Efficient Separation of the Methoxyfuranocoumarins Peucedanin, 8-Methoxypeucedanin, and Bergapten by Centrifugal Partition Chromatography (CPC)

Abstract

Pure methoxyfuranocoumarins were isolated from a crude petroleum ether extract (CPE; Soxleth extraction efficiency 12.28%) from fruits of Peucedanum tauricum MB. (Apiaceae) by counter-current chromatography in a hydrostatic equilibrium system (centrifugal partition chromatography—CPC). The optimized biphasic solvent system composed of n-heptane-ethyl acetate-methanol-water (5:2:5:2; v/v/v/v) in the ascending mode of elution was used (3 mL/min, 1600 rpm). In the single run, peucedanin (P), 8-methoxypeucedanin (8MP), and bergapten (5MOP) were obtained as pure as 95.6%, 98.1%, and c.a. 100%, respectively. The carefully optimized and developed CPC was effectively transferred from the analytical to the semi-preparative scale (where 20 mg and 150 mg of CPE were loaded, respectively). Identification and quantitative analysis of methoxyfuranocoumarins was carried out in the plant material, in the CPE, and in individual CPC fractions by use of validated high-performance liquid chromatography with diode array detection and mass spectrometry (HPLC-DAD-ESI-MS). For the separation steps, the extraction/isolation recovery was calculated. In this case, CPC proved to be an effective tool for the simultaneous isolation and separation of P, 8MP, and 5MOP from a multicomponent plant matrix, without additional pre-purification steps. The high purity of the obtained plant metabolites makes it possible to consider their use in pharmacological or biological studies.

Evaluation of the Biological Effect of Non-UV-Activated Bergapten on Selected Human Tumor Cells and the Insight into the Molecular Mechanism of Its Action

“…99.0%) was isolated from CPE (crude petroleum ether extract) derived from P. tauricum fruits, and its identity and purity were confirmed via HPLC-DAD and LC-MS analyses (ESI-MS [H+] (data presented in the Supplementary File, Figures S1-S3). The fragmentation m/z was C12H8O4, MW 216.0901 ESI-MS: m/z 217.0974 (100, [M + H]+), 174.0436 (4.64), 146.0440 (2.52) 118.0484 (10.4), with the use of the standard compound and the literature's data [16], as described previously [17]. The isolated bergapten was used in the biological experiments described in this paper.…”

“…The isolation of 5-MOP from CPE was performed with the use of optimized centrifugal partition chromatography (CPC), as described previously [17]. Briefly, the CPC instrument employed in the present study was a model Armen SCPC-250-L (Armen Instrument, Saint Ave, France) equipped with an SCPC-250 Teflon column with a total capacity of 250 mL that was integrated with a gradient flow pump and a UV lamp (Flash 06S DAD 600) operating at various wavelengths.…”

Quantitative Analysis of Isopimpinellin from Ammi majus L. Fruits and Evaluation of Its Biological Effect on Selected Human Tumor Cells

“…Additionally, the easy transition from the analytical to the preparative scale is a big advantage of the CPC methodology [ 9 , 10 , 11 , 12 ]. CPC has been successfully used for the isolation of coumarins many times before, including in the work of our team [ 12 , 13 ].…”

“…This plant material serves as an excellent source of furanocoumarins, especially bergapten (5 MOP), xanthotoxin (8 MOP), and isopimpinellin (IsoP) [ 14 , 15 ], whose content was qualitatively and quantitatively evaluated in several studies. HPLC, LC/MS, and UPLC/MS methods were successfully developed for this purpose [ 13 , 18 , 19 ]. However, the PLE/ASE/HPLC methodology for quantitative or preparative purposes has not been used for A. majus fruit matrix to date, which was confirmed by the search in multiple databases (SciFinder CAS, Science Direct, Web of Science Clarivate, Medline, Pub-Med) (years 1990–2024; search topics; “(Ammi majus) and ASE”; ‘(Ammi majus) and PLE”).…”

GC-MS Analysis of Essential Oil and Volatiles from Aerial Parts of Peucedanum tauricum M.B. during the Phenological Period

“…which is a rare perennial plant, growing in nature on dry hillsides and in pinewoods in the Crimea, the Caucasus, and Romania [1,11,12]. Coumarins [13][14][15][16][17][18][19], phenolic acids [20], flavonoids [21] and essential oils (obtained by hydrodistillation of dried ripe fruit) [22] have been previously studied in this species, resulting in the isolation of two new compounds from the essential oil of its fruits [16,17]. Interesting changes of coumarin content during development of the plant were previously detected in aerial parts of P. tauricum [18].…”