Total Triterpenoid Extraction from Inonotus Obliquus Using Ionic Liquids and Separation of Potential Lactate Dehydrogenase Inhibitors via Ultrafiltration High-Speed Countercurrent Chromatography

Abstract

Extracts of the fungus Inonotus obliquus exhibit cytotoxic properties against different cancers; hence, this fungal species has been extensively studied. This study aimed to extract total triterpenoids from Inonotus obliquus using ionic liquids (ILs) and separate potential lactate dehydrogenase (LDH) inhibitors via ultrafiltration (UF)-high-speed countercurrent chromatography (HSCCC). Total triterpenoids from Inonotus obliquus were extracted by performing a single-factor experiment and employing a central composite design via ultrasonic-assisted extraction (UAE) and heat-assisted extraction (HAE). The extract was composed of 1-butyl-3-methylimidazolium bromide as the IL and methanol as the dispersant. Ultrafiltration-liquid chromatography (UF-LC) was used to rapidly scan the LDH inhibitors and betulin and lanosterol were identified as potential inhibitors. To obtain these target compounds, betulin and lanosterol with the purities of 95.9% and 97.8% were isolated from HSCCC within 120 min. Their structures were identified using several techniques, among which IL-HAE was fast and effective. This study reports the extraction of triterpenoids from Inonotus obliquus by IL for the first time. Collectively, the findings demonstrate that UF-LC is an effective tool for screening potential LDH inhibitors from crude extracts of I. obliquus and may help to identify bioactive substances against myocardial infarction, whereas high-purity compounds can be separated via UF-HSCCC.

Anti-Inflammatory Activity of Four Triterpenoids Isolated from Poriae Cutis

“…HSCCC is widely utilized in the isolating of natural products. It has high stability, good reproducibility, the non-requirement of a solid carrier, a low risk of nonreversible adsorption, and a high sample recovery rate [ 19 , 20 ]. Fractions of P. cocos were often obtained by means of a silica gel column chromatography and eluted successively [ 21 ], but this requires a complicated operation process and causes sample loss.…”

“…Fractions of P. cocos were often obtained by means of a silica gel column chromatography and eluted successively [ 21 ], but this requires a complicated operation process and causes sample loss. It was reported that HSCCC is an effective methodology for isolating of high-purity triterpenoid compounds in Inonotus obliquus [ 20 ]. In the present study, HSCCC was used to separate triterpenoid compounds from Poriae Cutis .…”

“…The 1 H- and 13 C-NMR spectra revealed H-11 (δ 5.21)/C-11 (δ 120.7), H-7 (δ 5.17)/C-7 (δ 119.0), C-9 (δ 138.3), C-8 (δ 142.3), H-18 (δ 0.99), H-19 (δ 0.93), H-26 (δ 1.50), H-27 (δ 1.58), H-29 (δ 1.68), H-30 (δ 1.50), H-16 (δ 4.54), H-28 (δ 4.67, 4.76), H-24 (δ 5.27), C-3 (δ 178.3), C-21 (δ 180.0). Based on the structural features of triterpene compounds and the data given in the literature, compound- 1 was identified as poricoic acid B [ 1 , 17 , 20 , 22 , 23 , 24 ]. Poricoic acid B belongs to the 3,4-seco-lanostan-7,9(11)-diene type triterpenes [ 4 ].…”

Design of experiment techniques for the optimization of chromatographic analysis conditions: A review

“…There is no doubt that more attention should be shown on the applications of DoE on these aspects. Some examples of DoE techniques applied for the development of analysis method in GC, TLC, and HSCCC are listed in Table 4 [111][112][113][114][115][116][117][118][119][120][121].…”

In Vitro Immunomodulatory Effects of Inonotus obliquus Extracts on Resting M0 Macrophages and LPS-Induced M1 Macrophages

“…Inonotus obliquus , also known as chaga, is a parasitic fungus that grows on birch trees and belongs to the Hymenochaetaceae family [ 1 ]. They are mainly distributed in northeast China and northern Russia [ 2 ]. Since the 16 th century, this fungus has been used as food and medicine for the prevention and treatment of malignant tumors, diabetes, and cardiovascular diseases in Russia, Poland, and the Baltic countries [ 3 ].…”