• Korean Journal of Pharmacognosy
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• v.33 no.2 s.129
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• pp.96-99
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• 2002

20-Hydroxyecdysone was isolated from Melandrii Herba and identified by the spectroscopic methods. In order to evaluate the quality of it, quantitative determination of 20-hydroxyecdysone in Melandrii Herba using HPLC method has been conducted. Content of 20-hydroxyecdysone in Melandrii Herba showed average 0.0138% in 42 samples collected throughout the regions of Korea.

• The Korea Journal of Herbology
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• v.23 no.1
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• pp.109-116
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• 2008

Objectives : This study was performed to investigate the discriminative criteria of 6 kinds of Achyranthis Radix by HPLC/DAD. Methods : 20-hydroxyecdysone is isolated by silica gel column chromatography ($CHCl_3$:MeOH, 7:1-1:1 v/v) and identified by nuclear magnetic resonance, A high-performance liquid chromatographic method with diode array detection was used to identify 20-hydroxyecdysone in A. japonica. The analysis was performed using $C_{18}$ column with isocratic elution consisted of 18% acetonitrile and 82% water and the detection was carried out by DAD at 254 nm. 6 kinds of Achyranthis Radix from different locations were extracted in MeOH. Each extracts was analyzed by HPLC in same condition as used in analysis of 20-hydroxyecdysone. The identities of each extracts were determined by comparing the retention time and UV spectrum with that of reference compound. Results : 1. A. japonica and A. bidentata showed the similar patterns of HPLC chromatogram and 20-hydroxycedysone was present in both of them because the peaks having the same retention time and UV spectrum as 20-hydroxyecdysone were shown in the HPLC chromatograms of A. japonica and A. bidentata 2. Cyathula officinalis and C. capitata showed the similar patterns of HPLC chromatogram. The peak having the same retention time and UV spectrum as 20-hydroxyecdysone was shown in the HPLC chromatogram of C. capitata but not shown in the HPLC chromatogram of C. officinalis. 3. Two species of medicinal drugs from Sacheon province showed similar patterns of HPLC chromatogram. Achyranthis Radix from Sacheon(wild) did not have 20-hydroxycedysone but Achyranthis Radix from Sacheon(cultivated) showed the peak having the same retention time as 20-hydroxyecdysone but UV spectrum of the peak was different from that of 20-hydroxyecdysone. Conclusions : These results suggested that 20-hydroxyecdysone could be the discriminative criteria for Achyranthis Radix contain 20-hydroxyecdysone though they belong to different genus and species. And the patterns of HPLC chromatogram also could be the discriminative criteria as the different species of Achyranthis Radix belonging to the same genus showed similar patterns of HPLC chromatogram.

• Korean Journal of Pharmacognosy
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• v.30 no.3
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• pp.335-339
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• 1999

Separation and quantitative determination of 20-hydroxyecdysone from Achyranthis Radix has been conducted by using HPLC method. 20-hydroxyecdysone in a methanol extract from the raw drug was separated on a reverse phase column using a $CH_3CN-H_2O\;(18:82)$ solvent system and the average content is $0.0931{\pm}0.0048%$. For the preparation of authentic standard, we isolated 20-hydroxyecdysone from the roots of Achyranthes fauriei by $SiO_2$ and RP-18 column chromatography.

• Korean journal of applied entomology
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• v.34 no.4
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• pp.287-294
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• 1995

Studies were carried out to elucidate the efficacy of exogeneous 20-hydroxyecdysone treatment on terminating pupal diapause in the fall webworm. Hyphantria cunea Drury. And the difference was also investigated between normal adult development and post-diapause development after 20-hydroxyecdysone treatment. In the diapause termination rate of pupae treated with 20-hydroxyecdysone after storage for various periods at 16L:8D and $25\pm$$1^{\circ}C$, at the highest rate was observed from the group stored for the longest period and the lowest rate from those stored for 1.5 months. The time needed for adult emergence was inversely proportional to the chilling (at $0\pm$$1^{\circ}C$) period, and the longer its exposure period at low temperature, the higher its sensitivity to 20-hydroxyecdysone treatment. Pupal diapause duration was almost the same, regardless of storage period in the total darkness or at the photoperiod of 16L:8D, and also they successfully emerged to adult even without any experience at low temperature. The oxygen consumption rate in normally developing pupae showed nearly a typical U-form. But, that of diapausing pupae treated with 20-hydroxyecdysone slowly increased and jumped 14 days after the treatment. Pupal diapause began before formation of adult tissues, an a timing of adult tissue formation coincided with ascending timing of the metabolic rate in both normally developing pupae and diapausing pupae treated with 20-hydroxyecdysone. The diapausing pupae treated with 20-hydroxyecdysone was similar to normally developing pupae in band patterns of proteins from haemolymph or fat body.

• The Korean Journal of Zoology
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• v.36 no.3
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• pp.373-379
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• 1993

The heat shock protein (HSP) genes are expressed at various stages of the Drosophila life cycle even under non-heat-shock conditions. In the present study, developmental changes of HSP23 gene expression and the role of 20-hydroxyecdysone (2OHE) on the HSP23 gene expression were investigated in Drosophila melanogaster. The Northern blot and Western blot analyses showed that HSP23 gene expression occurred at the early third instar larval stage, reached the highest with a sharp peak in the white prepupa, and then decreased throughout the pupal period. When the HSP23 gene expression was compared with the secretion of 20HE, there is a slmiladty between 20HE synthesis and HSP23 gene expression during the third instar larval-prepupal period. It appears that 2OHE regulates expression of HSP23 gene at larva-pupa molting period, and that, 2OHE is also involved in the control the metamorphosis in some part through the HSP23.

• Korean Journal of Pharmacognosy
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• v.39 no.4
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• pp.316-323
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• 2008

In this study, we have investigated the HPLC analysis methods and quantitative analysis of standard compounds for quality standardization of a medicinal crude drug GCSB-5, a herbal formulation consisting of 6 medicinal plants (Acanthopanacis Cortex, Achyranthis Radix, Ledebouriellae Radix, Cibotii Rhizoma, Glycine Semen, Eucommiae Cortex) which are used in traditional medicine to treat various bone disorders. HPLC analysis methods of acanthoside D(Acanthopanacis Cortex), 20-hydroxyecdysone(Achyranthis Radix) which were known standard compounds among 6 medicinal plants were developed on crude material and product. And validation of HPLC analysis methods were conformed for verification of HPLC methods by check to specificity, linearity, intra-day precision, inter-day precision and accuracy following ICH guideline. Content of acanthoside D and 20-hydroxyecdysone on raw material of GCSB-5 were decided at 0.577-0.578 mg/g and 0.311-0.312 mg/g. And we confirmed that content of acanthoside D and 20-hydroxyecdysone on GCSB-5 preparation were 0.302-0.303 mg/capsule and 0.113-0.115 mg/capsule.

• 한국약용작물학회:학술대회논문집
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• 2007.05a
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• pp.263-264
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• 2007

• Korean Journal of Pharmacognosy
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• v.29 no.4
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• pp.318-322
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• 1998

Continuing to previous report, seven compounds were isolated from the aerial parts of Ajuga multiflora. The structures of them were established as Di-2-ethylhexyl phthalate (1), ursolic acid (2), sterol glucoside (3), 20-hydroxyecdysone (4), makisterone A (5). cyasterone (6) and apigenin 7-glucuronide (7), respectively.

• Korean journal of applied entomology
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• v.27 no.1
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• pp.6-13
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• 1988

Studies were carried out to investigate the diapause-sensitive stage, the effect of 20-hydroxye- cdysone on diapausing pupae and the changes in pupal proteins of the fitll webworm, HYPha¬ntria cunea Drury. The stage sensitive to diapause-inducing photoperiod lies between 5th and 15th day of the larval development. And the oxygen consumption rate of diapausebound pupae decreased to a mean level of $30mu$l $O_{2}$/g/hr. The 20-hydroxyecdysone was effective in raising the respiratory acivity of 2.5% homogenate of the diapausing fall webworm pupae and in causing a normal adult development of the diapause-bound pupae. The soluble protein content rapidly declined in normally-developing pupae, but in diapause-bound pupae it was more or less static with much higher quantity. And protein bands separated from fat body of diapause¬bound pupae were different from those of nondiapausing pupae.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.2
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• pp.67-75
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• 2020

Objectives SJ004 is a natural herbal medicine that contains Acyranthes japonica Nakai and Eucommia ulmoides Oliver traditionally used for joint and spinal diseases. This study aimed to establish an efficient method of extracting SJ004 to standardize using the yield, high-performance liquid chromatography (HPLC), and antioxidant assay. Methods SJ004 was extracted with distilled water, 70% and 100% of ethyl alcohol (EtOH). The method validation of 20-hydroxyecdysone and pinoresinol diglucoside was determined by HPLC-photo diode array and the content of SJ004 was calculated. The antioxidant activity of each extract was compared and measured using total flavonoids, total phenolic compounds, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power according to the standard protocol. Results The yield was highest in pure water extract and lowest in 100% EtOH. But, the content of marker compounds indicating 20-hydroxyecdysone and pinoresinol diglucoside was highest in 100% EtOH extract. In the physiological activity measurement using antioxidant activity, 100% ethanol extract was highest. The limit of detection indicating 20-hydroxyecdysone and pinoresinol diglucoside were analyzed 0.33 ㎍/mL, 0.1616 ㎍/mL, and the limit of quantification were analyzed 1.01 ㎍/mL and 0.49 ㎍/mL respectively. Conclusions The experimental results showed that the extraction conditions have a significant effect on content of marker compounds and antioxidant activity. As a result of method validation, SJ004 was standardized by 20-hydroxyecdysone and pinoresinol diglucoside.