• Title/Summary/Keyword: Synthesis condition

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Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity (청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가)

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.

New Synthesis of Sestamibi and Comparison of Stability of Its Formulation (Sestamibi의 신규합성과 제제화에 따른 안정성 비교)

  • Son, Mi-Won;Lim, Joong-In;Chang, Young-Soo;Jung, Mi-Young;Jeong, Lak-Shin;Kim, Soon-Hoe;Kim, Won-Bae;Jeong, Jae-Min
    • The Korean Journal of Nuclear Medicine
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    • v.35 no.5
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    • pp.334-341
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    • 2001
  • Purpose: Ascorbic acid us known to act as an antioxidant. Therefore, it can be used in increasing the efficiency of radiochemical labeling of Technetium-99m setamibi by inhibition of oxidation of $Sn^{2+}$ at low concentration. We intended to estimate the efficiency of radiochemical labeling and the stability of the newly formed formulation when ascorbic acid was added to a commercial kit. Materials and Methods: Synthesis of sestamibi was performed according to Dong-A's patent procedure (No.10-2001-0012877). First, we undertook a study to evaluate the efficiency of radiochemical labeling of sestamibi containing ascorbic acid. The stability of the vials was assessed using either $7.5{\mu}g\;or\;75{\mu}g$ of ascorbic acid, added to commercial vials under the accelerated condition(Temp : $40^{\circ}C{\pm}2^{\circ}C$, Relative humidity : $75{\pm}5%$). Results: Sestamibi was synthesized in overall 35-40% yield over 5 steps from a commercially available methallyl chloride as a starling material. When ascorbic acid was added, the efficiency of radiochemical labeling was maintained compared to the vial with no ascorbic acid. The accelerated test showed that the addition of ascorbic acid inhibited the oxidation of $Sn^{2+}$ ion by antioxidation mechanism. Also, the efficiency of radiochemical labeling of this vial after 9 months was nearly the same as the starting point. Therefore, the storage period of the kit is likely to be extended. Taken together, it suggests that the addition of ascorbic acid as a stabilizer is desirable. Conclusion: To increase the stability of a sestamibi cold kit, it is desirable to add ascorbic acid as a stabilizer to the commercial formulation.

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Purification and Characterization of $\beta$-Cyclodextrin Glucanotransferase Excreted by Bacillus firmus var. aikalophilus. (호알칼리성 Bacillus firmus가 생산하는 $\beta$-Cyclodextrin Glucanotransferase의 정제 및 효소반응 특성)

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.323-330
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    • 1998
  • Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0~9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

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Disturbance of $\alpha$-Amylase Secretion from Bacillus amyloliquefaciens Cells by the Treatment of Puromycin and Magnesium (Bacillus amyloliquefaciens에서 Puromycin 과 Magnesium에 의한 $\alpha$-Amylase 의 분비저해)

  • 안순자;김순옥;이동희;송방호
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.412-420
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    • 1989
  • To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.

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EFFECT OF ENAMEL MATRIX DERIVATIVE (EMD, $EMDOGAIN^{(R)}$) ON THE DIFFERENTIATION OF CULTURED HUMAN PERIODONTAL LIGAMENT CELLS AND MESENCHYMAL STEM CELLS (배양된 사람 치주인대세포와 골수유래간엽줄기세포의 분화에 미치는 법랑기질유도체 (Enamel Matrix Derivative, EMD)의 영향)

  • Park, Sang-Gyu;Jue, Seong-Suk;Kwon, Yong-Dae;Choi, Byung-Joon;Kim, Young-Ran;Lee, Baek-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.4
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    • pp.281-286
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    • 2009
  • Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

Effect of LED mixed light conditions on the glucosinolate pathway in brassica rapa (배추 유묘의 글루코시놀레이트 합성 기작에 미치는 LED 혼합광의 효과)

  • Moon, Junghyun;Jeong, Mi Jeong;Lee, Soo In;Lee, Jun Gu;Hwang, Hyunseung;Yu, Jaewoong;Kim, Yong-Rok;Park, Se Won;Kim, Jin A
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.245-256
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    • 2015
  • In the agricultural industries, LEDs are used as supplementary, as well as main lighting sources in closed cultivation systems. In cultivation using artificial light sources, various light qualities have been tried to supplement fluorescent lamps to promote plant growth and metabolism. Microarray analysis of Brassica rapa seedlings under blue and fluorescent mixed with blue light conditions identified changes in three genes of the glucosinolate pathway. This attracted attention as functional materials highly expressed 3.6-4.6 fold under latter condition. We selected four more genes of the glucosinolate pathway from the Brassica database and tested their expression changes under fluorescent light mixed with red, green, and blue, respectively. Some genes increased expression under red and blue mixed conditions. The Bra026058, Bra015379, and Bra021429; the orthologous genes of CYP79F1, ST5a, and FMOGS-OX1 in Arabidopsis, are highly expressed in Brassica rapa under fluorescent mixed with blue light conditions. Further, Bra029355, Bra034180, Bra024634, and Bra022448; the orthologous genes of MAM1, AOP3, UGT74B1, and BCAT4 in Arabidopsis, are highly expressed in Brassica rapa under fluorescent mixed with red light conditions. The various light conditions had unique effects on the varieties of Brassica, resulting in differences in glucosinolate synthesis. However, in some varieties, glucosinolate synthesis increased under mixed blue light conditions. These results will help to construct artificial light facilities, which increase functional crops production.

Dietary Effect of Silk Protein on Ceramide Synthesis and the Expression of Ceramide Metabolic Enzymes in the Epidermis of NC/Nga Mice (실크단백질의 식이 공급이 아토피 피부염 동물 모델 NC/Nga Mice 피부의 세라마이드 함량 및 관련인자 발현에 미치는 영향)

  • Park, Kyung-Ho;Choi, Young-Sim;Kim, Hyun-Ae;Lee, Kwang-Gill;Yeo, Joo-Hong;Jung, Do-Hyun;Kim, Sung-Han;Cho, Yun-Hi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.5
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    • pp.554-562
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    • 2007
  • Ceramide rich intercellular lipid lamellae are thought to be particularly important in maintaining the structural integrity of epidermal barrier. Ceramide is synthesized de novo by serine palmitoyltransferase (SPT) phospholipid intermediates, serine and palmitic acid persist within the stratum corneum. The ceramide which is synthesized is degraded with fatty acid and sphingosine by degradative enzyme ceramidase. The depletion of ceramide in stratum corneum was reported in the atopic dermatitis. As an effort to search for the dietary source for improving the level of ceramide in epidermis, the dietary effects of various-typed silk protein were compared. Seventy male NC/Nga mice, an animal model of atopic dermatitis, were divided into seven groups: group CA as an atopic control with control diet, group S: 1% crude sericin diet, group F: 1% crude fibroin diet, group PS : peptide pattern of sericin(Mw 5000), group PF: peptide pattern of fibroin (Mw 1500), group AS: manufactured the same as amino acid profile of sericin and group AF: manufactured the same as amino acid profile of fibroin. Ten male BALB/c mice were served as group C (control group) control diet. All mice were fed on diet and water ad libitum for 10 weeks. Dry skin condition was established in group CA as ceramide content was decreased. Despite a marked decrease of mRNA and prorein expression of SPT, enzyme do novo synthesis, ceramide content of group S was dramatically increased by inhibiting the mRNA and protein expression of degradative enzyme ceramidase. However, dietary supplementation of crude silk fibroin protein (group F) and in other groups that were supplemented with either amino acid or peptide type of sericin or fibroin did not increase the level of ceramide. Together, our data demonstrate that dietary supplementation of crude sericin is more effective at improving ceramide level in epidemis of NC/Nga mice.

A Convenient Radiolabeling of [$^{11}$C](R)-PK11195 Using Loop Method in Automatic Synthesis Module ($^{11}$C 표지 자동합성장치에서 루프법을 이용한 ($^{11}$C)(R)-PK11195의 간편한 합성법)

  • Lee, Hak-Jeong;Jeong, Jae-Min;Lee, Yun-Sang;Kim, Hyung-Woo;Choi, Jae-Yeon;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.337-343
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    • 2009
  • Purpose: ((R)-1-(2-chlorophenyl)-N-1-[$^{11}$C]methyl-N(1-propyl)-3-isoquinoline carboxamide ((R)-PK11195) is a specific ligand for the peripheral type benzodiazepine receptor and a marker of activated microglia, used to measure inflammation in neurologic disorders. We report here that a direct and simple radiosynthesis of [$^{11}$C](R)-PK11195 in mild condition using NaH suspension in DMF and one-step loop method. Materials and Methods: (R)-N-Desmethyl-PK11195 (1 mg) in DMSO (0.1 mL) and NaH suspension in DMF (0.1 mL) were injected into a semi-prep HPLC loop. [$^{11}$C]methyl iodide was passed through HPLC loop at room temperature. Purification was performed using semi-preparative HPLC. Aliquots eluted at 11.3 min were collected and analyzed by analytical HPLC and mass spectrometer. Results: The labeling efficiency of [$^{11}$C](R)-PK11195 was 71.8$\pm$8.5%. The specific activity was 11.8:$\pm$6.4 GBq/$\mu$mol and radiochemical purity was higher than 99.2%. The mass spectrum of the product eluted at 11.3 min showed m/z peaks at 353.1 (M+1), indicating the mass and structure of (R)-PK11195. Conclusion: By the one-step loop method with the [$^{11}$C]CH3l automated synthesis module, [$^{11}C$](R)-PK11195 could be easily prepared in high radiochemical yield using NaH suspension in DMF.

Effect of Shading Levels on the Soil Properties, Growth Characteristics, and Chlorophyll Contents of Ligularia stenocephala (차광정도가 곤달비의 토양변화, 생육상황 및 엽록소 함량에 미치는 영향)

  • Park, Byoung-Mo;Kim, Chang-Hwan;Bae, Jong-Hyang;Shin, Jung-Ryeul
    • Journal of Bio-Environment Control
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    • v.20 no.4
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    • pp.352-356
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    • 2011
  • It is true that the industrial development has usually been accompanied with urbanization or centralization of population that has inevitably led to high-rise buildings and densely built-up living area in the cities. While it is badly needed to acquire as much green land within the city limits as possible to compensate for reduced space for recreational purpose in parallel with increasingly urbanized area, the living conditions of plants have become seriously devastated due to shortage of sun light walled-off by high-rise buildings and contaminated environment and air. The shade that is generated by high-rise and compact buildings hinders growth of plants, which makes it urgent to develop native ground cover plant that is strongly viable in the shade. For this purpose, Ligularia stenocephala, best known as greens for Ssam (rice and condiments wrapped in leaves) was cultivated under the 30%, 50%, and 80% shadings and observed to see if there would be any changes in soil conditions, growth of plants and chlorophyll contents depending on the shading rate. The leaf number was 10.8 pieces under the 50% shading and 8.4 under the 30%-shading, 7.7 pieces more than that cultivated under lighting. The leaf width turned out to be excellent from cultivation under the 50%- shading, an evidence indicating its possibility of being cultivated as native ground cover plant in the shade. The live weight of the plants cultivated under the shading increased to 31.63 g, 43.39 g and 19.40 g, respectively, compared to 90.43 g of those in the untreated control plot. The increase in growth of roots was particularly significant with 48.48 g in comparison to 12.33 g under 30% shading cultivation. The chlorophyll synthesis amounted to 46.2 under the 50% shading, showing an increase compared to 41.9 under lighting. The chlorophyll synthesis rather shrank under other shading conditions. The cultivation of Ligularia stenocephala under the 50% shading showed the best condition in growth as native ground cover plant.

Synthesis and Photovoltaic Properties of Low Band Gap π-Cojugated Polymer Based on 4,7-Di-thiophen-2-yl-benzo[1,2,5]thiadiazole (4,7-Di-thiophen-2-yl-benzo[1,2,5]thiadiazole을 기본으로 한 고분자의 합성 및 광전변환 특성)

  • Shin, Woong;You, Hyeri;Park, Jeong Bae;Park, Sang Jun;Jeong, Mi Seon;Moon, Myung-Jun;Kim, Joo Hyun
    • Applied Chemistry for Engineering
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    • v.21 no.2
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    • pp.137-141
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    • 2010
  • Poly [4,7-Di-thiophen-2-yl-benzo(1,2,5)thiadiazole]-alt-1,4-bis(dodecyloxy)-2,5-divinylbenzene (PPVTBT) was synthesized by the Heck coupling reaction between 4,7-Di-thiophen-2-yl-benzo(1,2,5)thiadiazole and 1,4-bis(dodecyloxy)-2,5-divinylbenzene. The maximum absorption and band gap of PPVTBT were 550 nm and 1.74 eV, respectively. The HOMO and LUMO energy level of PPVTBT were -5.24 eV and -3.50 eV, respectively. The photovoltaic device based on the blend of PPVTBT and (6)-1-(3-(methoxycarbonyl)propyl)-{5}-1-phenyl[5,6]-$C_{61}$ (PCBM) (1 : 6 by weight ratio) was fabricated. The efficiency of device was 0.16%. The short circuit current density (Jsc), fill factor (FF) and open-circuit voltage (Voc) of the device was $0.74mA/cm^{2}$, 31% and 0.71 V, respectively, under AM 1.5 G and 1 sun condition ($100mA/cm^{2}$).