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Existing methods are susceptible to ion suppression in complex serum matrices and therefore usually require laborious sample preparation and time-consuming chromatographic separation, limiting their application in high-throughput screening and rapid testing. In this study, we developed an integrated purification-enrichment-derivatization paper chip, termed iPED chip, which integrates diatomaceous earth-assisted online cleanup at the tail end, thermal enrichment at the paper tip, and in situ hydroxylamine derivatization. A programmable induction heating device was also developed to provide a localized, controllable, and noncontact thermal field for the paper reaction zone, thereby promoting analyte enrichment and reaction. Coupled with tandem mass spectrometry, the iPED chip-MS\/MS method enabled rapid simultaneous quantification of testosterone (T), androstenedione (4-AD), dehydroepiandrosterone (DHEA), and 17\u03b1-hydroxyprogesterone (17-OHP) in serum within 20 min. Method characterization demonstrated effective enrichment of the target hormones, broad linearity over 0.078\u201340 \u03bcg\/L in spiked calf serum, low limits of detection of 0.042\u20130.093 \u03bcg\/L, recoveries of 95\u2013106%, and RSDs below 10%. In spiked hormone-depleted human serum samples at clinically relevant levels, the results agreed well with those obtained by conventional LC\u2013MS\/MS, supporting the reliability of iPED chip-MS\/MS and its potential for rapid clinical testing and large-scale biofluid screening.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c04260","type":"journal-article","created":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T14:01:36Z","timestamp":1786802496000},"page":"27077-27087","source":"Crossref","is-referenced-by-count":0,"title":["A Heating-Assisted\nIntegrated Purification-Enrichment-Derivatization\nPaper Chip for Rapid MS\/MS Quantification of Serum Steroid Hormones"],"prefix":"10.1021","volume":"98","author":[{"given":"Can","family":"Tan","sequence":"first","affiliation":[{"name":"Shenzhen Campus of Sun Yat-sen University , , , ,","place":["Shenzhen, Guangdong, P. R. 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China, 518107"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuan","family":"Yu","sequence":"additional","affiliation":[{"name":"Shenzhen Campus of Sun Yat-sen University , , , ,","place":["Shenzhen, Guangdong, P. R. China, 518107"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenqing","family":"Cai","sequence":"additional","affiliation":[{"name":"Shenzhen Bioeasy Biotechnology Co., Ltd. , , ,","place":["Shenzhen, Guangdong, P. R. China, 518126"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Le","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Campus of Sun Yat-sen University , , , ,","place":["Shenzhen, Guangdong, P. R. China, 518107"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuo","family":"Lei","sequence":"additional","affiliation":[{"name":"Shenzhen Campus of Sun Yat-sen University , , , ,","place":["Shenzhen, Guangdong, P. R. China, 518107"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3204-8717","authenticated-orcid":true,"given":"Yiyong","family":"Yan","sequence":"additional","affiliation":[{"name":"Shenzhen Polytechnic University , , , ,","place":["Shenzhen, Guangdong, P. R. China, 518055"]},{"name":"Shenzhen Bioeasy Biotechnology Co., Ltd. , , ,","place":["Shenzhen, Guangdong, P. R. China, 518126"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9895-8679","authenticated-orcid":true,"given":"Jianhua","family":"Zhou","sequence":"additional","affiliation":[{"name":"Shenzhen Campus of Sun Yat-sen University , , , ,","place":["Shenzhen, Guangdong, P. R. 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Bioprospect."},{"key":"2026091511080328100_cit37","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1016\/j.jmsacl.2025.10.003","article-title":"Measurement of Twenty-One\nSerum Steroid Profiles by UPLC-MS\/MS for\nthe Diagnosis and Monitoring of Congenital Adrenal Hyperplasia","volume":"38","author":"Lee","year":"2025","journal-title":"J. Mass Spectrom. Adv. Clin. Lab"},{"issue":"3","key":"2026091511080328100_cit38","doi-asserted-by":"publisher","first-page":"537","DOI":"10.1007\/s13361-011-0308-4","article-title":"Structure\nElucidation of the Diagnostic Product Ion at m\/z 97 Derived from Androst-4-En-3-One-Based\nSteroids by ESI-CID and IRMPD Spectroscopy","volume":"23","author":"Thevis","year":"2012","journal-title":"J. Am. Soc. 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Chem."},{"issue":"4","key":"2026091511080328100_cit41","doi-asserted-by":"publisher","first-page":"734","DOI":"10.1016\/j.rbmo.2020.07.013","article-title":"The Predictive\nValue of Total Testosterone Alone for Clinical Hyperandrogenism in\nPolycystic Ovary Syndrome","volume":"41","author":"Yang","year":"2020","journal-title":"Reprod. BioMed. Online"},{"issue":"4","key":"2026091511080328100_cit42","doi-asserted-by":"publisher","first-page":"580","DOI":"10.1194\/jlr.D119000591","article-title":"An LC\/MS\/MS Method for Analyzing\nthe Steroid Metabolome with High Accuracy and from Small Serum Samples","volume":"61","author":"Yuan","year":"2020","journal-title":"J. Lipid Res."},{"issue":"6","key":"2026091511080328100_cit43","doi-asserted-by":"publisher","first-page":"bvaa047","DOI":"10.1210\/jendso\/bvaa047","article-title":"An Ultrasensitive\nRoutine LC-MS\/MS Method for Estradiol and Estrone in the Clinically\nRelevant Sub-Picomolar Range","volume":"4","author":"Bertelsen","year":"2020","journal-title":"J. Endocr. Soc."},{"issue":"31","key":"2026091511080328100_cit44","doi-asserted-by":"publisher","first-page":"3303","DOI":"10.1016\/j.jchromb.2010.10.012","article-title":"Investigation\nof\nEndogenous Blood Plasma Phospholipids, Cholesterol and Glycerides\nThat Contribute to Matrix Effects in Bioanalysis by Liquid Chromatography\/Mass\nSpectrometry","volume":"878","author":"Ismaiel","year":"2010","journal-title":"J. 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