title: Determination of process-related impurities in N-acetylglucosamine prepared by chemical and enzymatic methods: structural elucidation and quantification.
abstract: β-N-acetylglucosamine (β-AG) is a monosaccharide distributed widely in living organisms with various pivotal roles. The presence of particulates and impurities can affect the safety and efficacy of a product for its intended duration of use. Thus, the current study was carried out to identify and quantify the potentially-harmful process related impurities; namely α-N,6-diacetylglucosamine (α-DAG) and α-N-acetylglucosamine (α-AG), derived from the chemical and enzymatic synthesis of β-AG. The impurities were characterized using a high resolution mass spectrometry, a nuclear magnetic resonance spectroscopy, and liquid chromatography-tandem mass spectrometry (LC/MS/MS). The developed method showed a good linearity (R (2) ≥ 0.998), satisfactory precision (≤6.1 % relative standard deviation), intra- and inter-day accuracy (88.20-97.50 %), extraction recovery (89.30-110.50 %), matrix effect (89.70-105.20 %), and stability (92.70-101.60 %). The method was successfully applied to determine the level of α-DAG that was 3.04 and 0.07 % of the total β-AG, following chemical and enzymatic methods, respectively. It can be concluded that the enzymatic rather than the chemical method is more efficient for the synthesis of β-AG. Characterization of impurities heeds the signal for acquiring and evaluating data that establishes biological safety.
authors.affiliation: Amicogen, Inc., 64 Dongburo 1259, Jinsung, Jinju, 660-852, Republic of Korea.
authors.forename: Yi Soo
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Department of Chemistry and Research Institute of Life Science, Gyeongsang National University, Jinju, 660-701, Republic of Korea.
authors.forename: Sung Joong
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Amicogen, Inc., 64 Dongburo 1259, Jinsung, Jinju, 660-852, Republic of Korea.|Department of Chemistry and Research Institute of Life Science, Gyeongsang National University, Jinju, 660-701, Republic of Korea.
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Choi
authors.affiliation: Department of Chemistry and Research Institute of Life Science, Gyeongsang National University, Jinju, 660-701, Republic of Korea.
authors.forename: Yun-Hi
authors.initials: YH
authors.lastname: Kim
authors.affiliation: Department of Chemistry and Research Institute of Life Science, Gyeongsang National University, Jinju, 660-701, Republic of Korea.
authors.forename: Kebede Taye
authors.initials: KT
authors.lastname: Desta
authors.affiliation: Amicogen, Inc., 64 Dongburo 1259, Jinsung, Jinju, 660-852, Republic of Korea.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Piao
authors.affiliation: Amicogen, Inc., 64 Dongburo 1259, Jinsung, Jinju, 660-852, Republic of Korea.
authors.forename: Su-Lim
authors.initials: SL
authors.lastname: Choi
authors.affiliation: Department of Chemistry and Nano Science, Global Top5 Program, Ewha Womans University, Seoul, 120-750, Republic of Korea.
authors.forename: Sang-Jip
authors.initials: SJ
authors.lastname: Nam
authors.affiliation: Department of Pharmacy, Sunchon National University, Suncheon, 540-742, Republic of Korea.
authors.forename: Kyung-Yun
authors.initials: KY
authors.lastname: Kang
authors.affiliation: Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Konkuk University, 1 Hwayang-dong, Kwangjin-gu, Seoul, 143-701, Republic of Korea.|Department of Pharmacology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt.
authors.forename: A M
authors.initials: AM
authors.lastname: Abd El-Aty
authors.affiliation: Amicogen, Inc., 64 Dongburo 1259, Jinsung, Jinju, 660-852, Republic of Korea. ycshin@amicogen.com.
authors.forename: Yong Chul
authors.initials: YC
authors.lastname: Shin
authors.affiliation: Department of Chemistry and Research Institute of Life Science, Gyeongsang National University, Jinju, 660-701, Republic of Korea. sshin@gnu.ac.kr.
authors.forename: Sung Chul
authors.initials: SC
authors.lastname: Shin
mesh_terms: D000117:Acetylglucosamine / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D004340:Drug Contamination; D057075:Enzyme Assays / Q000379:methods; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000117:Acetylglucosamine
keywords: Chitin; Nuclear magnetic resonance; Process-related impurity; Tandem mass spectrometry; β-N-acetylglucosamine