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Characterizing a full spectrum of physico-chemical properties of (20S)-and (20R)-ginsenoside Rg3 to be proposed as standard reference materials

  • Kim, Il-Woung;Sun, Won Suk;Yun, Bong-Sik;Kim, Na-Ri;Min, Dongsun;Kim, Si-Kwan
    • Journal of Ginseng Research
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    • v.37 no.1
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    • pp.124-134
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    • 2013
  • The authentication of the physico-chemical properties of ginsenosides reference materials as well as qualitative and quantitative batch analytical data based on validated analytical procedures is a prerequisite for certifying good manufacturing practice (GMP). Ginsenoside Rb1 and Rg1, representing protopanaxadiol and protopanaxatriol ginsenosides, respectively, are accepted as marker substances in quality control standards worldwide. However, the current analytical methods for these two compounds recommended by Korean, Chinese, European, and Japanese pharmacopoeia do not apply to red ginseng preparations, particularly the extract, because of the relatively low content of the two agents in red ginseng compared to white ginseng. In manufacturing fresh ginseng into red ginseng products, ginseng roots are exposed to a high temperature for many hours, and the naturally occurring ginsenoside Rb1 and Rg1 are converted to artifact ginsenosides such as Rg3, Rg5, Rh1, and Rh2 during the heating process. The analysis of ginsenosides in commercially available ginseng products in Korea led us to propose the inclusion of the (20S)- and (20R)-ginsenoside Rg3, including ginsenoside Rb1 and Rg1, as additional reference materials for ginseng preparations. (20S)- and (20R)-ginsenoside Rg3 were isolated by Diaion HP-20 adsorption chromatography, silica gel flash chromatography, recrystallization, and preparative HPLC. HPLC fractions corresponding to those two ginsenosides were recrystallized in appropriate solvents for the analysis of physico-chemical properties. Documentation of those isolated ginsenosides was achieved according to the method proposed by Gaedcke and Steinhoff. The ginsenosides were subjected to analyses of their general characteristics, identification, purity, content quantification, and mass balance tests. The isolated ginsenosides showed 100% purity when determined by the three HPLC systems. Also, the water content was found to be 0.534% for (20S)-Rg3 and 0.920% for (20R)-Rg3, meaning that the net mass balances for (20S)-Rg3 and (20R)-Rg3 were 99.466% and 99.080%, respectively. From these results, we could assess and propose a full spectrum of physico-chemical properties of (20S)- and (20R)-ginsenoside Rg3 as standard reference materials for GMP-based quality control.

Isolation and Identification of Antioxidant Substances from the Stems of Butterbur (Petasites japonicus) (머위(Petasites japonicus) 엽병으로부터 항산화 물질의 분리 및 동정)

  • Kim, Min-Young;Yi, Jung-Hyun;Hwang, Yun-Yi;Song, Kyung-Sik;Jun, Mi-Ra
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.8
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    • pp.979-984
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    • 2008
  • The stems of P etasites japonicus were extracted with ethanol and then partitioned with hexane, chloroform, ethyl acetate, n-butanol and water, successively. The antioxidant potency of five crude fractions were determined using (1) 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, (2) thiobarbituric acid reactive substances (TBARS) assay in the linoleic acid model system, and (3) lipoxygenase inhibition assay. Among the crude fractions, the ethyl acetate fraction exhibited the most potent antioxidant effect. By activity-guided fractionation, compound PJ-4 was isolated from the ethyl acetate fraction through the repeated silica gel open column chromatography. The chemical structure of the isolated compound was determined as kaempferol by $^1H-$ and $^{13}C$-NMR analysis and its antioxidative capacity was further investigated. DPPH radical scavenging activity of the compound was 65.76% at the concentration of $100 \;{\mu}g/mL$. The inhibitory activity of the compound against lipid peroxidation and lipoxygenase exhibited 43.47% and 58.60%, respectively at the concentration of $100\;{\mu}g/mL$. The result suggests that the compound may serve as a useful natural antioxidant and furthermore indicates the possibility of developing the stems of Petasites japonicus as a natural antioxidant source.

Influence of Ionic Liquid for Separation of D-tryptophan and N-CBZ-D-phenylalanine (D-tryptophan과 N-CBZ-D-phenylalanine의 분리에서 이온성 액체의 영향)

  • Jin, Yinzhe;Zheng, Jinzhu;Polyakova, Yulia;Koo, Yoon Mo;Row, Kyung Ho
    • Korean Chemical Engineering Research
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    • v.44 no.5
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    • pp.453-459
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    • 2006
  • D-tryptophan and N-CBZ-D-phenylalanine were separated using ionic liquid as additives for the mobile phase in high performance liquid chromatography (HPLC). The ionic liquid of 1-butyl-3- methylimidazolium tetrafluoroborate (${[BMIm]}^+{[BF_4]}^-$) was used. Mobile phases were 65%, 70%, and 80% methanol in water with addition of different concentrations (0.5, 1.0, 1.5, 2.0, 4.0, 6.0, 8.0, 10.0, 12.0, and 15.0 mmol/L) of the ionic liquid. The experiments were performed on stainless steel column, $3.9{\times}300mm$ i.d., packed with $15{\mu}m$ octadecyl-bonded silica gel at laboratory.The retention factor of D-tryptophan was not negligibly changed while that of N-CBZ-D-phenylalanine was decreased. The resolution between the two components were affected by the contents of methanol and ionic liquid in the mobile phase. With the small content of methanol and the high concentration of ionic liquid, the resolution was improved.

Studies on the Cellulase. (V) -Fractionation of Cellulolytic Complex produced by Trichoderma $viride(O_2-1)$ (섬유소(纖維素) 분해효소(分解酵素)에 관(關)한 연구(硏究) (제5보(第5報)) -Trichoderma $(O_2-1)$가 생성(生成)하는 Cellulolytic Complex의 분별(分別)에 대(對)하여-)

  • Sung, Nack-Kie
    • Applied Biological Chemistry
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    • v.12
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    • pp.99-105
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    • 1969
  • The yield of cellulase derived from Trichoderma $(O_2-1)$ was remarkably varied with various concentration of ethanol and acetone in purification of the enzyme. In the purification with ethanol of ${\beta}-glucosidase$, the best result was obtained in the concentration of 60% and, of CMCase and of filter paper disintegrating enzyme 80%. And in the purification with acetone of ${\beta}-glucosidase$, filter paper disintegrating enzyme, and CMCase, in the concentration of 60%, 80%, and 90% respectively, was shown the best yield. The activities of crude Cellulase preparation could be seperated into few of fractions by column chromatography with Silica gel, Cellulose powder, and gauze. Most of CMCase, avicelase, and ${\beta}-glucosidase$ were eluted, but most of filter paper disintegrating enzyme and the rest of enzymes mentioned the above were absorbed, and were eluted with water. Therefore, it was considered that CMCase is different from filter paper disintegrating enzyme in properties. The relative activity of CMCase was different from that of avicelase in the peak of elusion part. And it was considered that filter paper disintegrating enzyme and cellulose powder saccharifying enzyme was seperated respectively as absorption part and non absorption part. The auther came to the conclusion that at least there were more than three sorts of cellulase in Trichoderma $(O_2-1)$ cellulase preparation.

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A Study on Discriminative Criteria of 6 Kinds of Achyranthis Radix Using HPLC/DAD;Isolation and Identification of 20-hydroxyecdysone from Aclryranthes japonica $N-{AKAI}$ and Comparison of Patterns of Achyranthis Radix from Different Locations by HPLC (HPLC/DAD를 이용한 6종(種) 우슬(牛膝)의 분류기준 연구;우슬(牛膝)(쇠무릎, Achyranthes japonica $N_{AKAI}$)로부터 20-hydroxyecdysone 분리.동정 및 산지별 우슬의 HPLC 패턴 비교)

  • Kim, Jeong-Hi;Kim, Jong-Mun;Kang, Dae-Hoon
    • 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.

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A STUDY ON THE PHYSICAL PROPERTIES OF GLASS IONOMER CEMENT FOR RESTORATIVE FILLING USING VISIBLE LIGHT POLYMERIZATION (가시광선중합화에 따른 충전용 Glass Ionomer Cement의 물리적 성질에 관한 연구)

  • Shin, Dong-Hoon;Kwon, Hyuk-Choon
    • Restorative Dentistry and Endodontics
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    • v.17 no.2
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    • pp.307-330
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    • 1992
  • The aim of this study was to investigate the physical properties of visible light curing Glass Ionomer cement for restorative esthetic filling. The control group was the autopolymerizing GC Fuji II Glass Ionomer cement (2.2: 1 P/L ratio) and the experimental groups were made by following procedure. To induce the polymerization by visible light, the powder of GC Fuji II GI cement and the liquid of Vitrabond for base & liner were mixed in an amalgam capsule with 2.5:1, 3.0:1, 3.5:1 P/L ratio (% wt/wt). After fabrication of specimens, compressive strength, fracture toughness ($K_{IC}$) Scanning Electron Microscope and X-ray Diffraction, water-leachable content, marginal leakage and surface roughness were studied. The results were as follows: 1. Only experimental No. 1 group (visible light curing) showed less compressive strength than control group 1 hour after curing. Strength was increased with aging in all groups, so the compressive strength of light curing groups was no less than that of autopolymerizing group after 3 weeks. 2. Experimental No.3 group (visible light curing) was inferior to No.2 group (visible light curing) in fracture resistance but light curing groups were more resistant to fracture than autopolymerizing group and showed ductile fracture pattern as compared with the brittle fracture pattern of autopolymerizing group. 3. From scanning electron microscopic image, various sized unreacted powder particles, surrounded by silica gel, were embedded in polysalt matrix. Light curing groups showed little crack and more dense unreacted particles than autopolymerizing group. 4. From X-ray diffraction analysis, GC Fuji II Glass Ionomer cement powder and all groups showed glassy appearance but light curing groups seemed to be more intensive in crystaline than autopolymerizing group. S. The most significant dissolution was shown in early setting period in all group. Light curing groups were dissolved less than autopolymerizing group. 6. Marginal leakage was not different significantly in case of cavity margin composed of same tooth structure (ex. only enamel margin, only dentin margin) but much more leakage was shown in dentin/cementum margin than enamel margin. In only case of only enamel margin, light curing groups were superior to autopolymerizing group. 7. All groups showed relatively smooth surface, which irregularity was less than $1{\mu}m$. Light curing groups were smoother than autopolymerizing group.

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A Study on the Antitumor Activity of Panax ginseng (고려인삼의 항암효과에 관한 연구)

  • Hwang, Woo-lk
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.52-60
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    • 1993
  • Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration of the crude GX or 7 : 3 GX in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro. In the meantime, ginseng saponin derivatives did not have cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7 : 3 GX was about 3 times more potent than that of crude GX, one unit of cytotoxic activity against L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri value of the active compound on silica- gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90 : 10 : 1, v/v/v) as a developing so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7 : 3 GX treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gt The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude GX, which can explain a part of the origin of its anticancer activity.

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Antioxidant Activity of Different Parts of Lespedeza bicolor and Isolation of Antioxidant Compound (싸리나무(Lespedeza bicolor) 부위별 추출물의 항산화 활성 및 항산화물질 분리)

  • Lee, Jae-Hak;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.763-771
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    • 2012
  • In this study, total antioxidant properties of extracts from different parts of Lespedeza bicolor were determined using techniques of measuring 1,1-diphenyl-2-picryl hydrazyl/2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)-radical scavenging activity and total phenolic contents. The total antioxidant activities of leaf, stem and root extracts from various solvents (water, 50, 70, 100% ethanol, and hot-water) indicated that 50 and 70% ethanol extracts have high radical scavenging activities and phenolic contents. A systematic approach was used to determine the total antioxidant activity of different solvent fractions of the Lespedeza bicolor extracts, partitioning with chloroform, ethyl acetate, n-butanol, and water, and the ethyl acetate fraction was found to have the strongest antioxidant activity. Antioxidant assay-guided isolation was carried out to isolate potential antioxidant compounds. The ethyl acetate fraction of the leaf extract was subjected to silica gel, LH-20 and RP-18 column chromatography successively, and afforded compound 1, which was identified as eriodictyol by NMR and MS analysis, after which its antioxidant activity was determined.

Performance Evaluation of Hydrocyclone Filter for Treatment of Micro Particles in Storm Runoff (Hydrocyclone Filter 장치를 이용한 강우유출수내 미세입자 제거특성 분석)

  • Lee, Jun-Ho;Bang, Ki-Woong;Hong, Sung-Chul
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.11
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    • pp.1007-1018
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    • 2009
  • Hydrocyclone is widely used in industry, because of its simplicity in design, high capacity, low maintenance and operational cost. The separation action of a hydrocyclone treating particulate slurry is a consequence of the swirling flow that produces a centrifugal force on the fluid and suspended particles. In spite of hydrocyclone have many advantage, the application for treatment of urban stormwater case study were rare. We conducted a laboratory scale study on treatable potential of micro particles using hydrocyclone filter (HCF) that was a combined modified hydrocyclone with perlite filter cartridge. Since it was not easy to use actual storm water in the scaled-down hydraulic model investigations, it was necessary to reproduce ranges of particles sizes with synthetic materials. The synthesized storm runoff was made with water and addition of particles; ion exchange resin, road sediment, commercial area manhole sediment, and silica gel particles. Experimental studies have been carried out about the particle separation performance of HCF-open system and HCF-closed system. The principal structural differences of these HCFs are underflow zone structure and vortex finder. HCF was made of acryl resin with 120 mm of diameter hydrocyclone and 250 mm of diameter filter chamber and overall height of 800 mm. To determine the removal efficiency for various influent concentrations of suspended solids (SS) and chemical oxygen demand (COD), tests were performed with different operational conditions. The operated maximum of surface loading rate was about 700 $m^3/m^2$/day for HCF-open system, and 1,200 $m^3/m^2$/day for HCF-closed system. It was found that particle removal efficiency for the HCF-closed system is better than the HCF-open system under same surface loading rate. Results showed that SS removal efficiency with the HCF-closed system improved by about 8~20% compared with HCF-open system. The average removal efficiency difference for HCF-closed system between measurement and CFD particle tracking simulation was about 4%.

Radiolysis Assessment of $^{18}F$-FDG According to Automatic Synthesis Module (자동합성장치에 따른 $^{18}F$-FDG의 방사선분해 평가)

  • Kim, Si-Hwal;Kim, Dong-Il;Chi, Yong-Gi;Choi, Sung-Wook;Choi, Choon-Ki;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.1
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    • pp.8-11
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    • 2012
  • Purpose : Among quality control items, the radiochemical impurity must be below 10% of total radioactivity. In this regard, as the recently commercialized automatic synthesis module produces a large amount of 18F-FDG, radiolysis of radiopharmaceuticals is very likely to occur. Thus, this study compared the changes in radiochemical purity regarding radiolysis of $^{18}F$-FDG according to automatic synthesis module. Materials and methods : Cyclotron (PETtrace, GE Healthcare) was used to produce $^{18}F$ and automatic synthesis module (FASTlab, Tracerlab MX, GE Healthcare) was used to achieve synthesis into FDG. For radiochemical purity, Radio-TLC Scanner (AR 2000, Bioscan), GC (Gas Chromatograph, Agilent 7890A) was used to measure the content of ethanol included in $^{18}F$-FDG. Glass board applied with silica gel ($1{\times}10cm$) was used for stationary phase while a mixed liquid formed of acetonitrile and water (ratio 19:1) was used for mobile phase. High-concentration and low-concentration $^{18}F$-FDG were produced in each synthesis module and the radiochemical purity was measured every 2 hours. Results : The purity in low-concentration (below 2.59 GBq/mL) was measured as 99.26%, 98.69%, 98.25%, 98.09% in Tracerlab MX and as 99.09%, 97.83%, 96.89%, 96.62% in FASTlab according to 0, 2, 4, 6 hours changes, respectively. The purity in high-concentration (above 3.7 GBq/mL) was measured as 99.54%, 96.08%, 93.77%, 92.54% in Tracerlab MX and as 99.53%, 95.65%, 92.39%, 89.82% in FASTlab according to 0, 2, 4, 6 hours changes, respectively. Also, ethanol was not detected in GC of $^{18}F$-FDG produced in FASTlab, while 100~300 ppm ethanol was detected in Tracerlab MX. Conclusion : Whereas the change of radiochemical purity was only 3% in low-concentration $^{18}F$-FDG, the change was rapidly increased to 10% in high-concentration. Also, higher radiolysis were observed in $^{18}F$-FDG produced in FASTlab than Tracerlab MX. This is because ethanol is included in the synthesis stage of Tracerlab MX but not in the synthesis stage of FASTlab. Thus, radiolysis is influenced by radioactivity concentration than the inclusion of ethanol, which is the radioprotector. Therefore, after producing high-concentration $^{18}F$-FDG, the content must be diluted through saline to lower concentration.

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