Circulating ultrasound-assisted extraction, countercurrent chromatography, and liquid chromatography for the simultaneous extraction, isolation, and analysis of the constituents of Uncaria tomentosa.
A hyphenated automated technique for the online extraction, isolation, analysis, and identification of natural organic compounds was established. Circulating ultrasound-assisted extraction (CUAE) was coupled with countercurrent chromatography (CCC), high performance liquid chromatography (HPLC), and a diode array detector (DAD). This approach was applied to the fractionation and purification of alkaloids from Uncaria tomentosa. A biphasic solvent system of chloroform-methanol-water (6:4:5, v:v:v) was used for the CUAE and CCC separation of compounds from 500 g of U. tomentosa. Two CUAE/CCC/HPLC/DAD modes were established. Either the upper aqueous phase or the lower organic phase of the solvent system could be used as the extraction solvent. The target compounds were extracted by CUAE, and the extract was pumped into a sample loop before being directly injected into the CCC column, or pre-purified using a flash chromatography column before injection. The target compounds were eluted using either the organic or aqueous phase of the solvent system and the fractions were monitored using a UV detector. The target fractions were collected by a sample loop via a six-port valve, and analyzed by HPLC/DAD for purity and structural identification. This system isolated of 8.2mg, 7.4 mg, and 12.9 mg of rhynchophylline, corynoxine, and corynoxine B with HPLC purities of 96.15%, 95.34%, and 95.49%, respectively via the first mode; and isolated 26.6 mg, 24.6 mg, and 45.3mg of rhynchophylline, corynoxine, and corynoxine B with a HPLC purities of 98.22%, 97.18%, and 97.93% via the second mode.
Central Laboratory, Changchun Normal University, No. 677 North Changji Road, Erdao District, Changchun 130032, China.
Yuchi
Y
Zhang
Central Laboratory, Changchun Normal University, No. 677 North Changji Road, Erdao District, Changchun 130032, China. Electronic address: ccsf777@163.com.
Chunming
C
Liu
Central Laboratory, Changchun Normal University, No. 677 North Changji Road, Erdao District, Changchun 130032, China.
Yanjuan
Y
Qi
Central Laboratory, Changchun Normal University, No. 677 North Changji Road, Erdao District, Changchun 130032, China.
Sainan
S
Li
Higher Education Research Institute, Changchun Normal University, No. 677 North Changji Road, Erdao District, Changchun, 130032, China.
Yan
Y
Pan
Traditional Chinese Medicine Academy of Science of Jilin Province, No. 1745 Gongnong Road, Chaoyang District, Changchun 130021, China.
Yuchun
Y
Li
D000470:Alkaloids / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D020948:Cat's Claw / Q000737:chemistry*; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003377:Countercurrent Distribution / Q000379:methods*; D010936:Plant Extracts / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D014465:Ultrasonics / Q000379:methods*
D000470:Alkaloids; D010936:Plant Extracts; D012997:Solvents
Circulating ultrasound assisted extraction; Countercurrent chromatography; Extraction and online isolation; Uncaria tomentosa