청가시덩굴 추출물의 기능성 원료 표준화를 위한 지표성분 Resveratrol, trans-Scirpusin A의 분석법 개발 및 검증

권진관(Jin Gwan Kwon),정연우(Yeon Woo Jung),최윤혁(Yun-Hyeok Choi),이지은(Ji Eun Lee),정원식(Wonsik Jeong),이정아(Jung A Lee),최춘환(Chun Whan Choi),안은경(Eun-Kyung Ahn),최용문(Yongmun Choi),홍성수(Seong Su Hong) 한국식품영양과학회 2022 한국식품영양과학회지 Vol.51 No.11

본 연구는 HPLC를 이용하여 청가시덩굴 추출물을 개별인정형 건강기능식품의 기능성 원료로 개발하기 위한 원료 표준화의 일환으로, 청가시덩굴 추출물의 지표성분을 resveratrol과 trans-scirpusin A로 설정하고 이에 대한 HPLC 분석법을 확립하여 유효성의 검증을 실시하였다. 분석법 유효성 검증은 특이성, 직선성, 정확도, 정밀도, 검출한계 및 정량한계 등을 통해 분석법의 신뢰성을 검증하였으며, 그 결과 표준용액과 청가시덩굴 추출물 간의 HPLC 크로마토그램 및 UV spectrum의 일치 여부 등의 비교를 통해 다른 물질과 간섭 없이 피크가 분리된 것으로 특이성을 확인하였다. 또한 표준용액 검량선의 상관계수(R²)는 0.9999로 매우 우수한 직선성으로 관찰되어 분석에 적합한 것으로 확인되었으며, 검량선의 기울기 및 표준편차를 이용한 검출한계는 resveratrol이 0.98 μg/mL, trans-scirpusin A는 0.49 μg/mL였고 정량한계는 resveratrol이 2.98 μg/mL, trans-scirpusin A는 1.48 μg/mL로 각각 확인되었다. 청가시덩굴 추출물에 표준물질을 3개 농도 첨가하고 분석한 회수율은 resveratrol이 98.77~99.24%, trans-scirpusin A는 98.45~99.45%로 나타나 정확성이 있는 것을 확인할 수 있었다. 청가시덩굴 추출물의 조제 농도 2.2, 4.4 및 6.6 mg/mL에서 반복성은 resveratrol이 0.99~1.22%, trans-scirpusin A는 1.12~1.32%를, 실험실 내 정밀성에서는 일내 정밀성은 resveratrol이 0.67~0.87%, trans-scirpusin A는 1.18~1.33%로 나타났고 일간 정밀성은 resveratrol이 0.93~1.22%, trans-scirpusin A는 1.33~2.27%로 확인되어 본 분석법은 정밀성이 있음을 확인할 수 있었다. 이상의 분석결과를 통해 확립된 청가시덩굴 추출물의 지표성분인 resveratrol과 trans-scirpusin A의 HPLC 분석법은 적합한 시험법으로 검증되었으며, 본 시험법은 향후 청가시덩굴 추출물의 건강기능식품 기능성 원료 개발과 표준화를 위한 기초자료로 활용될 것으로 사료된다. This study was undertaken to establish an analytical method using high-performance liquid chromatography (HPLC). HPLC for the standard determination of resveratrol and trans-scirpusin A as functional ingredients in Smilax sieboldii extract. We evaluated the specificity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ) of various analytical methods for detecting resveratrol and trans-scirpusin A using HPLC. The specificity was confirmed by the chromatogram obtained using the HPLC analytical method. Also, the results of UV and the coefficient of correlation (R²) obtained was 0.999, which confirmed that this was a suitable analysis with high linearity. The LOD was 0.98, 0.49 μg/mL, and LOQ was 2.98, 1.48 μg/mL, which was confirmed as a suitable limit level for the analysis of resveratrol and trans-scirpusin A content in the S. sieboldii extract. The recovery of resveratrol and trans-scirpusin A content was determined to be 98.77±0.73∼99.24±1.47% and 98.45±1.18∼99.45±1.66%, respectively, indicating high accuracy. The intra-day repeatability and the intra-laboratory precision of the daily repetition were confirmed to be 0.67∼0.87%, 1.18∼1.33% and 0.93∼1.22%, 1.33∼2.27%, respectively, for trans-scirpusin A, for the relative standard deviation. These results indicate that the reported HPLC method is simple, reliable, and reproducible for the detection of resveratrol and trans-scirpusin A in S. sieboldii extract.

Studies on Simultaneous Determination of Chlorophyll a and b, Pheophorbide a, and $\beta-Carotene$ in Chlorella and Spirulina Products

이영자,김소희,김진숙,한정아,서해점,임효정,최수영,Lee Young Ja,Kim So Hee,Kim Jin-Sook,Han Jeong A,Seo Hae Jeom,Lim Hyo Jeong,Choi Soo Young The Korean Society of Food Hygiene and Safety 2005 한국식품위생안전성학회지 Vol.20 No.3

역상 컬럼을 이용하여 건강기능식품 중 클로렐라 및 스피루리나제품에 함유되어 있는 엽록소 a, b,페오포르바이드 a 및 $\beta$-카로틴의 HPLC동시분석법을 확립하였으며, 첨가농도 $50\;\mug/ml$에서 엽록소 a, b, 페오포르바이드 a 및 $\beta$-카로틴에 대한 회수율시험결과, 각각 2.8, 6.0, 10.6 및 $10.4\%$의 상대표준편차와 70.3, 71.6, 60.1 및 $90.5\%$의 회수율을 각각 나타냈다. 이때 검출한계는 $0.1\sim1.0\;\mug/ml$, 정량한계는 $0.2\sim2.0\;\mug/ml$이었으며 검량선 상관계수도 0.995 이상의 직선성을 보여주었다. 국내유통 클로렐라 및 스피루리나제품에 대한 엽록소 a, b,페오포르바이드 a및 $\beta$-카로틴의 함유량을 분석한 결과 엽록소 a $121.g\sim543$, 엽록소 b $0.6\sim160.0$, 페오포르바이드 a 및 P-카로틴 $383.6\sim1713.7mg/ml$ 수준으로 나타났다. 엽록소 b의 함유량은 클로렐라제품에서 평균 374.0 mg/100 g 으로 스피루리나제품의 평균 10.5 mg/100 g 보다 30배 이상 함유하고 있는 것으로 확인되었다. 그러나 $\beta$-카로틴의 함유량은 스피루리나제품이 평균 1335.4 mg/100 g 로 클로렐라제품의 평균 495.0 mg/100 g 보다 평균 함유량에서 2.7배 높은 것으로 나타났다. 국내 건강기능식품공전 중 클로렐라 및 스피루리나제품의 엽록소 a b, 및 페오포르바이드 항목의 규격검사를 본 연구의 동시분석법으로 개정함으로써 각 성분 함량의 정량, 분석시간의 단축 및 비용절감 둥 시험방법을 크게 개선할 수 있을 것으로 기대된다. A simple and sensitive analysis method based on reverse phase (RP) HPLC with UV detector was developed for simultaneous determination of chlorophyll a and b, pheophorbide a and $\beta-Carotene$ in Chlorella and Spirulina products. For added concentration $(50\;\mug/ml)$ of chlorophyll a and b, pheophorbide a and $\beta-Carotene$, recoveries of those were 70.3, 71.6, 60.1 and $90.5\%$, respectively, with relative standard deviations of 2.8,6.0, 10.6 and $10.4\%$. Limit of detection and quantification had ranges of $0.1\sim1.0\;\mug/ml$ and $0.2\sim2.0\;\mug/ml$, respectively. Calibration curve was linear with correlation coefficient of 0.995 for chlorophyll a and b, pheophorbide a and $\beta-Carotene$. Results of simultaneous determination in Chlorella and Spirulina products were showed ranges of $121.g\sim543.0\;\mug/ml$ for chlorophyll a,$0.6\sim160.0\;\mug/ml$ for chlorophyll b, $19.2\sim60.3\;\mug/ml$ for pheophorbide a and $383.6\sim1713.7\;\mug/ml$ for $\beta-Carotene$, respectively. Chlorophyll b contents in Chlorella products were detected above 30 times level to those in Spirulina products. $\beta-Carotene$ contents in Spirulina products were detected 2.7 times level to those in Chlorella products.

Polypharmacology of <i>N</i>⁶-(3-Iodobenzyl)adenosine-5′-<i>N</i>-methyluronamide (IB-MECA) and Related A₃ Adenosine Receptor Ligands: Peroxisome Proliferator Activated Receptor (PPAR) γ Partial Agonist and PPARδ Antagonist Activ

Yu, Jinha,Ahn, Seyeon,Kim, Hee Jin,Lee, Moonyoung,Ahn, Sungjin,Kim, Jungmin,Jin, Sun Hee,Lee, Eunyoung,Kim, Gyudong,Cheong, Jae Hoon,Jacobson, Kenneth A.,Jeong, Lak Shin,Noh, Minsoo American Chemical Society 2017 Journal of medicinal chemistry Vol.60 No.17

<P>A(3) adenosine receptor (AR) ligands including A(3) AR agonist, N-6-(3-iodobenzyl)adenosine-5'-N-methyluronamide (1a, IB-MEGA) were examined for adiponectin production in human bone marrow mesenchymal stem cells (hBM-MSCs). In this model, 1a significantly increased adiponectin production, which is associated with improved insulin sensitivity. However, A(3) AR antagonists also promoted adiponectin production in hBM-MSCs, indicating that the A(3) AR pathway may not be directly involved in the adiponectin promoting activity. In a target deconvolution study, their adiponectin-promoting activity was significantly correlated to their binding activity to both peroxisome proliferator activated receptor (PPAR) gamma and PPAR delta. They functioned as both PPAR gamma partial agonists and PPAR delta antagonists. In the diabetic mouse model, la and its structural analogues A(3) AR antagonists significantly decreased the serum levels of glucose and triglyceride, supporting their antidiabetic potential. These findings indicate that the polypharmacophore of these compounds may provide therapeutic insight into their multipotent efficacy against various human diseases.</P>

귀충만감 환자에 대한 임상적 고찰

이상철,정진호,이상목,이재완,엄재욱,박춘근 釜算慶南耳鼻咽喉科學會 1994 임상이비인후과 Vol.5 No.1

Polyphenolic compounds from Korean <i>Lonicera japonica</i> Thunb. induces apoptosis via AKT and caspase cascade activation in A549 cells

Park, Kwang Il,Park, Hyeonsoo,Nagappan, Arulkumar,Hong, Gyeong Eun,Yumnam, Silvia,Lee, Ho Jeong,Kim, Eun Hee,Lee, Won Sup,Shin, Sung Chul,Kim, Jin A,Lee, Sang Joon,Ma, Jin Yeul,Min, Taesun,Heo, Jeong D.A. Spandidos 2017 Oncology letters Vol.13 No.4

<P><I>Lonicera japonica</I> Thunb. (<I>L. japonica</I> T.) has historically been used in Korean herbal medicine due to its anticancer and protective effects on the respiratory system. In the present study, the polyphenolic compounds in <I>L. japonica</I> T. were investigated using high-performance liquid chromatography coupled with tandem mass spectrometry, and its anticancer effects on A549 non-small-cell lung cancer cells were studied. Polyphenolic compounds potentially inhibit A549 cells in a dose-dependent manner. Flow cytometry and western blot analysis demonstrated that polyphenolic compounds induce apoptosis by regulating the protein expression levels of caspases, poly-(ADP-ribose) polymerase and the B-cell lymphoma-2-associated X-protein/B-cell lymphoma-extra large ratio. Furthermore, polyphenolic compounds inhibited mitochondrial membrane potential activity. Caspase-3 activity was increased in a dose-dependent manner and polyphenolic compounds inhibited the activation of protein kinase B by dephosphorylation. These results suggest that polyphenolic compounds in A549 cells indicate the anticancer activity through the induction of apoptosis.</P>

Human coagulation factor VIII domain-specific recombinant polypeptide expression

Su Jin Choi,Ki Jung Jang,Jeong-A Lim,Hye Sun Kim 대한혈액학회 2015 Blood Research Vol.50 No.2

BackgroundHemophilia A is caused by heterogeneous mutations in F8. Coagulation factor VIII (FVIII),the product of F8, is composed of multiple domains designated A1-A2-B-A3-C1-C2. FVIIIis known to interact with diverse proteins, and this characteristic may be important forhemostasis. However, little is known about domain-specific functions or their specificbinding partners. MethodsTo determine F8 domain-specific functions during blood coagulation, the FVIII domainsA1, A2, A3, and C were cloned from Hep3B hepatocytes. Domain-specific recombinantpolypeptides were glutathione S-transferase (GST)- or polyhistidine (His)-tagged,over-expressed in bacteria, and purified by specific affinity chromatography. ResultsRecombinant polypeptides of predicted sizes were obtained. The GST-tagged A2 polypeptideinteracted with coagulation factor IX, which is known to bind the A2 domain of activatedFVIII. ConclusionRecombinant, domain-specific polypeptides are useful tools to study the domain-specificfunctions of FVIII during the coagulation process, and they may be used for productionof domain-specific antibodies.

한국인 직무 스트레스 측정도구의 개발 및 표준화

장세진,고상백,강동묵,김성아,강명근,이철갑,정진주,조정진,손미아,채창호,김정원,김정일,김형수,노상철,박재범,우종민,김수영,김정연,하미나,박정선,이경용,김형렬,공정옥,김인아,김정수,박준호,현숙정,손동국 大韓産業醫學會 2005 대한직업환경의학회지 Vol.17 No.4

Background and Purposes: Over the past three decades, numerous studies performed in Korea have reported that job stress is a determinant risk factor for chronic diseases and work disability. Every society has its own culture and occupational climate particular to their organizations, and hence experiences different occupational stress. An occupational stress measurement tool therefore needs to be developed to estimate it objectively. The purpose of this study is to develop and standardize the Korean Occupational Stress Scale (KOSS) which is considered to be unique and specific occupational stressors in Korean employees. Subjects and Methods: Data were obtained from the National Study for Development and Standardization of Occupational Stress (NSDSOS Project: 2002-2004). A total of 12,631 employees from a nationwide sample proportional to the Korean Standard Industrial Classification and the Korean Standard Occupational Classification were administered. The KOSS was developed for 2 years (2002-2004). In the first year, we collected 255 items from the most popular job stress measurement tools such as JCQ, ERI, NIOSH and OSI, and 44 items derived from the a qualitative study (depth interview). Forty-three items of KOSS, in the second year, were retained for use in the final version of the KOSS by using Delphi and factor analysis. Items were scored using conventional 1-2-3-4 Likert scores for the response categories. Results: We developed eight subscales by using factor analysis and validation process: physical environment (3 items), job demand (8 items), insufficient job control (5 items), interpersonal conflict (4 items), job insecurity (6 items), organizational system (7 items), lack of reward (6 items), and occupational climate (4 items). Together they explained 50.0% of total variance. Internal consistency alpha scores were ranged from 0.51 to 0.82. Twenty-four items of the short form of the KOSS (KOSS-SF) were also developed to estimate job stress in the work setting. Because the levels of the subscales of occupational stress were gender dependent, gender-specific standard norms for both the 43-item full version and the 24-item short form using a quartile for the subscales of KOSS were presented. Conclusion: The results of this study suggest that KOSS might be an appropriate measurement scale to estimate occupational stress of Korean employees. Further and more detailed study needs to be conducted to improve the validity of this scale.

Human coagulation factor VIII domain-specific recombinant polypeptide expression

Choi, Su Jin,Jang, Ki Jung,Lim, Jeong-A,Kim, Hye Sun Korean Society of Hematology; Korean Society of Bl 2015 Blood Research Vol.50 No.2

<P><B>Background</B></P><P>Hemophilia A is caused by heterogeneous mutations in <I>F8</I>. Coagulation factor VIII (FVIII), the product of <I>F8</I>, is composed of multiple domains designated A1-A2-B-A3-C1-C2. FVIII is known to interact with diverse proteins, and this characteristic may be important for hemostasis. However, little is known about domain-specific functions or their specific binding partners.</P><P><B>Methods</B></P><P>To determine <I>F8</I> domain-specific functions during blood coagulation, the FVIII domains A1, A2, A3, and C were cloned from Hep3B hepatocytes. Domain-specific recombinant polypeptides were glutathione S-transferase (GST)- or polyhistidine (His)-tagged, over-expressed in bacteria, and purified by specific affinity chromatography.</P><P><B>Results</B></P><P>Recombinant polypeptides of predicted sizes were obtained. The GST-tagged A2 polypeptide interacted with coagulation factor IX, which is known to bind the A2 domain of activated FVIII.</P><P><B>Conclusion</B></P><P>Recombinant, domain-specific polypeptides are useful tools to study the domain-specific functions of FVIII during the coagulation process, and they may be used for production of domain-specific antibodies.</P>

협응이동훈련이 아동의 자세 불균형과 보행에 미치는 영향 : 단일사례설계

이정아 ( Jeong-a Lee ),김진철 ( Jin-cheol Kim ) 대한물리의학회 2019 대한물리의학회지 Vol.14 No.3

PURPOSE: This study was examined the effects of coordinative locomotor training (CLT) on the postural imbalance and gait in children. METHODS: Four children were sampled as subjects. A single subject study (A-B-A’) was conducted by measuring the following: baseline five sessions;, intervention phase, 15 sessions;, and postline (A’) five sessions. The research period was eight weeks. The CLT program consisted of warming-up exercise, main exercise, and finishing exercise, and it was performed for one hour per day. A oneleg standing test (OLST) was performed determine the static balance. A functional reach test (FRT) was performed determine the reactionary balance. To determine the dynamic balance, the time up and go test (TUG) was performed. A 10m walking test (10 MWT) was performed to determine the walking ability. A statistical test was performed through descriptive statistics to present the average and standard deviation, and the variation rate was compared using a visual analysis method with graphs. RESULTS: As a result of CLT application, all four subjects improved the OLST, FRT, TUG, and 10 MWT compared to the intervention period baseline, and postline period. CONCLUSION: CLT appeared to improve the posture imbalance and gait in children.

Clinical Impact of Exosomal microRNA as a Novel Biomarker of Liver Fibrosis

( Young Chang ),( Jae-a Han ),( Suk Min Kang ),( Soung Won Jeong ),( Tom Ryu ),( Han Seul Park ),( Jeong-ju Yoo ),( Sae Hwan Lee ),( Sang Gyune Kim ),( Young Seok Kim ),( Hong Soo Kim ),( So Young Jin 대한간학회 2020 춘·추계 학술대회 (KASL) Vol.2020 No.1

Aims: Many approaches have been suggested for the non-invasive diagnosis of liver fibrosis, including the use of serum biomarkers and ultrasound-based elastography, but none has yet replaced liver biopsy. MicroRNAs (miRNAs) have been suggested as potential diagnostic tools for liver diseases. We investigated alterations in the expression of serum exosomal miRNAs with the progression of liver fibrosis and evaluated their clinical applicability as biomarkers. Methods: This study prospectively enrolled 71 patients who underwent liver biopsy at a large-volume academic hospital in Korea. Exosomes were extracted from serum samples, and next-generation sequencing (NGS) of miRNAs was conducted in patients from different stages of liver fibrosis. Differential expression of miRNAs was quantified using targeted real-time quantitative polymerase chain reaction (RT-qPCR). A model was derived to discriminate advanced fibrosis based on miRNA levels using multivariate logistic regression. The performance of this model was evaluated and compared using area under the receiver operator characteristic (ROC) curve (AUC) and De- Long’s test. Results: NGS data revealed the relationship between exosomal miR-122 expression and liver fibrosis progression. The level of miR-122 decreased as the pathologic fibrosis grade progressed from stage 0 to 4. Patients with biopsy-proven advanced fibrosis had significantly lower levels of exosomal miR-122 (P<0.001) than those without advanced fibrosis. Exosomal miR-122 exhibited a fair performance in discriminating advanced fibrosis with an AUC of 0.77, which improved to 0.86 in combination with fibrosis-4 score (FIB-4) and transient elastography (TE). This value was higher than that reported for any other non-invasive modalities, including TE (AUC of 0.80) or FIB-4 (AUC of 0.57) alone. In a subgroup of patients with a non-viral etiology of liver disease, the performance of exosomal miR-122 as a biomarker improved, evident from the increase in the AUC value to 0.87. In this subpopulation, the combination model of miR- 122, FIB-4, and TE showed the best discrimination ability (AUC of 0.90), which was significantly higher than that of TE alone (AUC of 0.83; DeLong’s test P=0.046). Inhibition of miR-122 expression increased the proliferation of the human hepatic stellate cell line, LX-2, and upregulated the expression of collagen- 1A, a-smooth muscle actin, fibronectin, and transforming growth factor-ß. Conclusions: Exosomal miR-122 may serve as a novel biomarker for discriminating advanced liver fibrosis, and its accuracy may enhanced in combination with other non-invasive tests such as FIB-4 and TE.