• Title/Summary/Keyword: solid-phase

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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선)

  • Cho, Yoon-Jae;Chae, Young-Sik;Kim, Jae-Young;Kim, Hyung-Soo;Kang, Ilhyun;Do, Jung-Ah;Oh, Jae-Ho;Kwon, Kisung;Chang, Moon-Ik
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.25-33
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    • 2013
  • Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

Identification of Fatty Acids in the Pulp Oils of Jujube and Their Compsitional Changes in the Ripening Period (대추의 과육지질(果肉脂質)에 존재(存在)하는 지방산(脂肪酸)의 동정(同定)과 숙성(熟成)에 따른 그 조성(組成)의 변화(變化))

  • Woo, Hyo-Kyeng;Kim, Seong-Jin;Park, Sung-Hea;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
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    • v.18 no.1
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    • pp.67-77
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    • 2001
  • In search for several fatty acid with unusual structure in vegetable oils, we have found that unknown peaks were shown on GLC in the analysis of fatty acids of the lipids from the pulp of ripened jujube (Zizypus jujuba var. inermis) fruits. These fatty acids were identified as a series of cis-monoenoic acids with ${\omega}-5$ double bond system such as $C_{14:1{\omega}5}$, $C_{16:1{\omega}5}$ and $C_{18:1{\omega}5}$, including ${\omega}-7$ fatty acid as $C_{16:1{\omega}7}$ and $C_{18:1{\omega}7}$, by GLC, solid-phase extraction silver ion-column chromatographic, GLC-mass spectrometric and IR techniques. First of all, total fatty acid methyl esters were resolved into saturated and branched fatty acid, monoenoic acid, dienoic acid, and trienoic acid fraction, respectively, with 100% dichloromethane (DCM), DCM/acetone (9:1, v/v) 100% acetone, and acetone/ acetonitrile (97:3, v/v) solvent system. Unknown fatty acids were included in the monoenoic fraction and were confirmed to have cis-configuration by IR. Picolinyl esters of monoenoic fatty acids gave distinct molecular ion peak and dominant diagnostic peaks, for example, m/z 317, 220 and 260 fragment for $cis-C_{14:1{\omega}5}$, m/z 345, m/z 248 and 288 fragment for $cis-C_{16:1{\omega}5}$ and m/z 373, m/z 276 and 316 fragment for $cis-C_{18:1{\omega}5}$. In this way the occurrence of $cis-C_{16:1{\omega}7}$ and $cis-C_{18:1{\omega}7}$ could be deduced from the appearance of prominent fragments as m/z 345, 220 and 260, and m/z 373, 248 and 280. Level of total ${\omega}-5$ fatty acids amounted to about 30% in the fatty acid composition with the predominance of $C_{16:1{\omega}5}$ $ (18.7{\sim}25.0%)$, in the semi-ripened and/or ripened samples collected in September 14 ($C_{16:1{\omega}5}$ ; 18.7%, $C_{14:1{\omega}5}$ ; 3.6% and $C_{18:1{\omega}5}$ ; 3.0%), September 22 ($C_{16:1{\omega}5}$ ; 25.0%, $C_{14:1{\omega}5}$ ; 1.4% and $C_{18:1{\omega}5}$ ; 2.6%), and October $7 (C_{16:1{\omega}5}$ ; 24.7%, $C_{14:1{\omega}5}$ ; 7.7% and $C_{18:1{\omega}5}$ ; 2.5%). However, the lipids extracted from unripened jujube in July and August contain these unusual fatty acids as low as negligible. It could be observed that the level of ${\omega}-5$ fatty acids in the pulps increased sharply with an elapse of ripening time of jujube fruits. Other monoenoic fatty acids with ${\omega}-7$ series, $C_{16:1{\omega}7}$ (palmitoleic acid) and $C_{18:1{\omega}7}$ (cis-vaccenic acid) could be detected. And in the lipids of the kernel and leaf of jujube, none of ${\omega}-5$ fatty acids could be detected.

Improvement of Radiosynthesis Yield of [11C]acetate ([11C]아세트산의 방사화학적 수율 증가를 위한 연구)

  • Park, Jun Young;Son, Jeongmin
    • The Korean Journal of Nuclear Medicine Technology
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    • v.22 no.2
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    • pp.74-78
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    • 2018
  • Purpose $[^{11}C]$acetate has been proved useful in detecting the myocardial oxygen metabolism and various malignancies including prostate cancer, hepatocellular carcinoma, renal cell carcinoma and brain tumors. The purpose of study was to improve the radiosynthesis yield of $[^{11}C]$acetate on a automated radiosynthesis module. Materials and Methods $[^{11}C]$acetate was prepared by carboxylation of grignard reagent, methylmagnesium chloride, with $[^{11}C]$$CO_2$ gas, followed by hydrolysis with 1 mM acetic acid and purification using solid phase extraction cartridges. The effect of the reaction temperature ($0^{\circ}C$, $10^{\circ}C$, $-55^{\circ}C$) and cyclotron beam time (10 min, 15 min, 20 min, 25 min) on the radiosynthesis yield were investigated in the $[^{11}C]$acetate labeling reaction. Results The maximum radiosynthesis yield was obtained at $-10^{\circ}C$ of reaction temperature. The radioactivities of $[^{11}C]$acetate acquired at $-10^{\circ}C$ reaction temperature was 2.4 times higher than those of $[^{11}C]$acetate acquired at $-55^{\circ}C$. Radiosynthesis yield of $[^{11}C]$acetate increased with increasing cyclotron beam time. Conclusion This study shows that radiosynthesis yield of $[^{11}C]$acetate highly dependent on reaction temperature. The best radiosynthesis yield was obtained in reaction of grignard reagent with $[^{11}C]$$CO_2$ at $-10^{\circ}C$. This radiolabeling conditions will be ideal for routine clinical application.

Development and Validation of an Analytical Method for Quinoxyfen in Agricultural Products using QuEChERS and LC-MS/MS (QuEChERS법 및 LC-MS/MS를 이용한 농산물 중 살균제 Quinoxyfen의 잔류시험법 개발 및 검증)

  • Cho, Sung Min;Do, Jung-Ah;Lee, Han Sol;Park, Ji-Su;Shin, Hye-Sun;Jang, Dong Eun;Choi, Young-Nae;Jung, Yong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.2
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    • pp.140-147
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    • 2019
  • An analytical method was developed for the determination of quinoxyfen in agricultural products using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted with 1% acetic acid in acetonitrile and water was removed by liquid-liquid partitioning with $MgSO_4$ (anhydrous magnesium sulfate) and sodium acetate. Dispersive solid-phase extraction (d-SPE) cleanup was carried out using $MgSO_4$, PSA (primary secondary amine), $C_{18}$ (octadecyl) and GCB (graphitized carbon black). The analytes were quantified and confirmed by using LC-MS/MS in positive mode with MRM (multiple reaction monitoring). The matrix-matched calibration curves were constructed using six levels ($0.001-0.25{\mu}g/mL$) and the coefficient of determination ($R^2$) was above 0.99. Recovery results at three concentrations (LOQ, 10 LOQ, and 50 LOQ, n=5) were in the range of 73.5-86.7% with RSDs (relative standard deviations) of less than 8.9%. For inter-laboratory validation, the average recovery was 77.2-95.4% and the CV (coefficient of variation) was below 14.5%. All results were consistent with the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and Food Safety Evaluation Department guidelines (2016). The proposed analytical method was accurate, effective and sensitive for quinoxyfen determination in agricultural commodities. This study could be useful for the safe management of quinoxyfen residues in agricultural products.

Development and Validation of an Analytical Method for Fenpropimorph in Agricultural Products Using QuEChERS and LC-MS/MS (QuEChERS법과 LC-MS/MS를 이용한 농산물 중 Fenpropimorph 시험법 개발 및 검증)

  • Lee, Han Sol;Do, Jung-Ah;Park, Ji-Su;Cho, Sung Min;Shin, Hye-Sun;Jang, Dong Eun;Choi, Young-Nae;Jung, Yong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.2
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    • pp.115-123
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    • 2019
  • An analytical method was developed for the determination of fenpropimorph, a morpholine fungicide, in hulled rice, potato, soybean, mandarin and green pepper using QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) sample preparation and LC-MS/MS (liquid chromatography-tandem mass spectrometry). The QuEChERS extraction was performed with acetonitrile followed by addition of anhydrous magnesium sulfate and sodium chloride. After centrifugation, d-SPE (dispersive solid phase extraction) cleanup was conducted using anhydrous magnesium sulfate, primary secondary amine sorbents and graphitized carbon black. The matrix-matched calibration curves were constructed using seven concentration levels, from 0.0025 to 0.25 mg/kg, and their correlation coefficient ($R^2$) of five agricultural products were higher than 0.9899. The limits of detection (LOD) and quantification (LOQ) were 0.001 and 0.0025 mg/kg, respectively, and the limits of quantification for the analytical method were 0.01 mg/kg. Average recoveries spiked at three levels (LOQ, $LOQ{\times}10$, $LOQ{\times}50$, n=5) and were in the range of 90.9~110.5% with associated relative standard deviation values less than 5.7%. As a result of the inter-laboratory validation, the average recoveries between the two laboratories were 88.6~101.4% and the coefficient of variation was also below 15%. All optimized results were satisfied the criteria ranges requested in the Codex guidelines and Food Safety Evaluation Department guidelines. This study could serve as a reference for safety management relative to fenpropimorph residues in imported and domestic agricultural products.

Comparative Study on the Composition of Floral Volatile Components in the Flowering Stages of Robinia pseudoacacia L. (아까시나무(Robinia pseudoacacia L.) 꽃의 개화 단계별 향기성분 조성 비교)

  • Jung, Je Won;Lee, Hyun Sook;Noh, Gwang Rae;Lee, Andosung;Kim, Moon Sup;Kim, Sea Hyun;Kwon, Hyung Wook
    • Journal of Apiculture
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    • v.32 no.3
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    • pp.139-146
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    • 2017
  • Floral scent emitted from many plants is the critical factors for pollinator attraction and defense for adaptation in environments. The fragrance components of flowers are different in composition by geographical origins, climate factors and the development stages of flowers. In the present study, we investigated the volatile-floral compounds in flowers of Robinia pseudoacacia L. and defined the chemical contribution for flowering periods. The volatile compounds analysis was performed by gas chromatography with mass selective detector after solid phase microextraction (SPME). We reported different compositional features of fragrance compounds according to flowering periods. The abundant compounds identified in stage 1 were ${\alpha}$-pinene (66.80%) and ${\beta}$-pinene (26.53%). Those of the stage 2 were (Z)-${\beta}$-ocimene (37.57%), ${\alpha}$-pinene (15.16%), benzaldehyde (16.63%), linalool (12.13%). The volatiles of stage 3 comprised an abundance of (Z)-${\beta}$-ocimene (64.94%), ${\alpha}$-pinene (9.84%), linalool (8.92%), benzaldehyde (1.71%). Leaf volatiles were distinct from those in the reproductive plant parts by their high relative amount of (E)-${\beta}$-ocimene (23.50%) and (Z)-3-Hexenyl acetate (27.87%). Differences in flower scents of the different stages and leaves are discussed in light of biochemical constraints on volatile chemical synthesis and of the role of flower scent in evolutionary ecology of R. pseudoacacia.

Evaluation of the Potential of Nitrogen Plasma to Cosmetics (질소 플라즈마의 화장품 가능성 평가)

  • Lee, So Min;Jung, So Young;Brito, Sofia;Heo, Hyojin;Cha, Byungsun;Lei, Lei;Lee, Sang Hun;Lee, Mi-Gi;Bin, Bum-Ho;Kwak, Byeong-Mun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.3
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    • pp.189-196
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    • 2022
  • Plasma refers to an ionized gas that is often referred to as "the fourth phase of matter", following solid, liquid, and gas. Plasma has traditionally been utilized for industrial applications such as welding and neon signs, but its promise in biomedical fields such as cancer treatment and dermatology has lately been recognized. Indeed, due to its beneficial effects in promoting collagen production, improving skin tone, and eliminating harmful bacteria in the skin, plasma treatment constitutes an important target for dermatological research. In this study, a plasma device for cosmetic manufacturing based on nitrogen, the main component of the atmosphere, was designed and assembled. Moreover, nitric oxide (NO) was selected since is easier to follow and evaluate than other nitrogen plasma active species, and its contents were measured to perform a quantitative and qualitative evaluation of plasma. First, an injection method, using different proximities labeled "sinking" and "non sinking" treatments, was performed to test the most efficient plasma treatment method. As a result, it was observed that the formulation obtained by a non sinking treatment was more effective. Furthermore, toner and ampoule were selected as cosmetics formulations, and the characteristics of the formulation and changes in the injected plasma state were observed. In both formulations, the successful injection of NO plasma was 2 times higher in toner formulation than ampoule formulation, and it gradually decreased with time, having dissipated after a week. It was confirmed that the nitrogen plasma used did not affect the stability of the toner and ampoule formulations at low temperature (4 ℃), room temperature (25 ℃), and high temperature (37 ℃ and 50 ℃) conditions. The results of this study demonstrate the potential of plasma cosmetics and highlight the importance of securing the stability of the injected plasma.

A Study on the Recovery of Lithium from Secondary Resources of Ceramic Glass Containing Li-Al-Si by Ca-based Salt Roasting and Water Leaching Process (Li-Al-Si 함유 유리세라믹 순환자원으로부터 Ca계열 염배소법 및 이에 따른 수침출 공정에 의한 리튬의 회수 연구)

  • Sung-Ho Joo;Dong Ju Shin;Dongseok Lee;Shun Myung Shin
    • Resources Recycling
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    • v.32 no.1
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    • pp.42-49
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    • 2023
  • The glass ceramic secondary resource containing Li-Al-Si is used in inductor, fireproof glass, and transparent cookware and accounts for 14% of the total consumption of Li, which is the second most widely used after Li-ion batteries. Therefore, new Li resources should be explored when the demand for Li is exploding, and extensive research on Li recovery is needed. Herein, we recovered Li from fireproof Li-Al-Si glass ceramic, which is a new secondary resource containing Li. The fireproof glass among all Li-Al-Si glass ceramics was used as raw material that contained 1.5% Li, 9.4% Al, and 28.9% Si. The process for recovering Li from the fireproof glass was divided into two parts: (1) calcium salt roasting and (2) water leaching. In calcium salt roasting, a sample of fireproof glass was crushed and ground below 325 mesh. The leaching efficiency was compared based on the presence or absence of heat treatment of the fireproof glass. Moreover, the leaching rates based on the input ratios of calcium salt, Li-Al-Si glass, and ceramics and the leaching process based on calcium salt roasting temperatures were compared. In water leaching, the leaching and recovery rates of Li based on different temperatures, times, solid-liquid ratios, and number of continuous leaching stages were compared. The results revealed that fireproof glass ceramics containing Li-Al-Si should be heat treated to change phase to beta-type spodumene. CaCO3 salt should be added at a ratio of 6:1 with glass ceramics containing Li-Al-Si, and then leached 4 times or more to achieve a recovery efficiency of Li over 98% from a solution containing 200 mg/L of Li.

A Case Study of Shanghai Tang: How to Build a Chinese Luxury Brand

  • Heine, Klaus;Phan, Michel
    • Asia Marketing Journal
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    • v.15 no.1
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    • pp.1-22
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    • 2013
  • This case focuses on Shanghai Tang, the first truly Chinese luxury brand that appeals to both Westerners and, more recently, to Chinese consumers worldwide. A visionary and wealthy businessman Sir David Tang created this company from scratch in 1994 in Hong Kong. Its story, spanned over almost two decades, has been fascinating. It went from what best a Chinese brand could be in the eyes of Westerners who love the Chinese culture, to a nearly-bankrupted company in 1998, before being acquired by Richemont, the second largest luxury group in the world. Since then, its turnaround has been spectacular with a growing appeal among Chinese luxury consumers who represent the core segment of the luxury industry today. The main objective of this case study is to formally examine how Shanghai Tang overcame its downfall and re-emerged as one the very few well- known Chinese luxury brands. More specifically, this case highlights the ways with which Shanghai Tang made a transitional change from a brand for Westerners who love the Chinese culture, to a brand for both, Westerners who love the Chinese culture and Chinese who love luxury. A close examination reveals that Shanghai Tang has followed the brand identity concept that consists of two major components: functional and emotional. The functional component for developing a luxury brand concerns all product characteristics that will make a product 'luxurious' in the eyes of the consumer, such as premium quality of cachemire from Mongolia, Chinese silk, lacquer, finest leather, porcelain, and jade in the case of Shanghai Tang. The emotional component consists of non-functional symbolic meanings of a brand. The symbolic meaning marks the major difference between a premium and a luxury brand. In the case of Shanghai Tang, its symbolic meaning refers to the Chinese culture and the brand aims to represent the best of Chinese traditions and establish itself as "the ambassador of modern Chinese style". It touches the Chinese heritage and emotions. Shanghai Tang has reinvented the modern Chinese chic by drawing back to the stylish decadence of Shanghai in the 1930s, which was then called the "Paris of the East", and this is where the brand finds inspiration to create its own myth. Once the functional and emotional components assured, Shanghai Tang has gone through a four-stage development to become the first global Chinese luxury brand: introduction, deepening, expansion, and revitalization. Introduction: David Tang discovered a market gap and had a vision to launch the first Chinese luxury brand to the world. The key success drivers for the introduction and management of a Chinese luxury brand are a solid brand identity and, above all, a creative mind, an inspired person. This was David Tang then, and this is now Raphael Le Masne de Chermont, the current Executive Chairman. Shanghai Tang combines Chinese and Western elements, which it finds to be the most sustainable platform for drawing consumers. Deepening: A major objective of the next phase is to become recognized as a luxury brand and a fashion or design authority. For this purpose, Shanghai Tang has cooperated with other well-regarded luxury and lifestyle brands such as Puma and Swarovski. It also expanded its product lines from high-end custom-made garments to music CDs and restaurant. Expansion: After the opening of his first store in Hong Kong in 1994, David Tang went on to open his second store in New York City three years later. However this New York retail operation was a financial disaster. Barely nineteen months after the opening, the store was shut down and quietly relocated to a cheaper location of Madison Avenue. Despite this failure, Shanghai Tang products found numerous followers especially among Western tourists and became "souvenir-like" must-haves. However, despite its strong brand DNA, the brand did not generate enough repeated sales and over the years the company cumulated heavy debts and became unprofitable. Revitalizing: After its purchase by Richemont in 1998, Le Masne de Chermont was appointed to lead the company, reposition the brand and undertake some major strategic changes such as revising the "Shanghai Tang" designs to appeal not only to Westerners but also to Chinese consumers, and to open new stores around the world. Since then, Shanghai Tang has become synonymous to a modern Chinese luxury lifestyle brand.

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