Immunoaffinity trapping of urinary human chorionic gonadotropin and its high-performance liquid chromatographic-mass spectrometric confirmation |
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| Affiliation: | 1. Soils Department, Federal University of Santa Maria (Universidade Federal de Santa Maria), 97105-900 Santa Maria, RS, Brazil;2. Agronomy Department, University of Santa Catarina State, 88520-000 Lages, SC, Brazil,;3. University of West Santa Catarina, 89620-000 Campos Novos, SC, Brazil;1. College of Land and Environment, Shenyang Agricultural University, Shenyang 110866, China;2. University of Wisconsin–Madison, Department of Soil Science, FD Hole Soils Lab, 1525 Observatory Drive, Madison, WI 53706, USA;1. Department of Reproductive Medicine, Xiamen Maternity and Child Care Hospital, Xiamen, Fujian 361005, China;2. Reproductive Medicine Center, Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong 264000, China;3. Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Chaoyang, Beijing 100026, China;1. College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, PR China;2. National Beef Cattle Improvement Center of Northwest A&F University, Yangling 712100, China;3. College of Animal Science and Technology, Huazhaong Agricultural University, Wuhan, Hubei 430070, China;4. College of Bio-medical engineering Chongqing University Chongqing, China;5. Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, UT 84321, USA |
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| Abstract: | A method has been developed for confirmation of the glycopeptide human chorionic gonadotropin (hCG) in urine. A solid-phase immunoaffinity trapping technique utilizing a monoclonal antibody recognizing both intact hCG and free hCG β-subunits was developed for the extraction of hCG from urine. Recovery of hCG from a urine matrix was essentially quantitative. The hCG was quantitatively eluted with 6 M guanidine hydrochloride, reductively alkylated with vinylpyridine, and subjected to tryptic digestion. The tryptic digest was analysed by HPLC-MS. Ions from three tryptic fragments were monitored with selected ion monitoring to provide specific detection of hCG. The signal observed for a concentration of 25 mIU/ml of hCG could be clearly distinguished from background with a signal-to-noise ratio of 12:1. |
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