title: Sample injection strategy to increase throughput in counter-current chromatography: Case study of Honokiol purification.
abstract: Counter-current chromatography (CCC) has been widely used as a preparative separation method to purify natural products from plant extracts and fermentation broths. Traditionally, throughput optimization in CCC has focused on sample concentration and sample volume. In this paper sample injection was considered as consisting of three variables: injection flow rate, post-injection flow rate and sample solvent. The effects of these parameters were studied using a honokiol purification from a Magnolia officinalis bark extract as a case study aiming to achieve the highest throughput/yield ratio for greater than 99% purity of this potential anti-cancer drug obtained for submission to the Chinese FDA. An injection method was established that increased the throughput of honokiol by 46.5% (from 3.05g/h to 4.47g/h), and decreased the solvent consumption of mobile phase and stationary phase per gram of honokiol by 40.0% (from 0.68L/g to 0.41L/g) and 48.4% (from 0.40L/g to 0.21L/g) respectively. These results show the importance of understanding the whole injection process when optimizing a given CCC separation.
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Centre for Bio Therapy, Chengdu 610041, China.
authors.forename: Aihua
authors.initials: A
authors.lastname: Peng
authors.affiliation: Advanced Bioprocessing Centre, College of Engineering, Design and Physical Sciences, Brunel University London, Uxbridge, UB8 3PH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Hewitson
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Centre for Bio Therapy, Chengdu 610041, China.
authors.forename: Haoyu
authors.initials: H
authors.lastname: Ye
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences, School of Medicine, Tsinghua University and Collaborative Innovation Centre for Biotherapy, Beijing 100084, China.
authors.forename: Liansuo
authors.initials: L
authors.lastname: Zu
authors.affiliation: Advanced Bioprocessing Centre, College of Engineering, Design and Physical Sciences, Brunel University London, Uxbridge, UB8 3PH, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Garrard
authors.affiliation: Advanced Bioprocessing Centre, College of Engineering, Design and Physical Sciences, Brunel University London, Uxbridge, UB8 3PH, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Sutherland
authors.affiliation: State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Centre for Bio Therapy, Chengdu 610041, China. Electronic address: chenlijuan125@163.com.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Chen
authors.affiliation: Advanced Bioprocessing Centre, College of Engineering, Design and Physical Sciences, Brunel University London, Uxbridge, UB8 3PH, UK. Electronic address: svetlana.ignatova@brunel.ac.uk.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Ignatova
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification*; D001713:Biphenyl Compounds / Q000302:isolation & purification*; D003377:Countercurrent Distribution / Q000379:methods*; D017705:Lignans / Q000302:isolation & purification*; D031566:Magnolia / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry; D012997:Solvents; D000498:Allyl Compounds; D010636:Phenols
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D001713:Biphenyl Compounds; D017705:Lignans; D010936:Plant Extracts; D012997:Solvents; C005499:honokiol; D000498:Allyl Compounds; D010636:Phenols
keywords: CCC; Counter-current chromatography; Honokiol; Magnolol; Sample injection; Sample loading; Throughput