Young Pyo Jang

Direct analysis of curcumin in turmeric by DART‐MS

INTRODUCTION The new ion source technique, direct analysis in real time (DART) atmospheric pressure ionisation, allows high resolution mass measurements of gas, liquid and solid samples. As DART-MS produces [M + H](+) molecular ions of most compounds, relatively simple and clear mass spectra are obtained even of multi-component samples. In order to take advantage of the capacity of DART-MS for the real time analysis of individual compounds in natural raw materials, a pilot study was performed using the well-known antioxidant botanical drug, turmeric. OBJECTIVE To establish the analysis methods of curcumin and its derivatives from various types of samples with DART-MS and compare the efficiency of the method with traditional HPLC method. RESULTS Different curcuminoids were successfully detected directly from the raw particles of Curcuma longa. When a turmeric extract was separated on a TLC plate, each band produced molecular ion peaks corresponding to curcumin, demethoxycurcumin and bisdemethoxycurcumin. Molecular ions of curcuminoids in turmeric-containing beverages and curry powder were also efficiently detected. In addition to high efficiency of qualitative analysis, the evaluation of its linearity showed that DART-MS can be applied for semi-quantitative determinations of curcumin over a large range (5-100 microg/mL). CONCLUSION A simple chemical profiling and semi-quantitative method for natural products using DART-MS might be applied to diverse field related quality control of medicinal plants or food ingredients.

Inhibition effect of Gynura procumbens extract on UV-B-induced matrix-metalloproteinase expression in human dermal fibroblasts

ETHNOPHARMACOLOGICAL RELEVANCE Gynura procumbens Merr. (Asteraceae) has been used as a traditional remedy for various skin diseases in certain areas of Southeast Asia. AIM OF THE STUDY In order to evaluate the protective activity of Gynura procumbens extract on skin photoaging and elucidate its mode of action. MATERIALS AND METHODS Matrix-metalloproteinase (MMP)-1 and -9 expressions were induced by UV-B irradiation in human primary dermal fibroblasts. MMP-1 expression level was measured by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Zymography was employed for evaluating the enzymatic activity of MMP-9. Anti-inflammatory activity and anti-oxidative capacity of the extract were evaluated by ELISA and dichlorodihydrofluorescein diacetate (DCF-DA) assay. RESULTS The ethanolic extract of Gynura procumbens inhibited MMP-1 expression up to 70% compare to negative control group. The enzymatic activity of MMP-9 was inhibited around 73% by the treatment of 20μg/mL of the extract. The extract markedly reduced the production of reactive oxygen species (ROS). Gynura procumbens extract showed an inhibitory effect on releasing pro-inflammatory cytokines (IL-6 and IL-8) in human HaCat keratinocyte. CONCLUSION The ethanolic extract of Gynura procumbens inhibited MMP-1 and MMP-9 expressions induced by UV-B irradiation via inhibition of pro-inflammatory cytokine mediator release and ROS production.

Identification of marker compounds in herbal drugs on TLC with DART-MS

This study was conducted to provide a more versatile and specific information on Thin Layer Chromatographic (TLC) analysis of medicinal plants. TLC plates developed with the extract of herbal medicines were analyzed with direct analysis in real time (DART) ion source. Three well known herbal drugs were extracted and developed on a silica-coated TLC plate with the conditions pre-established in Korean Pharmacopoeia IX. The developed plate was placed between the DART ion source and TOF-MS analyzer to get real time mass spectra from the bands on the TLC plate directly. The marker coumarin compounds, decursin and decursinol were successfully identified from the TLC plate developed with Angelicae gigantis radix, along with alkaloid compounds of rutaecarpine and evodiamine from Evodiae fructus, and lignan molecules of gomisin A, N, and schisandrin from Schisandrae fructus. This hyphenation system of TLC and DART-MS could provide unique and specific information on the major constituents of crude plant drug on TLC through uncovering high resolution mass number of each band on the TLC plate directly in real time.

Cognitive-enhancing activity of loganin isolated from Cornus officinalis in scopolamine-induced amnesic mice.

We examined anti-amnesic activity of the methanolic extract of Cornus officinalis fruits (COT) and a major constituent, loganin using scopolamine-induced (1 mg/kg body weight, s.c.) amnesic mice with both passive avoidance and the Morris water maze tests. Oral treatment of mice with COT (100 mg/kg body weight) and loganin (1 and 2 mg/kg body weight) significantly mitigated scopolamine-induced memory deficits in passive avoidance test. In the Morris water maze test, oral treatment of loganin significantly ameliorated scopolamine-induced memory deficits showing the formation of long-term and/or short-term spatial memory. Moreover, loganin (2 mg/kg body weight) significantly inhibited acetylcholinesterase activity by as much as 45% of control in the mouse hippocampus. These results indicate that loganin may exert antiamnesic activity in in vivo through acetylcholinesterase inhibition.

Extract of Salvia miltiorrhiza (Danshen) induces Nrf2-mediated heme oxygenase-1 expression as a cytoprotective action in RAW 264.7 macrophages

ETHNOPHARMACOLOGICAL RELEVANCE Danshen (Salvia miltiorrhiza) is widely used in traditional herbal medicines for relief of a variety of symptoms related to complications arising from vascular diseases such as hypertension, diabetes, and atherosclerosis. Induction of heme oxygenase-1 (HO-1) expression protects against oxidative stress-induced cell damage, which plays an important role in cytoprotection in a variety of pathological models. MATERIALS AND METHODS In the present study, we investigated the effect of Danshen on the up-regulation of HO-1, an inducible and cytoprotective enzyme in RAW 264.7 macrophages. Molecular mechanisms underlying the effects, especially protective effects, was elucidated by analyzing the activation of transcription factors and their upstream signalling, and by evaluating the inhibitory effect of HO-1 on ROS production. RESULTS Danshen induced HO-1 mRNA expression and protein production, and nuclear translocation of NF-E2-related factor 2 in RAW 264.7 macrophages. Pharmacological inhibitors of PI3K/Akt and MEK1 attenuated HO-1 induction in Danshen-stimulated RAW 264.7 macrophages. Furthermore, Danshen pretreatment reduced intracellular production of reactive oxygen species after stimulation with hydrogen peroxide; this effect was reversed by the HO-1 inhibitor ZnPP. CONCLUSION Danshen induced HO-1 expression through PI3K/Akt-MEK1-Nrf2 pathway and reduced intracellular production of reactive oxygen species via induction of HO-1 expression. The results support a role of HO-1 in the cytoprotective effect of Danshen.

Lilium lancifolium Thunb. extract attenuates pulmonary inflammation and air space enlargement in a cigarette smoke-exposed mouse model

ETHNOPHARMACOLOGICAL RELEVANCE Lilium lancifolium Thunb. (Liliaceae) has long been used as a traditional medicine in Korea and China to treat bronchitis, pneumonia, and other pulmonary ailments. AIM OF THE STUDY Cigarette smoke (CS) is a major risk factor for the development of pulmonary inflammatory response; it also triggers pulmonary alveoli enlargement. In the present study, we investigate the effects of Lilium lancifolium Thunb. root extract on pulmonary inflammatory responses in a CS-exposed mouse model. MATERIALS AND METHODS Water extract of Lilium lancifolium Thunb. root was fed to C57BL/6 mice prior CS exposure every day for 3 weeks. The numbers of macrophages and neutrophils in bronchoalveolar lavage fluid (BALF) were counted. The relative inflammatory factors, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), monocyte chemotactic protein-1 (MCP-1), and matrix metalloproteinase-12 (MMP-12) were measured by real-time PCR, ELISA, or Western blot analysis. The average alveoli size was determined by lung histology. RESULTS Lilium lancifolium Thunb. root extract was found to significantly inhibit the numbers of macrophages and neutrophils in BALF due to CS exposure. Lilium lancifolium Thunb. root extract also reduced the protein secretion levels of TNF-α, IL-6, IL-1β, and MCP-1 in BALF and the RNA expression levels of TNF-α, IL-6, IL-1β, MCP-1, and MMP-12 in lung tissue compared with mice only exposed to CS. Moreover, MMP-12 in serum was down regulated in Lilium lancifolium Thunb. root extract treated mice compared with CS-exposed mice. Finally, a morphometric analysis of the lungs of Lilium lancifolium Thunb. root extract treated mice demonstrated a significant reduction in airspace size compared to mice only exposed to CS. CONCLUSION Our results show that Lilium lancifolium Thunb. root extract reduces lung inflammation and airspace enlargement in a CS-exposed mouse model. These data indicate that Lilium lancifolium Thunb. root extract is a therapeutic candidate for pulmonary inflammation and emphysema caused by CS.

Identification of ambiguous cubeb fruit by DART-MS-based fingerprinting combined with principal component analysis.

The cubeb berry has been used as a spice and medicinal herb since medieval times. In China, Litsea cubeba (Lauraceae) has been designated as the cubeb tree; in other countries, Piper cubeba (Piperaceae) is considered as the cubeb. Since most cubeb berries in the Korean market are imported from China, there is a need to develop an efficient analytical method for the identification of the origin of cubeb fruit. Thirty-two commercial cubeb samples were collected from China, Korea, and Indonesia, and direct analysis in real time mass spectrometry (DART-MS) was employed to obtain a chemical fingerprint of each sample for multivariate analysis. Cubebene was identified as a marker molecule for P. cubeba, and all cubeb samples from Korean markets were identified as the fruit of L. cubeba. DART-MS fingerprinting together with principal component analysis can be applied as a promising method for rapid identification and/or quality control of cubeb berries.

Chemometric Classification of Morphologically Similar Umbelliferae Medicinal Herbs by DART-TOF-MS Fingerprint

INTRODUCTION It needs many years of special training to gain expertise on the organoleptic classification of botanical raw materials and, even for those experts, discrimination among Umbelliferae medicinal herbs remains an intricate challenge due to their morphological similarity. OBJECTIVE To develop a new chemometric classification method using a direct analysis in real time-time of flight-mass spectrometry (DART-TOF-MS) fingerprinting for Umbelliferae medicinal herbs and to provide a platform for its application to the discrimination of other herbal medicines. METHODOLOGY Angelica tenuissima, Angelica gigas, Angelica dahurica and Cnidium officinale were chosen for this study and ten samples of each species were purchased from various Korean markets. DART-TOF-MS was employed on powdered raw materials to obtain a chemical fingerprint of each sample and the orthogonal partial-least squares method in discriminant analysis (OPLS-DA) was used for multivariate analysis. RESULTS All samples of collected species were successfully discriminated from each other according to their characteristic DART-TOF-MS fingerprint. Decursin (or decursinol angelate) and byakangelicol were identified as marker molecules for Angelica gigas and A. dahurica, respectively. Using the OPLS method for discriminant analysis, Angelica tenuissima and Cnidium officinale were clearly separated into two groups. Angelica tenuissima was characterised by the presence of ligustilide and unidentified molecular ions of m/z 239 and 283, while senkyunolide A together with signals with m/z 387 and 389 were the marker compounds for Cnidium officinale. CONCLUSION Elaborating with chemoinformatics, DART-TOF-MS fingerprinting with chemoinformatic tools results in a powerful method for the classification of morphologically similar Umbelliferae medicinal herbs and quality control of medicinal herbal products, including the extracts of these crude drugs.

Quantitative analysis of major dibenzocyclooctane lignans in schisandrae fructus by online TLC‐DART‐MS

INTRODUCTION Direct analysis in real time (DART) ion source is a powerful ionising technique for the quick and easy detection of various organic molecules without any sample preparation steps, but the lack of quantitation capacity limits its extensive use in the field of phytochemical analysis. OBJECTIVE To improvise a new system which utilize DART-MS as a hyphenated detector for quantitation. METHODOLOGY A total extract of Schisandra chinensis fruit was analyzed on a TLC plate and three major lignan compounds were quantitated by three different methods of UV densitometry, TLC-DART-MS and HPLC-UV to compare the efficiency of each method. To introduce the TLC plate into the DART ion source at a constant velocity, a syringe pump was employed. The DART-MS total ion current chromatogram was recorded for the entire TLC plate. The concentration of each lignan compound was calculated from the calibration curve established with standard compound. RESULTS Gomisin A, gomisin N and schisandrin were well separated on a silica-coated TLC plate and the specific ion current chromatograms were successfully acquired from the TLC-DART-MS system. The TLC-DART-MS system for the quantitation of natural products showed better linearity and specificity than TLC densitometry, and consumed less time and solvent than conventional HPLC method. CONCLUSION A hyphenated system for the quantitation of phytochemicals from crude herbal drugs was successfully established. This system was shown to have a powerful analytical capacity for the prompt and efficient quantitation of natural products from crude drugs.

Antiobesity Effect of Codonopsis lanceolata in High-Calorie/High-Fat-Diet-Induced Obese Rats.

The antiobesity effects of Codonopsis lanceolata (CL) were evaluated in a high-calorie/high-fat-diet (HFD-) induced obesity rat model and 3T3-L1 cells. The Sprague-Dawley male rats were fed a normal diet (ND) or a HFD for a period of 12 weeks. The rats were subdivided into groups: ND, ND + wild Codonopsis lanceolata (wCL) (900 mg/kg/day, p.o.), ND + cultivated Codonopsis lanceolata (cCL) (900 mg/kg/day, p.o.), HFD, HFD + wCL (100, 300, or 900 mg/kg/day, p.o.), HFD + cCL (100, 300, or 900 mg/kg/day, p.o.), and HFD + sibutramine. The body weight gains of the administered HFD + CL (wCL or CCL) were lower than those of the rats fed with only the HFD group. Moreover, the weight of adipose pads and the serum levels of triglycerides, total cholesterol, and low density lipoprotein cholesterol in the group administered HDL + CL were significantly lower than in the HFD group. The inhibitory effect of lipid accumulation in 3T3-L1 cells was measured by Oil Red O staining and reverse transcription-polymerase chain reaction (RT-PCR). Treatment of 3T3-L1 cells with wCL inhibited lipid accumulation and expression of C/EBPα and PPARγ. These results suggest that CL has a great potential as a functional food with anti-obesity effects and as a therapeutic alternative in the treatment of obesity.