Development of an efficient fractionation method for the preparative separation of sesquiterpenoids from Tussilago farfara by counter-current chromatography

Song, Keun Bae; Lee, Kyung Jin; Kim, Yeong Shik

doi:10.1016/j.chroma.2017.02.005

Cited by 22 publications

(16 citation statements)

References 34 publications

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“…ECN was isolated from dried buds of Tussilago farfara and identified, as previously reported by our group [13] . Fetal bovine serum (FBS), penicillin, and streptomycin were purchased from GenDepot (Barker, TX, USA).…”

Section: Methodsmentioning

confidence: 71%

Neuroprotection against 6-OHDA toxicity in PC12 cells and mice through the Nrf2 pathway by a sesquiterpenoid from Tussilago farfara

Lee

Song

Huh³

et al. 2018

Redox Biology

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Oxidative stress plays a key role in neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Therefore, the nuclear factor-E2-related factor 2 (Nrf2), a key regulator of the antioxidative response, is considered to be important as a therapeutic target for neurodegenerative diseases. We investigated the underlying mechanism of Nrf2-mediated neuroprotective effects against oxidative stress in the PC12 cell line by 7β-(3-ethyl-cis-crotonoyloxy)-1α-(2-methylbutyryloxy)-3,14-dehydro-Z-notonipetranone (ECN), one of the sesquiterpenoids in Farfarae Flos. Pretreatment of PC12 cells with ECN had a protective effect against hydrogen peroxide (H2O2)- or 6-hydroxydopamine (6-OHDA)-induced cytotoxicity. ECN upregulated the ARE-luciferase activity and induced the mRNA expression of Nrf2 and antioxidant enzyme heme oxygenase-1 (HO-1). Knockdown of Nrf2 by small, interfering RNA (siRNA) abrogated the upregulation of HO-1, indicating that ECN had induced HO-1 via the Nrf2 pathway. Pretreatment with the thiol reducing agents, N-acetylcysteine (NAC) or dithiothreitol (DTT), attenuated Nrf2 activation and HO-1 expression. However, the non-thiol reducing antioxidant, Trolox, failed to inhibit HO-1 induction by ECN. These results suggest that ECN may directly interact with Kelch-like ECH-associated protein 1 (Keap1) and modify critical cysteine thiols present in the proteins responsible for Nrf2-mediated upregulation of HO-1. In a 6-OHDA-induced mouse model of PD, administration of ECN ameliorated motor impairments and dopaminergic neuronal damage. Taken together, ECN exerts neuroprotective effects by activating the Nrf2/HO-1 signaling pathway in both PC12 cells and mice. Thus, ECN, as an Nrf2 activator, could be an attractive therapeutic candidate for the neuroprotection or treatment of neurodegenerative diseases.

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Section: Methodsmentioning

confidence: 71%

Neuroprotection against 6-OHDA toxicity in PC12 cells and mice through the Nrf2 pathway by a sesquiterpenoid from Tussilago farfara

Lee

Song

Huh³

et al. 2018

Redox Biology

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“…ECN (purity ≥ 97.0%) was isolated according to our previous report [21]. Losartan (LST) potassium (IS; purity ≥ 99.5%) and ammonium formate (purity ≥ 99.0%) were purchased from Sigma-Aldrich (St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

“…Owing to its potent pharmacological activities, a method for the extraction and isolation of ECN from T. farfara has been investigated [21]. However, pre-clinical studies investigating ECN's pharmacokinetic properties have not been reported yet, and suitable analytical methods for its determination in biological fluids are not available.…”

Section: Introductionmentioning

confidence: 99%

Development and Validation of Liquid Chromatography-Tandem Mass Spectrometry Method for Pharmacokinetic Evaluation of 7β-(3-Ethyl-cis-crotonoyloxy)-1α-(2-methylbutyryloxy)-3,14-dehydro-Z-notonipetranon in Rats

et al. 2020

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Recently, potent neuroprotective and anti-diabetic effects of 7β-(3-Ethyl-cis-crotonoyloxy)-1α-(2-methylbutyryloxy)-3,14-dehydro-Z-notonipetranone (ECN), a sesquiterpenoid isolated from Tussilago farfara Linnaeus, have been elucidated. To facilitate further pre-clinical evaluation in rats, an analytical method for the determination of ECN in rat plasma was developed and optimized by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Plasma samples were pretreated by the protein precipitation method with an acetonitrile solution of losartan (LST) as the internal standard. Chromatographic separation was performed using a an Octadecyl-silica (ODS) column (2.6 µm, 100 x 4.6 mm) in the isocratic mode. The mobile phase, comprising 10 mM ammonium formate in water pH 5.75) and acetonitrile (11:89, v/v), was eluted at a flow rate of 0.4 mL/min. Mass spectrometric detection was performed in the multiple reaction monitoring mode with positive electrospray ionization, and the mass transitions of ECN and LST were m/z 431.3 to 97.3 and m/z 423.1 to 207.2, respectively. The calibration curves of spiked plasma samples were linear in the 10.0–10,000 ng/mL range (r2 > 0.996). The lower limit of quantification (LLOQ) was determined as 10.0 ng/mL. Validation was conducted in the LLOQ, and three quality control (QC) sample levels (10.0, 25.0, 3750, and 7500 ng/mL) were studied. Among them, the relative standard deviation for the within- and between-run precisions was under 9.90%, and the relative error of the accuracies was within the −8.13% to 0.42% range. The validated method was successfully employed to investigate the pharmacokinetic properties of ECN in rats, which revealed the linear pharmacokinetic behavior of ECN for the first time.

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“…Countercurrent chromatography (CCC) is a type of liquid chromatography that is used in the field of natural products to separate bioactive components. However, the separation and purification of compounds using conventional methods, such as adsorption chromatography, results in low recovery, low sample capacity, and poor resolution, the use of CCC produces more cost‐effective separations and high yields of the target compounds . These advantages are closely related to the fact that both stationary and mobile phases are liquids.…”

Section: Introductionmentioning

confidence: 99%

“…These advantages are closely related to the fact that both stationary and mobile phases are liquids. The principle of countercurrent separation is the partitioning of target compounds between two immiscible liquid phases according to their partition coefficients ( K D ) .…”

Section: Introductionmentioning

confidence: 99%

Application of linear gradient elution in countercurrent chromatography for the separation of triterpenoid saponins from the roots of Pulsatilla koreana Nakai

Lee

Song

et al. 2017

J of Separation Science

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A linear gradient elution method using countercurrent chromatography was developed for the separation of four triterpenoid saponins from the roots of Pulsatilla koreana Nakai, including hederacolchiside E, which is responsible for the neuroprotective activity of this plant. The target fraction was obtained by 80% methanol elution of solid phase column chromatography. The partition coefficients of the target compounds were very different, which means they are difficult to separate with a single biphasic solvent system. Several important parameters for gradient elution, such as addition of alcohol content to the solvent system, starting point of the second mobile phase, and the time for the gradient change were logically determined and optimized. Four triterpenoid saponins could ultimately be separated, analyzed by high-performance liquid chromatography, and their structures were identified by comparing the mass spectra and NMR spectra with the literature data. The compounds and yields were: hederasaponin B (1; 21.3 mg/100 mg), hederacolchiside E (2; 19.8 mg/100 mg), cernuoside A (3; 18.4 mg/100 mg), and cernuoside B (4; 17.3 mg/100 mg). Gradient-elution countercurrent chromatography allows the effective separation of compounds with a wide polarity range.

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Development of an efficient fractionation method for the preparative separation of sesquiterpenoids from Tussilago farfara by counter-current chromatography

Cited by 22 publications

References 34 publications

Neuroprotection against 6-OHDA toxicity in PC12 cells and mice through the Nrf2 pathway by a sesquiterpenoid from Tussilago farfara

Neuroprotection against 6-OHDA toxicity in PC12 cells and mice through the Nrf2 pathway by a sesquiterpenoid from Tussilago farfara

Development and Validation of Liquid Chromatography-Tandem Mass Spectrometry Method for Pharmacokinetic Evaluation of 7β-(3-Ethyl-cis-crotonoyloxy)-1α-(2-methylbutyryloxy)-3,14-dehydro-Z-notonipetranon in Rats

Application of linear gradient elution in countercurrent chromatography for the separation of triterpenoid saponins from the roots of Pulsatilla koreana Nakai

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