
{"id":2558,"date":"2022-05-02T00:00:00","date_gmt":"2022-05-02T00:00:00","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/analyteguru\/determining-nitrite-in-pharmaceutical-products-using-ion\/"},"modified":"2024-04-16T18:37:58","modified_gmt":"2024-04-16T18:37:58","slug":"determining-nitrite-in-pharmaceutical-products-using-ion","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/analyteguru\/determining-nitrite-in-pharmaceutical-products-using-ion\/","title":{"rendered":"Determining Nitrite in Pharmaceutical Products Using Ion Chromatography and UV Detection"},"content":{"rendered":"<div class=\"lia-message-body-content\">\n<h4 id=\"toc-hId--1398085222\"><strong><span style=\"color: #000000\"><em>Editor&#8217;s Note: Dr. Jeffrey Rohrer, Director of Applications Development, contributed to this article.<\/em><\/span><\/strong><\/h4>\n<p>N-nitrosodimethylamine (NDMA) is a highly hepatotoxic compound and known carcinogen. In recent years, several drugs, including losartan, ranitidine, and metformin, have been recalled by the U.S. Food and Drug Administration (FDA) due to the presence of NDMA. Pharmaceutical manufacturing processes that use or generate nitrite can lead to NDMA formation if a secondary or tertiary amine is present. Monitoring nitrite and amine levels during drug manufacturing is, therefore, important to minimize development of NDMA in pharmaceutical products.<\/p>\n<h3 id=\"toc-hId-1154321813\"><strong><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-10795 alignright\" src=\"http:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Determining-Nitrite-300x152.jpg\" alt=\"\" width=\"499\" height=\"253\" srcset=\"https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Determining-Nitrite-300x152.jpg 300w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Determining-Nitrite-768x390.jpg 768w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Determining-Nitrite.jpg 1000w\" sizes=\"auto, (max-width: 499px) 100vw, 499px\" \/>Overcoming chloride ion interference in nitrite determination<\/strong><\/h3>\n<p><a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/industrial\/chromatography\/ion-chromatography-ic.html\" target=\"_blank\" rel=\"noopener\">Ion chromatography (IC)<\/a> is widely used within the pharmaceutical industry for the determination of ionic analytes in products containing non-ionic components. IC-based methods are included in several U. S. Pharmacopeia (USP) monographs and have been applied across many pharmaceutical manufacturing applications, including the determination of active ingredients, degradation products and impurities.<\/p>\n<p>The determination of common anions, including nitrite and nitrate, is usually achieved by coupling anion-exchange separation with suppressed conductivity detection. However, in some drug products, chloride is present as the counter-ion at very high concentrations. This poses a challenge when quantifying low \u03bcg\/L concentrations of nitrite using suppressed conductivity detection, as the large chloride signal may mask the small nitrite peak and make integration difficult.<\/p>\n<p>To overcome this challenge, UV absorption can be used as an alternative detection technique. Detection by UV absorption supports sensitive and selective quantification of nitrite without interference from chloride. Additionally, advanced column chemistries can also be employed to improve the resolution of nitrite and chloride . High-capacity, high-resolution anion-exchange columns, such as the <a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/083217\" target=\"_blank\" rel=\"nofollow noopener\">Thermo Scientific\u2122 Dionex\u2122 IonPac\u2122 AS19-4 \u03bcm column<\/a>, support efficient separation of nitrite even in the presence of large quantities of <a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/industrial\/pharma-biopharma\/pharma-biopharma-learning-center\/pharmaceutical-qa-qc-information\/counter-ion-analysis-information.html\" target=\"_self\" rel=\"nofollow noopener\">counter-ions<\/a> such as chloride.<\/p>\n<h3 id=\"toc-hId-2041825494\"><strong>Effective separation and detection of nitrite in pharmaceutical products<\/strong><\/h3>\n<p>We demonstrated the effectiveness of IC for nitrite analysis using UV detection in a recent <a href=\"https:\/\/assets.thermofisher.com\/TFS-Assets\/CMD\/Application-Notes\/an-73987-ic-nitrite-pharmaceuticals-an73987-en.pdf\" target=\"_blank\" rel=\"nofollow noopener\">application note<\/a>. Using a Dionex IonPac AS19-4 \u03bcm column on a <a href=\"https:\/\/www.thermofisher.com\/uk\/en\/home\/industrial\/chromatography\/ion-chromatography-ic\/ion-chromatography-systems\/modular-ic-systems.html\" target=\"_blank\" rel=\"nofollow noopener\">Thermo Scientific\u2122 Dionex\u2122 ICS-6000 HPIC\u2122 system<\/a>, we analyzed nitrite content in seven pharmaceutical samples (two drug substances and five drug products). For the purposes of comparison, analyte detection was performed by suppressed conductivity and UV absorbance at 210 nm.<\/p>\n<p>Using a potassium hydroxide gradient, the Dionex IonPac AS19-4 \u03bcm column achieved efficient separation of a mixed reference solution of nitrite and six other common anions (Figure 1). While all seven ions were detected by suppressed conductivity, only three (nitrite, nitrate, and bromide) had UV absorbance at 210 nm, resulting in a cleaner chromatogram for subsequent studies. The limit of detection, based on the signal-to-noise (S\/N) ratio, was determined to be 0.918 \u03bcg\/L (corresponding to 0.918 ppm). Excellent linearity (r<sup>2<\/sup>&gt;0.999) was observed over the calibration range of 5 to 500 \u03bcg\/L, highlighting the suitability of the method.<\/p>\n<h3 id=\"toc-hId--1365638121\"><strong>Using ion chromatography to determine nitrite in pharmaceutical samples<\/strong><\/h3>\n<p>This method was used to determine levels of nitrite in two drug substances (losartan potassium and metformin hydrochloride) and five drug products (including two brands of diphenhydramine hydrochloride, metformin hydrochloride, losartan potassium, and ranitidine hydrochloride). Different lots or preparations of three of the drug products (losartan, metformin, and ranitidine) had been recalled by the FDA due to the presence of NDMA.<\/p>\n<p>Nitrite was detected in all samples except the losartan potassium. As expected, the high chloride concentration in the hydrochloride drug products strongly affected the ability to detect nitrite by suppressed conductivity. However, with UV detection at 210 nm, chloride did not interfere with nitrite quantification, permitting cleaner peak integration.<\/p>\n<p>Method accuracy was evaluated by determining recoveries of nitrite spiked into each sample at 10 \u03bcg\/L. Nitrite recoveries for the seven samples ranged from 96 to 101%. Method precision was determined using injections of the 50 \u03bcg\/L nitrite calibration standard on three separate days. The peak area precision was 0.56%, and retention time precision was 0.10%.<\/p>\n<h3 id=\"toc-hId--478134440\"><strong>Sensitive nitrite determination for pharmaceutical manufacturing processes<\/strong><\/h3>\n<p>To support the production of safe and effective pharmaceuticals, reliable methods for determining nitrite in drug substances and products are essential. The IC method highlighted here used a high-capacity anion-exchange column and UV absorption detection to overcome chloride ion interference challenges, resulting in sensitive and accurate nitrite determination.<\/p>\n<p>Read more about this method for <a href=\"https:\/\/assets.thermofisher.com\/TFS-Assets\/CMD\/Application-Notes\/an-73987-ic-nitrite-pharmaceuticals-an73987-en.pdf\" target=\"_blank\" rel=\"nofollow noopener\">the determination of nitrite in pharmaceuticals<\/a>.<\/p>\n<p><span class=\"lia-inline-image-display-wrapper lia-image-align-center\" style=\"width: 624px\"><span class=\"lia-inline-image-caption\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-10794\" src=\"http:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Figure-1.png\" alt=\"\" width=\"800\" height=\"377\" srcset=\"https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Figure-1.png 624w, https:\/\/admin.acceleratingscience.com\/analyteguru\/wp-content\/uploads\/sites\/25\/2023\/06\/050222-Figure-1-300x141.png 300w\" sizes=\"auto, (max-width: 800px) 100vw, 800px\" \/>Figure 1. Separation of seven common anions using a Dionex IonPac AS19-4\u03bcm column.<\/span><\/span><\/p>\n<h4 id=\"toc-hId--1398085222\"><strong>On LinkedIn? Visit our\u00a0<a href=\"https:\/\/www.linkedin.com\/showcase\/chromatography-&amp;-mass-spectrometry\/posts\/?feedView=all\" target=\"_blank\" rel=\"noreferrer noopener\" data-uw-rm-brl=\"exc\" aria-label=\"LinkedIn page - opens in new tab\" data-uw-rm-ext-link=\"\">LinkedIn page<\/a> #IonChromatography<\/strong><\/h4>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>To support the production of safe and effective pharmaceuticals, reliable methods for determining nitrite in drug substances and products are essential.<\/p>\n","protected":false},"author":1327,"featured_media":2560,"comment_status":"open","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":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[17,28],"tags":[20,31],"division":[],"class_list":{"0":"post-2558","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-ion-chromatography","8":"category-pharmaceutical","9":"tag-ion-chromatography","10":"tag-pharmaceutical","11":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- 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