• Korean Journal of Pharmacognosy
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• v.48 no.1
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• pp.93-96
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• 2017

Ephedra alkaloids, (-)-ephedrine, (+)-pseudoephedrine, (-)-N-methylephedrine, (+)-N-methylpseudoephedrine, (-)-norephedrine and (+)-norpseudoephedrine, from ephedra herb are sympathomimetic agonists causing an increase of metabolism, blood pressure and perspiration. In this study, we developed the validation method of (+)-pseudoephedrine and (-)-ephedrine, two major ephedra alkaloids in Ephedra spp., by high-performance liquid chromatography-ultraviolet spectrometer (HPLC-UV). HPLC analysis was performed using a HECTOR-M C18 column operating at $35^{\circ}C$, and UV detection at 215nm. The mobile phase used a gradient flow with 25 mM SDS in water (A) and acetonitrile (B).

• Natural Product Sciences
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• v.25 no.2
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• pp.111-114
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• 2019

The herbal formula, DF-02, consisting of Ephedra intermedia and Rheum palmatum are used for the treatment of the metabolic diseases such as obesity and liver fibrosis in Korean local clinics. We aimed to develop the simultaneous analytical conditions for four standards, (+)-pseudoephedrine (PSEP) and (-)-ephedrine (EP) for E. intermedia, and aloe-emodin (AE) and chrysophanol (CP) for R. palmatum using HPLC-UV techniques. The validated conditions yielded the high precision (relative standard deviation (RSD) < 3.65%) and the recoveries (94 - 106%) using the calibration curves with high linearity ($R^2$ > 0.9994). As a result, four standards of DF-02 were simultaneously determined under the developed method, which will be utilized for the quality control or evaluation of DF-02 and many herbal preparations containing E. intermedia and R. palmatum.

• Natural Product Sciences
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• v.28 no.4
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• pp.168-180
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• 2022

Ephedra is a genus of the Ephedraceae family and is found in temperate regions, such as Central Asia and Europe. Among the various ephedra species, Ma Huang (Ephedra herb) is derived from the aerial parts of Ephedra sinica S tapf, Ephedra equisetina Bunge, and Ephedra intermedia Schrenk & C.A. Mey. Ma Huang contains various ephedra alkaloids, including (-)-ephedrine, (+)-pseudoephedrine, (-)-norephedrine, (+)-norpseudoephedrine, (-)-methylephedrine, and (+)-methylpseudoephedrine, which are found naturally as single enantiomers, although they can be prepared as racemates. Although the use of Ma Huang in foods is prohibited in Korea, products containing Ma Huang can be imported, and so it is necessary to develop a suitable analytical technique for the detection of Ma Huang in foods. Herein, we report the development of analytical methods for the detection of ephedra alkaloids in products containing Ma Huang. Following sample purification by solid phase extraction, quantitative analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS). Additionally, the enantiomers were successfully separated using HPLC-DAD. We successfully analyzed various food samples, where the ephedra alkaloids were qualitatively and quantitatively determined, and the enantiomers were separated. It is expected that these methods may contribute toward preventing the distribution of illegal products containing Ma Huang.

• Natural Product Sciences
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• v.23 no.1
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• pp.9-15
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• 2017

DF formula is comprised of three traditional herbs, Ephedra intermedia, Rheum palmatum and Lithospermum erythrorhizon, and locally used for treating of the metabolic diseases, such as obesity and diabetes in Korea. We tried to optimize the extraction conditions of two major components, (-)-ephedrine and (+)-pseudoephedrine, in DF formula using response surface methodology with Box-Behnken design (BBD). The experimental conditions with 70% for EtOH concentrations, 4.8 hour for extraction hours and 8.7 times for the solvent to material ratio were suggested for the optimized extraction of DF formula with the highest amounts of (-)-ephedrine and (+)-pseudoephedrine in the designed model.

• Korean Journal of Pharmacognosy
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• v.53 no.4
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• pp.226-233
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• 2022

Ephedra herbs are defined as stem of Ephedra sinica , Ephedra intermedia and Ephedra equisetina in the Korean Pharmacopoeia. It is important to use pure herbs to derive the safety and efficacy of herbal medicine. However, the identification of these herbs by conventional taxonomic methods is difficult. Recently, many studies have applied these DNA barcoding for the identification of herbal medicinal species using standard DNA markers. In this study, we report a case study in which the identification of Ephedra species was done by DNA barcoding. For identification of Ephedra species, 17 samples were collected, and a reference DNA barcode library was developed using 6 markers (rbcL, matK, ITS2, ycf1, ycf3, and rpoC2). To develop KASP-SNP markers, we selected 4 markers (ycf1, ycf3, rpl2, and rbcL), which were able to distinguish three Ephedra species. In the result, the specific markers for each of the three Ephedra were clustered into FAM-positive section, whereas non-targeted plants were clustered either HEX-positive or negative section. Therefore, we have developed KASP assay that allow rapid and easy Ephedra species identification using three KASP markers.

• Journal of Korean Medicine for Obesity Research
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• v.23 no.1
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• pp.28-41
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• 2023

Objectives: This study was aimed to evaluate the hemodynamic feasibility using pulse parameters as a way to establish safe dose guidelines for herbal prescription containing Ephedra herb (Ephedra intermedia Schrenk & C.A.Mey) on weight loss, and to provide a foundation for developing evidence-based guidelines for clinical use. Methods: Forty-two volunteers were recruited to participate in a study examining the changes in pulse wave characteristics following the ingestion of Gambi-hwan, a herbal prescription containing ephedra, over a period of 4 weeks, and pulse wave measurements were taken before and after the administration. Pulse wave parameters were measured in this study using a 3-dimensional radial pulse tonometry device (DMP-Lifeplus). In addition, questionnaire, blood pressure, temperature, and body composition were also measured as secondary measures. Results: Fifteen minutes after administration of Gambi-hwan, the non-adverse event group (non-AE) exhibited a statistically significant increase in several power and pressure-related parameters, including h1, h5, systolic area, pulse area, and pulse width, while the AE group showed a trend of decreasing stroke volume and increasing Radial Augmentation Index (RAI), w, and w/t. After 4 weeks of administration, both groups exhibited significant changes in pulse rate, w/t, RAI, t3/t, stroke volume (SV), and stroke volume Index (SVI). Notably, there are significant differences between AE group and non-AE group in w/t, SV, and SVI. Conclusions: These findings suggest that pulse parameters may be a useful way to establish safe dosing guidelines for weight loss herbal prescription containing ephedra. Further research is needed to confirm these results and to develop evidence-based guidelines for clinical use.

• Biomedical Science Letters
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• v.23 no.2
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• pp.144-153
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• 2017

It was demonstrated that Gangjihwan (DF), which is the herbal composition composed of Ephedra intermedia, Lithospermum erythrorhizon, and Rheum palmatum, inhibits obesity and hepatic steatosis in high fat diet (HFD)-fed obese mice. The aim of this study was to determine the effects of DF on visceral obesity, hepatic inflammation and fibrosis and the mechanism of actions involved in this process using in vivo and in vitro approaches. DF was extracted with water (DF-FW), 30% grain alcohol (DF-GA30), and 70% grain alcohol (DF-GA70). Administration of DF to HFD-fed control mice decreased visceral tissue mass and visceral adipocyte size without adverse effects. Visceral fat mass was decreased by DF-GA30 and DF-GA70, and visceral adipocyte size by all three DF extracts compared with obese control mice. Histological analysis revealed that three kinds of DF extracts reduced toluidine blue-stained mast cells and collagen accumulation in the liver, the extents of which were most eminent in DF-GA70-treated mice. DF-GA70 decreased the mRNA levels of the inflammation ($TNF{\alpha}$ and VCAM-1), fibrosis (${\alpha}-SMA$), and apoptosis (caspase 3) genes, but increasing the anti-apoptosis gene (Bcl-2) mRNA levels in the liver of obese control mice. Consistent with the in vivo data, GA-70 also altered the expression of inflammation genes ($TNF{\alpha}$ and MCP-1) in HepG2 cells. These results indicate that DF not only inhibits visceral obesity, but also ameliorates visceral obesity-induced hepatic inflammation and fibrosis and that this process may be mediated by regulating the hepatic expression of inflammatory and fibrogenic genes.

• The Korea Journal of Herbology
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• v.34 no.6
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• pp.25-32
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• 2019

Objectives : Ephedra regeliana, although similar to the other E. sinica, E. intermedia, and E. equisetina, is not authorized by The Korean Pharmacopoeia and yet has also been imported under the name of 'Ma-Hwang'. The aim of this study was to perform a comparative anatomical evaluation of Ephedrae Sinicae Herba (ESH), Ephedrae Intermedicae Herba (EIH), Ephedrae Equisetinae Herba (EEH) and Ephedrae Regelianae Herba (ERH). Methods : Permanent paraffin-embedded specimens were produced using the paraffin-tertiary butyl alcohol (TBA) method, to visualize their anatomical features via light microscopy. Results : ESH and ERH transverse sections were in the shape of a long ellipse, while those of EIH and EEH were in the shape of a circle. These anatomical characteristics substantiated the results of external morphological examination. The sections revealed that each of the four samples showed significantly different cortexes. The cortex of ESH was wider than two thirds of the section, unlike that of the other samples which was relatively narrow. The xylems of ERH, EIH, and EEH demonstrated interconnectivity with other adjacent xylems, contrary to those in ESH. Therefore, the characteristic development of the xylems over time was used to distinguish ERH from the other. Furthermore, parenchymal morphology in the central cylinder was also a good criterion to differentiate between EIH and EEH, since EIH demonstrated parenchymal cells that were shaped like a rounded-triangle, whereas EEH parenchymal cells were chrysanthemum-shaped. Conclusions : The identification-keys established in this study would facilitate the accurate identification of microscopic features of the four varieties of 'Ma-Hwang'.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.4
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• pp.1116-1119
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• 2003

Ma huang, the dried plant stem of Ephedra Intermedia Schrenk et CA, is one of the well known medicinal herbs, and has been used for the diaphoretic, antiasthmatic, and diuretic actions. Ma huang is an ephedrine type alkaloid used for the weight loss and energy expenditure. Medications based on the Ma huang have been found to be effective in the treatment of obesity. Neuropeptide Y (NPY), a 36-amino-acid peptide and concentrated in the hypothalamus, stimulates feeding desire and decrease energy expenditure. In the present study, the effect of Ma huang on the expression of NPY in the rat hypothalamus was investigated using immunohistochemistry. The present results demonstrated that Ma huang treatment suppressed weight gain and NPY expression in the hypothalamus depending upon the dosage used. Based on the results, it can be suggested that Ma huang treatment is effective in curbing the desire for food via modulation of NPY expression under the normal conditions.