Abstract
Toddalia asiatica (L.) Lam. (Rutaceae) has shown a broad spectrum of biological properties, such as anti-inflammatory, antioxidant, antimicrobial, anti-HIV, and anticancer properties. The present study is concerned with the separation of the main components with broad partition coefficients (KD values) from T. asiatica, using linear gradient high-speed counter-current chromatography (LGCCC) combined with an off-line two-dimensional (2D) mode. Similar to the binary gradient HPLC, the LGCCC mode is operated by the adjustment of the proportion between the mobile phase of 5:5:1:9 (v/v) (pump A) and 5:5:4.5:5.5 (v/v) (pump B) in an n-hexane/ethyl acetate/methanol/water solvent system. The off-line 2D-CCC mode was used in this study for the secondary separation of two similar KD value compounds with n-hexane/ethyl acetate/methanol/water (5:5:4:6, v/v). Notably, six coumarins, namely, tomentin (1), toddalolactone (2), 5,7,8-trimethoxycoumarin (3), mexoticin (4), isopimpinellin (5), and toddanone (6), were efficiently separated. The structures of the pure compounds were elucidated by spectral techniques and compared with the literature.
Highlights
Toddalia asiatica (L.) Lam. (Rutaceae) is used in traditional medicine and is reported to treat wounds, pain, swelling, fever, malaria, diarrhea, cancer, coughs, and stomach infections
Previous studies show that different methods, such as GC/MS [3,13,14,15], HPLC [16,17,18], UPLC-MS/MS [19] UPLC-QTOF-MS/MS [17], and HPTLC [20], can be used for the purification of the chemical constitutes in T. asiatica
The current study demonstrates an alternate and more efficient separation method using two-dimensional HSCCC linear gradient and isocratic elution modes for the separation of the main components from T. asiatica
Summary
Toddalia asiatica (L.) Lam. (Rutaceae) is used in traditional medicine and is reported to treat wounds, pain, swelling, fever, malaria, diarrhea, cancer, coughs, and stomach infections. Previous studies show that different methods, such as GC/MS [3,13,14,15], HPLC [16,17,18], UPLC-MS/MS [19] UPLC-QTOF-MS/MS [17], and HPTLC [20], can be used for the purification of the chemical constitutes in T. asiatica. When the solvent system n-hexane/ethyl acetate/methanol/water (5:5:1:9, v/v) was used, the KD values of compounds 1 and 2 were suitable, whereas compounds 3–6 were larger than 5 and needed a long elution time. When the solvent system n-hexane/ethyl acetate/methanol/water (5:5:4:6, v/v) was used, the KD values of compounds 3 and 4 were suitable, whereas compounds 1 and 2 were smaller than 0.5 and had a poor resolution. When the solvent system n-hexane/ethyl acetate/methanol/water (5:5:5:5, v/v) was used, the KD values of compounds 5 and 6 were suitable, whereas compounds 1–4 were smaller. According to the results of the tested KD values, the lower phase of n-hexane/ethyl acetate/methanol/water (5:5:1:9, v/v) was used as mobile phase A, while n-hexane/ethyl acetate/methanol/water (5:5:5:5, 5:5:6:4, v/v) was selected as mobile phase B to build the 5:5:X:10-X (v/v) solvent system for LGCCC elution
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
