1\" class=\"   watupro-question-id-107\">\n\t\t\t<div class=\"question-content\"><div><span class=\"watupro_num\">1. <\/span>The same amount of perfluoroalkyl acids - perfluorohexanoic acid (PFHxA) and perfluorooctanoic acid (PFOA) was spiked to the different water matrices: ground water, surface water and sea water.<br>\r\n<br>\r\nConcentration of the PFHxA and PFOA was measured from these matrixes using LC-MS. Please indicate the correct answers based on the experimental data:<br>\r\n<table style=\"border-collapse: collapse;\" border=\"1\" cellspacing=\"0\" cellpadding=\"4\"><tbody><tr><td style=\"border: 1px solid black;\" valign=\"top\"><p style=\"text-align: center;\"><strong>Compound<\/strong><\/p><\/td><td style=\"border: 1px solid black; text-align: center; vertical-align: middle;\" colspan=\"3\" valign=\"top\"><strong>\u00a0 \u00a0Matrix: Ground water (n=3) [ng\/L]\u00a0 \u00a0<\/strong><\/td><td style=\"border: 1px solid black; text-align: center; vertical-align: middle;\" colspan=\"3\" valign=\"top\"><strong>\u00a0 \u00a0Matrix: Surface water (n=3) [ng\/L]\u00a0 \u00a0<\/strong><\/td><td style=\"border: 1px solid black;\" colspan=\"3\" valign=\"top\"><p style=\"text-align: center;\"><strong>\u00a0 \u00a0Matrix: Sea water (n=3) [ng\/L]\u00a0 \u00a0<\/strong><\/p><\/td><\/tr><tr><td style=\"border: 1px solid black;\" valign=\"top\"><p style=\"text-align: center;\"><strong>PFHxA<\/strong><\/p><\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">11.4<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">11.2<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">12.6<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">18.9<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">17.7<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">17.5<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">15.7<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">15.6<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">15.4<\/td><\/tr><tr><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\"><strong>PFOA<\/strong><\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">9.1<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">10.0<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">9.7<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">10.9<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">10.9<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">11.0<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">10.4<\/td><td style=\"text-align: center; border: 1px solid black;\" valign=\"top\">10.8<\/td><td style=\"border: 1px solid black;\" valign=\"top\"><p style=\"text-align: center;\">11.0<\/p><\/td><\/tr><\/tbody><\/table><\/div><input type=\"hidden\" name=\"question_id[]\" id=\"qID_1\" value=\"107\" class=\"watupro-question-id\"><input type=\"hidden\" id=\"answerType107\" class=\"answerTypeCnt1\" value=\"checkbox\"><!-- end question-content--><\/div><div class=\"question-choices watupro-choices-columns \" id=\"questionChoices107\"><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-107[]\" id=\"answer-id-279\" class=\"answer   answerof-107 \" value=\"279\"><label for=\"answer-id-279\" id=\"answer-label-279\" class=\" answer\"><span><p>The change in the sample matrix influences PFHxA the most. PFOA remains more robust for the changes in the sample matrix.<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-107[]\" id=\"answer-id-278\" class=\"answer   answerof-107 \" value=\"278\"><label for=\"answer-id-278\" id=\"answer-label-278\" class=\" answer\"><span><p>Used LC-MS method is reliable measuring both analytes in three studies matrices.<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-107[]\" id=\"answer-id-277\" class=\"answer   answerof-107 \" value=\"277\"><label for=\"answer-id-277\" id=\"answer-label-277\" class=\" answer\"><span><p>Both analytes are robust for the changes in the sample matrix.<\/p><\/span><\/label><\/div><!-- end question-choices--><\/div><!-- end questionWrap--><\/div>\t\t\t\t\t<div style=\"display:none;\" id=\"liveResult-1\">\t\t   \n\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/sisu.ut.ee\/wp-content\/plugins\/watupro\/img\/loading.gif\" width=\"16\" height=\"16\" alt=\"Loading...\" title=\"Loading...\">\u00a0Loading...\t\t\t\t\t<\/div>\t\n\t\t\t<\/div> \n\t   \t<p id=\"liveResBtn-107\" style=\"clear:both;\"><input type=\"button\" id=\"liveResultBtn107\" class=\"liveResultBtn1 watupro-live-result-btn\" value=\"See Answer\" onclick=\"WatuPRO.liveResult(107, 1);\"><\/p>\n\t   <div class=\"watu-question \" id=\"question-2\" style=\";\"><div id=\"questionWrap-2\" class=\"   watupro-question-id-108\">\n\t\t\t<div class=\"question-content\"><div><span class=\"watupro_num\">2. <\/span>Indicate correct statements.<\/div><input type=\"hidden\" name=\"question_id[]\" id=\"qID_2\" value=\"108\" class=\"watupro-question-id\"><input type=\"hidden\" id=\"answerType108\" class=\"answerTypeCnt2\" value=\"checkbox\"><!-- end question-content--><\/div><div class=\"question-choices watupro-choices-columns \" id=\"questionChoices108\"><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-108[]\" id=\"answer-id-280\" class=\"answer   answerof-108 \" value=\"280\"><label for=\"answer-id-280\" id=\"answer-label-280\" class=\" answer\"><span><p>Different matrices can lead to different matrix effects and therefore influence also other validation parameters such as trueness, LoQ\/LoD.<strong><\/strong><\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-108[]\" id=\"answer-id-282\" class=\"answer   answerof-108 \" value=\"282\"><label for=\"answer-id-282\" id=\"answer-label-282\" class=\" answer\"><span><p>Sample preparation removes any interferences caused by different sample matrices.\u00a0<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-108[]\" id=\"answer-id-281\" class=\"answer   answerof-108 \" value=\"281\"><label for=\"answer-id-281\" id=\"answer-label-281\" class=\" answer\"><span><p>Visually the same matrix contributes to the matrix effects the same way: an apple is always an apple and human blood plasma is always human blood plasma.<\/p><\/span><\/label><\/div><!-- end question-choices--><\/div><!-- end questionWrap--><\/div>\t\t\t\t\t<div style=\"display:none;\" id=\"liveResult-2\">\t\t   \n\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/sisu.ut.ee\/wp-content\/plugins\/watupro\/img\/loading.gif\" width=\"16\" height=\"16\" alt=\"Loading...\" title=\"Loading...\">\u00a0Loading...\t\t\t\t\t<\/div>\t\n\t\t\t<\/div> \n\t   \t<p id=\"liveResBtn-108\" style=\"clear:both;\"><input type=\"button\" id=\"liveResultBtn108\" class=\"liveResultBtn2 watupro-live-result-btn\" value=\"See Answer\" onclick=\"WatuPRO.liveResult(108, 2);\"><\/p>\n\t   <div class=\"watu-question \" id=\"question-3\" style=\";\"><div id=\"questionWrap-3\" class=\"   watupro-question-id-109\">\n\t\t\t<div class=\"question-content\"><div><span class=\"watupro_num\">3. <\/span>Indicate the validation parameters that are influenced by the differences in visually the same matrix:<\/div><input type=\"hidden\" name=\"question_id[]\" id=\"qID_3\" value=\"109\" class=\"watupro-question-id\"><input type=\"hidden\" id=\"answerType109\" class=\"answerTypeCnt3\" value=\"checkbox\"><!-- end question-content--><\/div><div class=\"question-choices watupro-choices-columns \" id=\"questionChoices109\"><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-109[]\" id=\"answer-id-285\" class=\"answer   answerof-109 \" value=\"285\"><label for=\"answer-id-285\" id=\"answer-label-285\" class=\" answer\"><span><p>Recovery (trueness)<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-109[]\" id=\"answer-id-288\" class=\"answer   answerof-109 \" value=\"288\"><label for=\"answer-id-288\" id=\"answer-label-288\" class=\" answer\"><span><p>Linearity<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-109[]\" id=\"answer-id-283\" class=\"answer   answerof-109 \" value=\"283\"><label for=\"answer-id-283\" id=\"answer-label-283\" class=\" answer\"><span><p>Selectivity<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-109[]\" id=\"answer-id-287\" class=\"answer   answerof-109 \" value=\"287\"><label for=\"answer-id-287\" id=\"answer-label-287\" class=\" answer\"><span><p>Limit of detection\/Limit of quantification<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-109[]\" id=\"answer-id-284\" class=\"answer   answerof-109 \" value=\"284\"><label for=\"answer-id-284\" id=\"answer-label-284\" class=\" answer\"><span><p>Stability<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-109[]\" id=\"answer-id-286\" class=\"answer   answerof-109 \" value=\"286\"><label for=\"answer-id-286\" id=\"answer-label-286\" class=\" answer\"><span><p>Ionization suppression\/enhancement<\/p><\/span><\/label><\/div><!-- end question-choices--><\/div><!-- end questionWrap--><\/div>\t\t\t\t\t<div style=\"display:none;\" id=\"liveResult-3\">\t\t   \n\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/sisu.ut.ee\/wp-content\/plugins\/watupro\/img\/loading.gif\" width=\"16\" height=\"16\" alt=\"Loading...\" title=\"Loading...\">\u00a0Loading...\t\t\t\t\t<\/div>\t\n\t\t\t<\/div> \n\t   \t<p id=\"liveResBtn-109\" style=\"clear:both;\"><input type=\"button\" id=\"liveResultBtn109\" class=\"liveResultBtn3 watupro-live-result-btn\" value=\"See Answer\" onclick=\"WatuPRO.liveResult(109, 3);\"><\/p>\n\t   <div class=\"watu-question \" id=\"question-4\" style=\";\"><div id=\"questionWrap-4\" class=\"   watupro-question-id-166\">\n\t\t\t<div class=\"question-content\"><p><span class=\"watupro_num\">4. <\/span>A LC-MS assay was developed and validated in the laboratory to quantify penicillin G (antibiotic) from human blood serum.<\/p>\n<p>Serum samples were thawed at room temperature and 50 \u00b5L of serum was transferred into a 0.2 mL tube. For a serum an extraction 50 \u00b5L of acetonitrile (containing piperacillin as internal standard (I.S.) in a concentration of 10 \u00b5g\/mL) was added. After vigorous shaking in a Vortex mixer for 2 min, the sample was centrifuged at 13 000 rpm for five minutes until the supernatant (approximately 75 \u00b5L) was separated. The supernatant was then filtrated through a 0.22 \u00b5m filter and transferred into ultra high performance liquid chromatography (UHPLC) autosampler vials using inserts.<\/p>\n<p>Samples were analysed using UHPLC coupled with triple quadrupole. ESI source was used in the positive ionization mode.<\/p>\n<p>When study samples arrived to the laboratory for the analysis, then approximately 20% of them were visually haemolysed (serum contains red blood cells) and 10% hyperlipidemic (serum contains abnormal amount of lipids).<\/p>\n<p>Please evaluate the possible solutions to test matrix robustness during the validation.<\/p>\n<p>\u00a0<\/p>\n<table style=\"border-collapse: collapse; border: 1px solid black;\" border=\"1\" width=\"70%\" cellspacing=\"0\" cellpadding=\"4\">\n<tbody>\n<tr>\n<td style=\"border: 1px solid black;\" width=\"60%\"><strong>Possible solution<\/strong><\/td>\n<td style=\"border: 1px solid black;\" width=\"40%\"><strong>Evaluation<\/strong><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Blood cell content does not affect the assay performance and can be left out during the matrix robustness\/selectivity studies<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_166_1\" class=\"watupro-gap answer answerof-166\"><option value=\"\"><\/option><option value=\"Unusuable\">Unusuable<\/option><option value=\"Average\">Average<\/option><option value=\"Very good\">Very good<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Lipid concentration has low effect on the assay performance and can be left out during the matrix robustness\/selectivity studies<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_166_2\" class=\"watupro-gap answer answerof-166\"><option value=\"\"><\/option><option value=\"Unusuable\">Unusuable<\/option><option value=\"Average\">Average<\/option><option value=\"Very good\">Very good<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Use different blood serums from 6 different sources to evaluate the matrix robustness.<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_166_3\" class=\"watupro-gap answer answerof-166\"><option value=\"\"><\/option><option value=\"Average\">Average<\/option><option value=\"Very good\">Very good<\/option><option value=\"Unusuable\">Unusuable<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Evaluate the effect of haemolysis with spiking the blood serum with different amount (in the range of 0-10%) of whole blood and evaluate the effect of blood content for the assay performance<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_166_4\" class=\"watupro-gap answer answerof-166\"><option value=\"\"><\/option><option value=\"Average\">Average<\/option><option value=\"Very good\">Very good<\/option><option value=\"Unusuable\">Unusuable<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Use matrix with \u2265 240 mg\/dL (total) cholesterol available from a commercial source to evaluate the effect of hyperlipidemic blood serum<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_166_5\" class=\"watupro-gap answer answerof-166\"><option value=\"\"><\/option><option value=\"Average\">Average<\/option><option value=\"Unusuable\">Unusuable<\/option><option value=\"Very good\">Very good<\/option><\/select><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<input type=\"hidden\" name=\"question_id[]\" id=\"qID_4\" value=\"166\" class=\"watupro-question-id\"><input type=\"hidden\" id=\"answerType166\" class=\"answerTypeCnt4\" value=\"gaps\"><!-- end question-content--><\/div><!-- end questionWrap--><\/div>\t\t\t\t\t<div style=\"display:none;\" id=\"liveResult-4\">\t\t   \n\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/sisu.ut.ee\/wp-content\/plugins\/watupro\/img\/loading.gif\" width=\"16\" height=\"16\" alt=\"Loading...\" title=\"Loading...\">\u00a0Loading...\t\t\t\t\t<\/div>\t\n\t\t\t<\/div> \n\t   \t<p id=\"liveResBtn-166\" style=\"clear:both;\"><input type=\"button\" id=\"liveResultBtn166\" class=\"liveResultBtn4 watupro-live-result-btn\" value=\"See Answer\" onclick=\"WatuPRO.liveResult(166, 4);\"><\/p>\n\t   <div style=\"display:none\" id=\"question-5\">\n\t<div class=\"question-content\">\n\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/sisu.ut.ee\/wp-content\/plugins\/watupro\/img\/loading.gif\" width=\"16\" height=\"16\" alt=\"Loading...\" title=\"Loading...\">\u00a0Loading...\t<\/div>\n<\/div>\n\n<br>\n\t\n\t\t\t<div class=\"watupro_buttons flex \" id=\"watuPROButtons26\">\n\t\t  <div id=\"prev-question\" style=\"display:none;\"><input type=\"button\" value=\"&lt; 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