{"id":1194,"date":"2024-04-11T14:44:04","date_gmt":"2024-04-11T11:44:04","guid":{"rendered":"https:\/\/sisu.ut.ee\/lcms_method_validation\/self-test-10-3\/"},"modified":"2024-04-11T14:44:04","modified_gmt":"2024-04-11T11:44:04","slug":"self-test-10-3","status":"publish","type":"page","link":"https:\/\/sisu.ut.ee\/lcms_method_validation\/self-test-10-3\/","title":{"rendered":"Self test 10.3"},"content":{"rendered":"<div id=\"watupro_quiz\" class=\"quiz-area single-page-quiz\">\n<p id=\"submittingExam27\" 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-27\" enctype=\"multipart\/form-data\">\n<div class=\"watu-question \" id=\"question-1\" style=\";\"><div id=\"questionWrap-1\" class=\"   watupro-question-id-110\">\n\t\t\t<div class=\"question-content\"><div><span class=\"watupro_num\">1. <\/span>Indicate the most influential changes in method parameters in case the <strong>critical performance parameter<\/strong> is <strong>LoQ<\/strong>.<\/div><input type=\"hidden\" name=\"question_id[]\" id=\"qID_1\" value=\"110\" class=\"watupro-question-id\"><input type=\"hidden\" id=\"answerType110\" class=\"answerTypeCnt1\" value=\"checkbox\"><!-- end question-content--><\/div><div class=\"question-choices watupro-choices-columns \" id=\"questionChoices110\"><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-298\" class=\"answer   answerof-110 \" value=\"298\"><label for=\"answer-id-298\" id=\"answer-label-298\" class=\" answer\"><span><p>Ion source configuration (Nebulizer position)<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-290\" class=\"answer   answerof-110 \" value=\"290\"><label for=\"answer-id-290\" id=\"answer-label-290\" class=\" answer\"><span><p>Mobile phase additives<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-293\" class=\"answer   answerof-110 \" value=\"293\"><label for=\"answer-id-293\" id=\"answer-label-293\" class=\" answer\"><span><p>Eluent flow rate<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-295\" class=\"answer   answerof-110 \" value=\"295\"><label for=\"answer-id-295\" id=\"answer-label-295\" class=\" answer\"><span><p>Sample preparation parameters<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-299\" class=\"answer   answerof-110 \" value=\"299\"><label for=\"answer-id-299\" id=\"answer-label-299\" class=\" answer\"><span><p>Ion source condition (Nebulizer aging, ion source contamination)<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-297\" class=\"answer   answerof-110 \" value=\"297\"><label for=\"answer-id-297\" id=\"answer-label-297\" class=\" answer\"><span><p>Nebulizer gas pressure\/flow rate<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-296\" class=\"answer   answerof-110 \" value=\"296\"><label for=\"answer-id-296\" id=\"answer-label-296\" class=\" answer\"><span><p>Drying gas temperature<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-294\" class=\"answer   answerof-110 \" value=\"294\"><label for=\"answer-id-294\" id=\"answer-label-294\" class=\" answer\"><span><p>Column batch and age<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-292\" class=\"answer   answerof-110 \" value=\"292\"><label for=\"answer-id-292\" id=\"answer-label-292\" class=\" answer\"><span><p>Column temperature<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-291\" class=\"answer   answerof-110 \" value=\"291\"><label for=\"answer-id-291\" id=\"answer-label-291\" class=\" answer\"><span><p>Organic solvent content in the eluent<\/p><\/span><\/label><\/div><div class=\"watupro-question-choice  \" dir=\"auto\"><input type=\"checkbox\" name=\"answer-110[]\" id=\"answer-id-289\" class=\"answer   answerof-110 \" value=\"289\"><label for=\"answer-id-289\" id=\"answer-label-289\" class=\" answer\"><span><p>Mobile phase pH<\/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-110\" style=\"clear:both;\"><input type=\"button\" id=\"liveResultBtn110\" class=\"liveResultBtn1 watupro-live-result-btn\" value=\"See Answer\" onclick=\"WatuPRO.liveResult(110, 1);\"><\/p>\n\t   <div class=\"watu-question \" id=\"question-2\" style=\";\"><div id=\"questionWrap-2\" class=\"   watupro-question-id-167\">\n\t\t\t<div class=\"question-content\"><p><span class=\"watupro_num\">2. <\/span>A LC-MS assay was developed and validated in the laboratory to measure meropenem (antibiotic; p<em>K<\/em><sub>a1<\/sub>=3.47; p<em>K<\/em><sub>a2<\/sub>=9.39) from human blood serum.<\/p>\n<p>Blood serum samples were thawed\u00a0at room temperature and 50 \u00b5L of serum was transferred into 250 \u00b5L tube. For serum sample extraction 50 \u00b5L of methanol (containing ertapenem (p<em>K<\/em><sub>a1<\/sub>=3.37; p<em>K<\/em><sub>a2<\/sub>=9.03) as internal standard (I.S.) in a concentration of 10 \u00b5g\/mL) was added. After vigorous shaking with Vortex mixer for 1 min, the sample was centrifuged at 8\u00a0000 rpm (3500 \u00d7 g) for 10 min and supernatant (approximately 75 \u00b5L) was separated and filtrated through 0.22 \u00b5m filter and transferred into autosampler vial.<\/p>\n<p>Samples were analysed using UHPLC coupled with triple quadrupole. Gradient elution with methanol and 0.1 % formic acid (pH = 2.6) on a flow rate of 0.3 mL\/min was used for chromatographic separation. Samples were chromatographed using C18 column (2.1 \u00d7 100 mm, 1.7 \u00b5m) equipped with C18 pre-column (2.1 \u00d7 5 mm, 1.7 \u00b5m). Column temperature was set to 40 \u2070C. ESI source (allows the change in source configuration) was used in the positive ionization mode. Transitions of the parent ion with\u00a0<em>m\/z<\/em>\u00a0384 [M + H]<sup>+<\/sup>\u00a0to daughter ions\u00a0<em>m\/z\u00a0<\/em>254, 298, 340 were used for meropenem quantification and qualification.<\/p>\n<p>The calibration curves were linear from 0.1 to 200 \u00b5g\/mL in serum. LoQ for serum samples was 0.1 \u00b5g\/mL.<\/p>\n<p>Please evaluate the effect (<strong>high, medium, low<\/strong>) of different method parameters that are affecting method robustness and what can actually change during the analysis of study samples.<\/p>\n<p>\u00a0<\/p>\n<table style=\"border-collapse: collapse; border: 1px solid black;\" border=\"1\" cellspacing=\"0\" cellpadding=\"4\">\n<tbody>\n<tr>\n<td style=\"border: 1px solid black;\" width=\"70%\"><strong>Parameter<\/strong><\/td>\n<td style=\"border: 1px solid black;\" width=\"30%\"><strong>Evaluation of effect<\/strong><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Column temperature<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_1\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Medium\">Medium<\/option><option value=\"High\">High<\/option><option value=\"Low\">Low<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Injection solvent composition<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_2\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Low\">Low<\/option><option value=\"High\">High<\/option><option value=\"Medium\">Medium<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Relative +20 % increase in the eluent flow rate<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_3\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Medium\">Medium<\/option><option value=\"High\">High<\/option><option value=\"Low\">Low<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Ion source configuration (Nebulizer position)<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_4\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Low\">Low<\/option><option value=\"High\">High<\/option><option value=\"Medium\">Medium<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Sample matrix variability (e.g. haemolysis of the serum)<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_5\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Low\">Low<\/option><option value=\"Medium\">Medium<\/option><option value=\"High\">High<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Relative +10 % change in formic acid content in mobile phase<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_6\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Medium\">Medium<\/option><option value=\"Low\">Low<\/option><option value=\"High\">High<\/option><\/select><\/td>\n<\/tr>\n<tr>\n<td style=\"border: 1px solid black;\">Organic solvent content in mobile phase<\/td>\n<td style=\"border: 1px solid black;\"><select name=\"gap_167_7\" class=\"watupro-gap answer answerof-167\"><option value=\"\"><\/option><option value=\"Low\">Low<\/option><option value=\"High\">High<\/option><option value=\"Medium\">Medium<\/option><\/select><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<input type=\"hidden\" name=\"question_id[]\" id=\"qID_2\" value=\"167\" class=\"watupro-question-id\"><input type=\"hidden\" id=\"answerType167\" class=\"answerTypeCnt2\" value=\"gaps\"><!-- end question-content--><\/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-167\" style=\"clear:both;\"><input type=\"button\" id=\"liveResultBtn167\" class=\"liveResultBtn2 watupro-live-result-btn\" value=\"See Answer\" onclick=\"WatuPRO.liveResult(167, 2);\"><\/p>\n\t   <div style=\"display:none\" id=\"question-3\">\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=\"watuPROButtons27\">\n\t\t  <div id=\"prev-question\" style=\"display:none;\"><input type=\"button\" value=\"&lt; 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