{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T18:21:02Z","timestamp":1781115662168,"version":"3.54.1"},"reference-count":7,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,6,8]],"date-time":"2023-06-08T00:00:00Z","timestamp":1686182400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Science Education and Workforce Development Programs at Oak Ridge National Laboratory"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJNS"],"abstract":"<jats:p>Mucopolysaccharidosis type II (MPS-II, Hunter syndrome, OMIM:30990) is a lysosomal storage disorder (LSD) that results in iduronate 2-sulphatase (I2S) enzyme deficiency. MPS-II was added to the Recommended Uniform Screening Panel (RUSP) in August 2022; thus, there is an increased demand for multiplexing I2S into existing LSD screening assays. After incubation with LSD synthetic substrates, extracts are cleaned using liquid\u2013liquid extraction with ethyl acetate or protein precipitation using acetonitrile (ACN). We investigated cold-induced water ACN phase separation (CIPS) to improve the combination of 6-plex and I2S extracts to create a 7-plex assay, and compared it to room temperature ACN and ethyl acetate liquid\u2013liquid extraction. The extracts were dried and resuspended in the mobile phase, and then analyzed using an optimized 1.9 min injection-to-injection liquid chromatography method coupled with tandem mass spectrometry (LC-MS\/MS). The combination of ACN and CIPS improved the detection for I2S products without significant detriment to other analytes, which is attributable to a more complete coagulation and separation of heme, proteins, and extracted residual salts. Using CIPS for sample cleanup in dried blood spots (DBS) appears to represent a promising and straightforward way of achieving cleaner sample extracts in a new 7-plex LSD screening panel.<\/jats:p>","DOI":"10.3390\/ijns9020032","type":"journal-article","created":{"date-parts":[[2023,6,9]],"date-time":"2023-06-09T02:03:18Z","timestamp":1686276198000},"page":"32","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Multiplexing Iduronate-2-Sulphatase (MPS-II) into a 7-Plex Lysosomal Storage Disorder MS\/MS Assay Using Cold-Induced Phase Separation"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5304-8126","authenticated-orcid":false,"given":"Elya","family":"Courtney","sequence":"first","affiliation":[{"name":"Centers for Disease Control and Prevention, Atlanta, GA 30341, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1632-5777","authenticated-orcid":false,"given":"C. Austin","family":"Pickens","sequence":"additional","affiliation":[{"name":"Centers for Disease Control and Prevention, Atlanta, GA 30341, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carla","family":"Cuthbert","sequence":"additional","affiliation":[{"name":"Centers for Disease Control and Prevention, Atlanta, GA 30341, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8660-8532","authenticated-orcid":false,"given":"Konstantinos","family":"Petritis","sequence":"additional","affiliation":[{"name":"Centers for Disease Control and Prevention, Atlanta, GA 30341, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"D\u2019Avanzo, F., Rigon, L., Zanetti, A., and Tomanin, R. (2020). Mucopolysaccharidosis Type II: One Hundred Years of Research, Diagnosis, and Treatment. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21041258"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1016\/j.ymgme.2016.05.015","article-title":"Pilot study of newborn screening for six lysosomal storage diseases using Tandem Mass Spectrometry","volume":"118","author":"Elliott","year":"2016","journal-title":"Mol. Genet. Metab."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.jpeds.2019.07.053","article-title":"Population-Based Newborn Screening for Mucopolysaccharidosis Type II in Illinois: The First Year Experience","volume":"214","author":"Burton","year":"2019","journal-title":"J. Pediatr."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1016\/j.jpeds.2017.06.048","article-title":"Newborn Screening for Lysosomal Storage Disorders in Illinois: The Initial 15-Month Experience","volume":"190","author":"Burton","year":"2017","journal-title":"J. Pediatr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1373\/clinchem.2012.189936","article-title":"High-Throughput Assay of 9 Lysosomal Enzymes for Newborn Screening","volume":"59","author":"Spacil","year":"2013","journal-title":"Clin. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.chroma.2017.05.045","article-title":"Cold-induced aqueous ACN phase separation: A salt-free way to begin quick, easy, cheap, effective, rugged, safe","volume":"1506","author":"Shao","year":"2017","journal-title":"J. Chromatogr. A"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/j.jpeds.2019.09.036","article-title":"Newborn Screening for Mucopolysaccharidoses: Results of a Pilot Study with 100,000 Dried Blood Spots","volume":"216","author":"Scott","year":"2020","journal-title":"J. Pediatr."}],"container-title":["International Journal of Neonatal Screening"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2409-515X\/9\/2\/32\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:51:11Z","timestamp":1760125871000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2409-515X\/9\/2\/32"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,8]]},"references-count":7,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["ijns9020032"],"URL":"https:\/\/doi.org\/10.3390\/ijns9020032","relation":{},"ISSN":["2409-515X"],"issn-type":[{"value":"2409-515X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,8]]}}}