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Cervi parvum cornu (CPC) is a well-known ethnopharmacological source, whereas Rangifer cornu (RC) is not considered to be a major source. CPC is distributed in sliced form. Addition of RC to CPC has become an issue in CPC distribution because the appearance of sliced RC is not different from sliced CPC. Therefore, a real-time polymerase chain reaction (PCR) method was developed in this study to detect contaminating RC in CPC. The C-VIC and R-FAM primer/probe sets were designed to specifically amplify CPC and RC DNA, respectively. The specificities and sensitivities of real-time PCR using two primer/probe sets and the applicability of the real-time PCR to powder mixtures, which involved mixtures of powdered CPC and powdered RC in diverse ratios, were evaluated. Real-time PCR using C-VIC and R-FAM primer/probe sets specifically and sensitively amplified both CPC and RC DNA. Furthermore, real-time RCR sensitively detected RC DNA in the powder mixtures of CPC and RC. These results indicate that this real-time PCR method using two primer/probe sets is sufficiently applicable for the detection of contaminant RC in CPC.

Although it has been known that pomegranate (Punica granatum L.) contains several steroid hormones, concrete experimental proofs about that have not been published until now. In order to identify and evaluate the contents of steroid hormones including estrone in pomegranate, we analyzed pomegranate seed, fruit juice and commercial preparations. We developed a reproducible and sensitive method for separation and identification of steroid hormones in pomegranate samples using both high performance liquid chromatography (HPLC)–PDA and gas chromatography (GC)–MS. In case of HPLC, an isocratic elution method using 35% aqueous acetonitrile solution at 1.0 ml/min with photodiode-array (PDA) detection at 225 nm and 254 nm was found to optimally separate and identify the steroid hormones from the pomegranate samples with a run time of less than 30 min. The pomegranate samples were comparatively analyzed to the HPLC results by GC/FID or GC/MS detection on a HP-1 (30 m length, 0.32 mm I.D.) with helium as carrier gas under the oven temperature control as follows: start 220 °C for 5 min, raising 5 °C per min, final 280 °C for 10 min. The HPLC and GC methods were successfully applied to the identification of steroid hormones in pomegranate samples. Our results suggested that there were no steroid estrogens including estrone, estradiol and testosterone in pomegranate seed, fruit juice and preparations. Consequently, we assumed that the previously reported analysts of pomegranate were misunderstood their analytical results according to either the estrogen-like effects or similarity of peak retention time and Rf values in experiments.

Three new ( 1–3 ) and two known ( 4–5 ) triterpene glycosides were isolated from the leaves of Acanthopanax japonicus (Araliaceae) and elucidated structurally by mass, 1D, and 2D NMR spectroscopy. All the compounds possessed a nor -oleanene triterpene skeleton as the aglycone. The structures of 1–5 were established as 28- O -α- l - rhamno-pyranosyl-(1→4)-β- d -glucopyranosyl-(1→6)-β- d -glucopyranosyl ester of 3β-hydroxy- 30- nor -olean-12,20(29)-diene-23,28-dioic acid, designated as acanjaposide A, 3β- hydroxy-23-oxo-30- nor -olean-12,20(29)-diene-28-oic acid, named acanjaposide B, 3β,20α-dihydroxy-23-oxo-30- nor -olean-12-en-28-oic acid, named acanjaposide C, and nipponoside E, a known saponin, respectively.

The metabolic pathway of chiisanoside isolated from leaves of Acanthopanax divaricatus var. albeofructus (Araliaceae) by human intestinal bacteria and by the protein fraction of leaves of this plant were investigated, and the cytotoxic and anti-rotaviral activities of chiisanoside and its metabolite, chiisanogenin, were assayed. Chiisanogenin was produced as a main metabolite, when chiisanoside were incubated for 15 h with human intestinal bacteria. This metabolic pathway proceeded more potently with the protein fraction than with human intestinal bacteria. The in vitro cytotoxicity of chiisanogenin was superior to that of chiisanoside. H+/K+ ATPase was more potently inhibited by chiisanogenin than by chiisanoside. However, the anti-rotaviral activity of chiisanoside was more potent than that of chiisanogenin.

Four new 3,4-seco-lupane-type triterpene glycosides (1-4) were isolated from the leaves of Acanthopanax senticosus forma inermis. The structures of 1-4 were established as 11alpha-hydroxy-3, 4-seco-lup-4(23),20(30)-dien-3-oic acid methyl ester 28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1--4)-beta-D-glucopyranosyl-(1--6)-be ta-D-glucopyranoside, designated as inermoside (1); 1-deoxychiisanoside (2); 24-hydroxychiisanoside (3); and 11-deoxyisochiisanoside (4) by (1)H-(1)H COSY and (1)H-(13)C COSY(HMBC, HMQC) methods and FABMS.

Sulfite was detected in 7 varieties of Oriental herbal medicines (Pueraria radix, Zingiberis rhizoma, Platycodon radix, Adenophora radix, Pinellia tuber, Astragalus radix, and Paeonia radix) on the Korean market. Sulfiting of commercial Oriental herbal medicines by fumigation with burning bituminous coal was simulated, and the accumulation of sulfite was investigated by using fresh Platycodon radix roots obtained from a growing field. The sulfite level reached a plateau in 9 h, and the maximum sulfite level found by the Monier-Williams (MW) method (AOAC 990.28) was 1020 ppm. The sulfite content in the simulated Platycodon radix sample determined by alkali extraction followed by ion-exclusion chromatography with electrochemical detection (AOAC 990.31) was approximately 17% lower on average than the MW results. Free-sulfite levels determined by acid extraction and ion-exclusion chromatography with electrochemical detection were between 19 and 49% of the MW results. The advantages of different methods for sulfite determination and the significance of the results are discussed.

Two new lupane-triterpene glycosides named acankoreosides C and D, were isolated from the leaves of Acanthopanax koreanum. Based on spectroscopic data, the chemical structures were determined as 3-O-β- d -glucopyranosyl 3α,11α-dihydroxylup-20(29)-en-28-oic acid 28-O-α- l -rhamnopyranosyl-(1→4)-β- d -glucopyranosyl-(1→6)-β- d -glucopyranosyl ester and 3α,11α-dihydroxylup-23-al-20(29)-en-28-oic acid 28-O-α- l -rhamnopyranosyl-(1→4)-β- d -glucopyranosyl-(1→6)-β- d -glucopyranosyl ester, respectively.

A new lupane triterpene glycoside, acantrifoside A, was isolated from the leaves of Acanthopanax trifoliatus and A. koreanum . Based on spectroscopic data, the compound was identified as 3α,11α-dihydroxylup-20(29)-en-28-oic acid 28- O -α- l -rhamnopyranosyl-(1→4)-β- d -glucopyranosyl-(1→6)-β- d -glucopyranosyl ester.

Two new lupane-triterpene glycosides, acankoreoside A (I) and B (2), were isolated from the leaves of Acanthopanax koreanum NAKAI (Araliaceae). Based on spectroscopic data, the chemical structures of 1 and 2 were determined as 3α-hydroxy-lup-20(29)-en-23,28-dioic acid 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester and 3α,11α,23-trihydroxy-lup-20(29)-en-28-oic acid 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester, respectively.

Two androstane alkaloids were isolated from the musk of Moschus moschiferus. The structures were elucidated to be 3alpha-ureido-androst-4-en-17-one (1) and 3alpha-ureido-androst-4-en-17beta-ol (2) by two-dimensional NMR analysis (HMQC, 1H-1H COSY, HMBC, and NOESY).

Two 3,4-seco-lupane triterpenoids (1, 2) were isolated from the leaves of Acanthopanax divaricatus var. albeofructus (Araliaceae). Based on the spectroscopic data, the chemical structures of 1 and 2 were characterized as 24-hydroxychiisanogenin and 22alpha-hydroxychiisanogenin, respectively. 1 was a new compound and 2 was isolated for the first time from the natural source, although it had been obtained as an enzymatically hydrolyzed artifact from 22alpha-hydroxychiisanoside.

Two new lupane-triterpene glycosides, acankoreoside A (I) and B (2), were isolated from the leaves of Acanthopanax koreanum NAKAI (Araliaceae). Based on spectroscopic data, the chemical structures of 1 and 2 were determined as 3α-hydroxy-lup-20(29)-en-23,28-dioic acid 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester and 3α,11α,23-trihydroxy-lup-20(29)-en-28-oic acid 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester, respectively.

A new and two known lupane-triterpene glycosides were isolated from the hot MeOH fraction of the leaves of Acanthopanax gracilistylus W. W. Smith. Based on the physical properties and spectroscopic data, their chemical structures were determined as acankoreoside A (1), acankoreoside D (2), and 3alpha-hydroxy-lup-23-al-20(29)-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (3), respectively. To our best knowledge, compound 3 appears to be novel, which was named as wujiapioside A.

A novel lupane-triterpene glycoside, called wujiapioside B (1), was isolated from the leaves of Acanthopanax gracilistylus (Araliaceae) together with three known lupane-triterpene glycosides, acankoreoside C (2), acantrifoside A (3) and 3-epibetulinic acid 28-O-alpha-L-rhamnopyranosyl-(1--4)-beta-D-glucopyranosyl-(1--6)-beta-D-glucopyranosyl ester (4). Based on spectroscopic data, the chemical structure of 1 was determined as 3alpha,23-dihydroxy-lup-20(29)-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1--4)-beta-D-glucopyranosyl-(1--6)-beta-D-glucopyranosyl ester. Compounds 2-3 were obtained for the first time from this plant and compound 4 has not been isolated from Acanthopanax genus yet.

We report the development of enzyme-linked immunosorbent assay (ELISA) for the quantitative analysis of a unique musk protein (MP-1) in musk samples. Musk defatted with ethyl acetate/methanol (9:1, v/v) was dipped in cold water and ammonium sulfate was added to the supernatant up to 85% saturation. The resulting precipitate was applied to a Bio-Gel P-100 chromatography. The fraction eluted at the void region was collected and it was consecutively purified by affinity chromatography on a DEAE Affi-Gel Blue and on anion-exchange columns containing DEAE-Sepharose CL-6B. This protein was determined to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under denaturing conditions with an apparent molecular weight of 35000 Da and was called as musk protein-1 (MP-1). Polyclonal antibodies of MP-1 were produced by injecting it into a rabbit. These antibodies were reactive to the aqueous extract of musk and the pure antigen. The ELISA could be applied to detect nano gram quantities of the antigen in musk samples. This method made it possible to distinguish musk samples from different origins.

Two 3,4-seco-lupane triterpenoids (1, 2) were isolated from the leaves of Acanthopanax divaricatus var. albeofructus (Araliaceae). Based on the spectroscopic data, the chemical structures of 1 and 2 were characterized as 24-hydroxychiisanogenin and 22alpha-hydroxychiisanogenin, respectively. 1 was a new compound and 2 was isolated for the first time from the natural source, although it had been obtained as an enzymatically hydrolyzed artifact from 22alpha-hydroxychiisanoside.

Two androstane alkaloids were isolated from the musk of Moschus moschiferus. The structures were elucidated to be 3alpha-ureido-androst-4-en-17-one (1) and 3alpha-ureido-androst-4-en-17beta-ol (2) by two-dimensional NMR analysis (HMQC, 1H-1H COSY, HMBC, and NOESY).