LC-MS/MS를 이용한 채소류 및 서류의 클로로젠산 함량 조사

류지은(Ji-Eun Ryu),김운호(Woon-Ho Kim),배호정(Ho-Jeong Bae),정유정(You-Jung Jung),최영주(Young-Ju Choi),문경은(Kyeong-Eun Moon),최종철(Jong-Chul Choi),채경석(Kyung-Suk Chae),이진희(Jin-Hee Lee),도영숙(Young-Sook Do),최옥경(Ok-Kyung C 한국식품영양과학회 2020 한국식품영양과학회지 Vol.49 No.8

본 연구는 LC-MS/MS를 이용해 다소비 채소류 및 서류에 함유된 9종의 chlorogenic acid 이성질체를 분석하기 위하여 시료 추출 후 QuEChERS kit으로 정제하는 전처리법을 확립하였고, Syncronis C18 컬럼을 사용하여 30분 동안 분석하는 이동상 용매 조성과 9종의 이성질체의 정량 및 정성 이온을 선택하여 negative 모드에서 분석하는 기기 분석법을 확립하였다. 확립한 분석법은 AOAC 가이드라인에 따라 특이성, 직선성, 검출한계, 정량한계, 정확성, 정밀성을 확인하여 유효성을 검증하였다. 다소비 채소류 및 서류 43품목(48품종)의 chlorogenic acids 함량을 분석한 결과, 24품목(29품종)에 함유되었음을 확인하였다. 채소류의 경우 39품목(42품종) 중 22품목(25품종)에 chlorogenic acids가 함유되었으며(0.01±0.01~38.31±8.73 ㎎/100 g FW), 취나물, 케일, 미나리, 우엉, 상추, 브로콜리, 콜리플라워, 가지, 치커리, 도라지 순으로 함량이 높게 나타났다. 서류는 고구마(호박고구마, 밤고구마), 감자(로즈밸리, 수미), 마, 토란 등 4품목(6품종)을 분석하였는데, 고구마의 경우 밤고구마(1.31±0.52 ㎎/100 g FW)보다 호박고구마(1.42±0.39 ㎎/100 g FW)에, 감자의 경우 수미(0.09±0.03 ㎎/100 g FW)보다 로즈밸리(0.37±0.07 ㎎/100 g FW)에 chlorogenic acids 함량이 높게 나타났다. 또한 취나물, 미나리, 고구마, 감자를 대상으로 조리시간에 따른 chlorogenic acids 함량 변화를 확인한 결과, 취나물과 미나리는 30초 삶았을 때 가장 높은 함량을 나타내었고, 감자는 30분, 고구마는 20분쪘을 때 chlorogenic acids 함량이 높게 나타났다. 본 연구에서는 다소비 채소류 및 서류의 chlorogenic acids 함량과 조리시간에 따른 chlorogenic acids 함량 변화를 확인하여 식품 내 chlorogenic acids 함량에 관한 기초자료로 제공하고자 하였다. In this study, a sample preparation method and simultaneous determination method were developed to determine the chlorogenic acids content of frequently consumed vegetables, root and tuber crops in Korea by liquid chromatography coupled to mass spectrometry. The developed method was validated for its specificity, linearity, limit of detection, limit of quantification, recovery, accuracy, and precision based on the AOAC guidelines. Among the 39 items of vegetables, 22 items (0.01±0.01∼38.31±8.73 ㎎/100 g fresh weight (FW)) were quantified. The chlorogenic acids content of the vegetables was in the following order: chwinamul (38.31±8.73 ㎎/100 g FW), kale (22.93±0.75 ㎎/100 g FW), water dropwort (10.67±4.46 ㎎/100 g FW), burdock root (9.72±2.52 ㎎/100 g FW), red lettuce (leaf) (8.66±5.37 ㎎/100 g FW), broccoli (2.45±0.50 ㎎/100 g FW), and cauliflower (2.35±0.36 ㎎/100 g FW). Among the four items of root and tuber crops, sweet potato (1.42±0.39 ㎎/100 g FW) and potato (0.37±0.07 ㎎/100 g FW) were quantified. In addition, the chlorogenic acids content of chwinamul and water dropwort was highest when boiled for 30 seconds. Sweet potato steamed for 20 minutes and potato steamed for 30 minutes showed high chlorogenic acids contents.

반응표면분석법을 이용한 구절초의 Linarin, Chlorogenic Acid 추출 조건 최적화

최대호,정은숙,강정목,박용우 경상대학교 농업생명과학연구원 2024 농업생명과학연구 Vol.58 No.4

구절초(Chrysanthemum zawadskii var. latilobum (Maxim.) Kitam.)는 linarin, chlorogenic acid 등의 유용성분들을 함유하고 있으며, 산업적으 로 이용가치가 있는 식물이다. 구절초의 산업화를 위해 유용성분인 linarin과 chlorogenic acid의 최적 추출 조건을 구하기 위해 반응표면분석을 수행하였으며, Linarin의 최적 추출 조건은 에탄올 농도 100%, 추출 시간 30h, 추출 온도 75℃였다. 최적 추출 조건에서 예상된 linarin의 값은 33.36 mg/g였으며 실험값은 38.33±2.66 mg/g였다. Chlorogenic acid의 최적 추출 조건은 에탄올 농도 48%, 추출 시간 30h, 추출 온도 65℃로 예상된 chlorogenic acid의 함량은 42.19 mg/g였으며 실험값은 44.34±2.09 mg/g이었다. 반응표면분석법으로 도출된 최적 추출 조건을 통해 구절초에서 linarin과 chlorogenic acid의 효율적인 추출이 가능함을 확인하였다. Chrysanthemum zawadskii var. latilobum contains useful components such as linarin and chlorogenic acid, and is a plant with industrial value. For the industrialization of C. zawadskii var. latilobum, response surface analysis was performed to determine the optimal extraction conditions for the useful components linarin and chlorogenic acid. The optimal extraction conditions for linarin were 100% ethanol concentration, 30h extraction time, and 75℃ extraction temperature. The expected value of linarin under optimal extraction conditions was 33.36 mg/g, and the experimental value was 38.33±2.66 mg/g. The optimal extraction conditions for chlorogenic acid were 48% ethanol concentration, 30h extraction time, and 65℃ extraction temperature. The expected chlorogenic acid content was 42.19 mg/g, and the experimental value was 44.34±2.09 mg/g. It was confirmed that efficient acquisition of linarin and chlorogenic acid was possible through optimal extraction conditions derived by response surface analysis.

블루베리 효모 발효 분말의 기능성 원료 표준화를 위한 지표성분으로서 Chlorogenic Acid 분석 및 검증

정수지(Su-Ji Jeong),양희종(Hee-Jong Yang),류명선(Myeong Seon Ryu),서지원(Ji Won Seo),정도연(Do-Youn Jeong) 한국식품영양과학회 2019 한국식품영양과학회지 Vol.48 No.10

블루베리 효모 발효 분말의 개별 인정형 건강기능식품 기능성 원료로 개발하기 위하여 지표성분 표준화를 위한 chlorogenic acid의 분석법 설정과 분석법에 대한 벨리데이션을 실시하였다. 벨리데이션에 앞서 chlorogenic acid의 분석을 위한 추출 용매의 농도별 수율을 조사한 결과 50% 에탄올로 추출 시 2,040.14 μg/mL로 가장 우수한 수율을 나타내어 추출 용매의 농도를 확립하였다. Chlorogenic acid의 분석은 1% acetic acid가 첨가된 water와 acetonitrile을 이동상으로 하고 Capcellpak-C18(4.6×150 mm, 5 μm) 컬럼을 사용하여 기울기 용리(gradient elution) 방법으로 분석하였다. 본 연구에서는 분석법에 대한 특이성, 직선성, 정확성과 정밀성 및 회수율을 확인하였다. Chlorogenic acid의 검량선은 상관계수(R<SUP>2</SUP>)가 0.9999로 우수한 직선성을 보였으며, 검출한계와 정량한계는 각각 5.38 μg/mL, 16.32 μg/mL로 나타났다. 시료 농도별 intra-day 및 inter-day 분석에서의 상대표준편차(RSD)는 각각 1.34~2.5%, 0.92~2.17%로 나왔다. 회수율 측정 결과에서는 103.03~103.36%로 나왔으며, RSD는 0.83~1.22%였다. 따라서 HPLC를 이용한 chlorogenic acid의 분석법이 블루베리 효모 발효 분말의 기능성 원료 표준화를 위한 지표성분 분석의 적합한 시험법임을 검증하였다. 또한 본 시험법에 따라 제조 공정별 블루베리 효모 발효 분말 내의 chlorogenic acid 함량을 측정하였으며, 최종 산물인 블루베리 효모 발효 분말의 chlorogenic acid 함량을 세 lot를 3회 분석하였을 때 2,032.66 μg/mL가 나왔으며, RSD 값은 0.58%로 확인되었다. 따라서 본 연구를 통해 확립한 분석법이 블루베리 효모 발효 분말의 개별인 정형 건강기능식품 기능성 원료 개발을 위한 기초 자료로 활용될 것으로 사료된다. This study validated an analytical method for chlorogenic acid in blueberry yeast fermented powder (BYFP) for standardization as a functional health ingredient. Measurements of the yield according to the concentration of the extraction solvent for analyzing the level chlorogenic acid before validation showed that the extraction yield was highest at 2,040.14 μg/mL when extracted with 50% ethanol. The quantitative determination method of chlorogenic acid was optimized by HPLC analysis using a Capcellpak-C18 column with a gradient elution of 1% acetic acid in water and acetonitrile. This method was validated according to the specificity, linearity, accuracy, precision test, and recovery test. The method showed reliable linearity of the calibration curve with a R2 of 0.9999, and the limit of quantitation and the detection limit were 5.38 μg/mL and 16.32 μg/mL, respectively. The relative standard deviation (RSD) from the intra- and inter-day precision according to the chlorogenic acid concentration ranged from 1.34 to 2.5% and 0.92 to 2.17%, respectively. The recovery test ranged from 103.03% to 103.36% with RSD values from 0.83 to 1.22%. Therefore, chlorogenic acid as a marker compound in BYFP was analyzed. The amount of chlorogenic acid in BYFP was approximately 2,032.66 μg/mL after triplicate analysis, and the RSD was 0.58% using the validated method. These results suggest that the method is very useful for evaluating the marker compounds in BYFP to develop a health functional food material.

Chlorogenic acid alleviates the reduction of Akt and Bad phosphorylation and of phospho-Bad and 14-3-3 binding in an animal model of stroke

Murad-Ali Shah,Ju-Bin Kang,Myeong Ok Kim,Phil-Ok Koh 대한수의학회 2022 Journal of Veterinary Science Vol.23 No.6

Background: Stroke is caused by disruption of blood supply and results in permanent disabilities as well as death. Chlorogenic acid is a phenolic compound found in various fruits and coffee and exerts antioxidant, anti-inflammatory, and anti-apoptotic effects. Objectives: The purpose of this study was to investigate whether chlorogenic acid regulates the PI3K-Akt-Bad signaling pathway in middle cerebral artery occlusion (MCAO)-induced damage. Methods: Chlorogenic acid (30 mg/kg) or vehicle was administered peritoneally to adult male rats 2 h after MCAO surgery, and animals were sacrificed 24 h after MCAO surgery. Neurobehavioral tests were performed, and brain tissues were isolated. The cerebral cortex was collected for Western blot and immunoprecipitation analyses. Results: MCAO damage caused severe neurobehavioral disorders and chlorogenic acid improved the neurological disorders. Chlorogenic acid alleviated the MCAO-induced histopathological changes and decreased the number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells. Furthermore, MCAO-induced damage reduced the expression of phospho-PDK1, phospho-Akt, and phospho-Bad, which was alleviated with administration of chlorogenic acid. The interaction between phospho-Bad and 14-3-3 levels was reduced in MCAO animals, which was attenuated by chlorogenic acid treatment. In addition, chlorogenic acid alleviated the increase of cytochrome c and caspase-3 expression caused by MCAO damage. Conclusions: The results of the present study showed that chlorogenic acid activates phospho-Akt and phospho-Bad and promotes the interaction between phospho-Bad and 14-3-3 during MCAO damage. In conclusion, chlorogenic acid exerts neuroprotective effects by activating the Akt-Bad signaling pathway and maintaining the interaction between phospho-Bad and 14-3-3 in ischemic stroke model.

산초나무(Zanthoxylum schinifolium) 잎의 Chlorogenic Acid와 Hesperidin 정량분석을 위한 HPLC-DAD 분석법 개발 및 평가

강주영,노혜령,김봉규 경상대학교 농업생명과학연구원 2024 농업생명과학연구 Vol.58 No.4

본 연구는 Diode Array Detector (DAD)가 장착된 고성능 액체 크로마토그래피(HPLC)를 이용하여 산초나무 잎에 함유된 chlorogenic acid와 hesperidin을 동시에 분석할 수 있는 표준 정량법 개발을 확립하고자 실시하였다. 산초나무 잎의 chlorogenic acid와 hesperidin은 MS/MS 분석 및 표준물질과의 유지시간(RT), UV 흡광도 비교를 통해 확인하였고, 산초나무 잎의 지표물질로 선정하였다. 개발한 분석방법은 지표물질을 대상으로 특이성, 정확성, 정밀도, 검출한계, 정량한계, 선형성에 대해 최적화하고 검증하였다. HPLC 분석으로 두 화합물에 대한 검량선은 각각 0.99% 이상의 높은 상관계수(r2)를 보였다. Chlorogenic acid의 전체 회수율은 106.82%이었고, hesperidin의 전체 회수율은 107.37%였다. Chlorogenic acid의 검출한계(LOD)는 0.271μg/ml, Hesperidin의 검출한계(LOD)는 0.005μg/ml였다. Chlorogenic acid의 일간정밀도는 0.102% (RSD%), Hesperidin의 일간정밀도는 1.038% (RSD%)였다. 본 연구에서 개발한 HPLC-DAD 분석방법은 기능성 원료 인정을 위한 분석기준을 충족하였다. 따라서 본 연구에서 확립한 HPLC 분석법은 향후 산초나무 잎에서 유래한 생리활성물질의 대량추출 및 관련제품을 개발할 때 원료 표준화에 적용가능할 것이다. We attempted to establish a High-performance liquid chromatography (HPLC) analysis method with a Diode Array Detector (DAD) for standard quantification of chlorogenic acid and hesperidin, which are healthy functional food ingredients in leaves of Zanthoxylum schinifolium. The chlorogenic acid and hesperidin from Z. schinifolium leaves were identified by MS/MS analysis, comparing retention time and UV absorbance with authentic compounds. HPLC method was validated by specificity, accuracy, precision, and limit of quantification. The HPLC method showed that calibration curves for each compounds have high linearity with a correlation coefficient (r2) of > 0.99%. The total recovery rate of chlorogenic acid and hesperidin was 106.82% and 107.37%, respectively. The detection limits (LOD) for chlorogenic acid and hesperidin are 0.271㎍/ml and 0.005㎍/ml, respectively. Daily precision of chlorogenic acid was 0.102% (RSD%), and hesperidin was 1.038% (RSD%). All tested parameters met the appropriate ranges designated in the guideline. Therefore, the HPLC analysis method established in this study can be applied and used to standardize health-related functional products or pharmaceutical raw materials and products derived from Z. schinifolium leaves.

용매 분획 및 유산균 발효가 커피체리 펄프 페놀성 화합물에 미치는 영향

온지헌,오병민,오현화,정도연,김영수 한국식품영양과학회 2023 한국식품영양과학회지 Vol.52 No.3

This study was conducted to investigate the changes in the levels of phenolic compound of coffee cherry pulp (CCP) using solvent fractionation and lactic acid bacteria fermentation. Ethanol extracts of CCP were obtained using various ethanol ratios to evaluate their antioxidant contents and activities. The ethanol extract with the highest total phenolic contents (TPC) and total flavonoid contents (TFC) was fractionated according to the difference in the polarity of the solvent. TPC and TFC in the 80% ethanol extract were 46.55 mg gallic acid equivalent (GAE/g) and 113.42 mg quercetin equivalent (QE/g), respectively. The CCP extract with 80% ethanol solvent (CEE) was fractionated in order n-hexane, chloroform, ethyl acetate, and n-butanol. TPC and TFC in the ethyl acetate fraction (CEF) were 627.50 mg GAE/g and 1,632.31 mg QE/g, respectively. The IC50 value for DPPH and ABTS radical scavenging activity in CEF was evaluated at 0.17 mg/mL and 0.34 mg/mL, respectively. As a result of the comparison with the phe nolic compounds between CEE and CEF, 3-chlorogenic acid and 3,5-dicaffeoylquinic acid showed the highest increase with 15.05 to 62.75 mg/g and 20.10 to 72.26 mg/g, respectively. The caffeic acid contents of CMB, which was MRS broth contained CCP, fermented by Lactobacillus rhamnosus GG for 10 days was 1.51 μg/mL, which was more than two times that of unfermented CMB (0.75 μg/mL). Based on these results, we propose the use of lactic acid bacteria to increase the yield of caffeic acid from CCP. 본 연구에서는 커피체리 펄프에 함유된 항산화 성분의 최대 추출 및 항산화 활성을 위한 에탄올 농도를 선정하여 에탄올 추출물을 얻은 이후 용매의 극성차이에 의한 분획을 하였다. 커피체리 펄프의 항산화 성분으로는 총 페놀성 화합물 함량과 총 플라보노이드 함량을 비교 분석했을 때, 80% 에탄올 농도에서 각각 46.55 mg GAE/g과 113.42 mg QE/g으로 가장 높게 나타났다. 커피체리 펄프 80% 에탄올 추출물(CEE)을 이용해 n-hexane, chloroform, ethyl acetate, n-butanol 순으로 용매 분획을 진행하여 회수된 분획에 대한 항산화 성분 함량을 비교하였다. 각 분획물 중 총 페놀성 화합물 함량과 총 플라보노이드 함량은 ethyl acetate 분획물(CEF)이 가장 높아 각각 627.50 mg GAE/g과 1,632.31 mg QE/g이었으며, DPPH 라디칼 소거능 및 ABST 라디칼 소거능에 대한 IC50값이 각각 0.17 mg/mL와 0.34 mg/mL로 분획물 중 가장 낮아 항산화 활성이 높게 평가되었다. CEE에 함유된 페놀성 화합물을 HPLC system으로 분석한 결과 3-chlorogenic acid 15.05 mg/g, 4-chlorogenic acid 0.61 mg/g, 5-chlorogenic acid 1.06 mg/g, 3,4-dicaffeoylquinic acid 0.26 mg/g, 4,5-dicaffeoylquinic acid 1.22 mg/g, 3,5-dicaffeoylquinic acid 20.10 mg/g, caffeic acid 0.02 mg/g이었으며, CEF의 경우 3-chlorogenic acid 62.75 mg/g, 4-chlorogenic acid 1.47 mg/g, 5-chlorogenic acid 4.01 mg/g, 3,4-dicaffeoylquinic acid 7.26 mg/g, 4,5-dicaffeoylquinic acid 34.82 mg/g, 3,5- dicaffeoylquinic acid 72.26 mg/g, caffeic acid 0.66 mg/g으로 산출되었다. 특히, CEF에 함유된 3-chlorogenic acid와 3,5-dicaffeoylquinic acid의 함량이 CEE보다 3배 이상 증가하여 항산화 성분의 함량과 항산화 활성에 영향을 주었을 것으로 예측되었다. CCP가 함유된 MRS broth에 cinnamoyl esterase 유전자를 보유한 것으로 알려진 유산균을 접종하여 발효시킨 발효물에 대한 페놀성 화합물의 조성변화를 분석한 결과 CQA 및 DCGA의 함량은 감소했으나, Lactobacillus paracasei JBLAB1402 및 L. rhamnosus GG를 이용하여 발효했을 때, caffeic acid의 함량이 증가하는 것을 확인하였다. 이 결과를 토대로 커피체리 펄프에 함유된 페놀성 화합물의 조성과 유산균 발효를 통한 생체이용률 증가 기능성 소재의 확보에 관한 연구의 기초자료로 활용될 것으로 기대된다.

Chlorogenic acid modulates the ubiquitin–proteasome system in stroke animal model

Murad-Ali Shah,강주빈,고필옥 한국실험동물학회 2022 Laboratory Animal Research Vol.38 No.4

Background: Chlorogenic acid, a phenolic compound, has potent antioxidant and neuroprotective properties. The ubiquitin–proteasome system is an important regulators of neurodevelopment and modulators of neuronal function. This system is associated with neurodevelopment and neurotransmission through degradation and removal of damaged proteins. Activation of the ubiquitin–proteasome system is a critical factor in preventing cell death. We have previously reported a decrease in the activity of the ubiquitin–proteasome system during cerebral ischemia. This study investigated whether chlorogenic acid regulates the ubiquitin–proteasome system in an animal stroke model. In adult rats, middle cerebral artery occlusion (MCAO) surgery was performed to induce focal cerebral ischemia. Chlorogenic acid (30 mg/kg) or normal saline was injected into the abdominal cavity 2 h after MCAO surgery, and cerebral cortex tissues were collected 24 h after MCAO damage. Results: Chlorogenic acid attenuated neurobehavioral disorders and histopathological changes caused by MCAO damage. We identified the decreases in ubiquitin C-terminal hydrolase L1, ubiquitin thioesterase OTUB1, proteasome subunit α type 1, proteasome subunit α type 3, and proteasome subunit β type 4 expression using a proteomics approach in MCAO animals. The decrease in these proteins was alleviated by chlorogenic acid. In addition, the results of reverse transcription-polymerase chain reaction confirmed these changes. The identified proteins were markedly reduced in MCAO damage, while chlorogenic acid prevented these reductions induced by MCAO. The decrease of ubiquitin–proteasome system proteins in ischemic damage was associated with neuronal apoptosis. Conclusions: Our results showed that chlorogenic acid regulates ubiquitin–proteasome system proteins and protects cortical neurons from neuronal damage. These results provide evidence that chlorogenic acid has neuroprotective effects and maintains the ubiquitin–proteasome system in ischemic brain injury.

Identification of proteins regulated by chlorogenic acid in an ischemic animal model: a proteomic approach

Murad Ali Shah,강주빈,고필옥 한국실험동물학회 2023 Laboratory Animal Research Vol.39 No.2

Background: Cerebral ischemia is a serious neurological disorder that can lead to high morbidity and mortality. Chlorogenic acid is a polyphenol compound with antioxidant that can regulate proteins in cerebral ischemia. Middle cerebral artery occlusion (MCAO) surgery was performed to induce ischemic brain injury and was maintained for 24 h. Chlorogenic acid (30 mg/kg) or vehicle was administrated into the peritoneal cavity 2 h after MCAO surgery. The cerebral cortical tissues were collected for further study and a proteomic approach was performed to identify the proteins changed by chlorogenic acid in the MCAO animals. Results: We found that chlorogenic acid alleviated in changes in adenosylhomocysteinase, glycerol-3-phosphate dehydrogenase, eukaryotic translation initiation factor 4A-II, apolipoprotein A-I, and mu-crystallin. These proteins were reduced in MCAO animals with vehicle, and these reductions were attenuated by chlorogenic acid treatment. The mitigation of this reduction by chlorogenic acid was confirmed by the reverse transcription PCR technique. These proteins are associated with energy metabolism, protein synthesis, inflammation, and physiological metabolism. They are involved in the neuroprotective effect of chlorogenic acid. These results showed that chlorogenic acid alleviates the neurological disorders caused by MCAO and regulates the expression of proteins involved in neuroprotection. Conclusions: Therefore, our findings provide evidence that chlorogenic acid plays a neuroprotective role in stroke animal models by controlling specific proteins.

Identification of regulated proteins of the cerebral cortex differentially expressed by chlorogenic acid in middle cerebral artery occlusion animal model

Murad-Ali Shah,Ju-Bin Kang,Dong-Ju Park,Phil-Ok Koh 한국실험동물학회 2021 한국실험동물학회 학술발표대회 논문집 Vol.2021 No.7

Stroke is the global cause of death and permanent disability. Chlorogenic acid has antioxidant, and anti-apoptotic properties. In the current study, we investigated antioxidant and anti-apoptotic properties of chlorogenic acid against middle cerebral artery occlusion (MCAO)-induced cerebral ischemia in adult rats. Rats were injected intraperitoneally with chlorogenic acid (30 mg/kg) or vehicle 2 h after ischemia. After 24 h, cerebral cortex tissues were separated and reactive oxygen species (ROS) and malondialdehyde (MDA) assay, and histopathological analysis were performed. Two-dimensional gel electrophoresis and mass spectrometry techniques were performed to identify protein changes by chlorogenic acid. Results showed a significant increase in both ROS and MDA levels, histopathological lesions, and a high increase in terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells in the ischemic cerebral cortex. Conversely, chlorogenic acid-treatment significantly reduced these changes in MCAO-induced rats. Moreover, two-dimensional gel electrophoresis and western blot analysis showed significant changes by MCAO surgery. Expression levels of parvalbumin, isocitrate dehydrogenase 1, peroxiredoxin-2, and protein phosphatase 2A subunit A were decreased in the MCAO-induced cortex. However, these decreases were alleviated by chlorogenic acid-treatment. These proteins plays important role in calcium regulation, signal transduction pathways, metabolism, and oxidative stress. These results suggest that chlorogenic acid has neuroprotective properties by regulating of parvalbumin, isocitrate dehydrogenase 1, peroxiredoxin-2, and protein phosphatase 2A subunit A against cerebral ischemic damage. These results further suggest that chlorogenic acid can be used as a neuroprotective agent against diseases related to stroke by regulating various protein expressions. This research was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (NRF-2018R1D1A1B07044074).

Responses of the phenolic compounds of Zea mays under heavy metal stress

Kisa, Dursun,Elmastas, Mahfuz,Ozturk, Lokman,Kayir, Omer The Korean Society for Applied Biological Chemistr 2016 Applied Biological Chemistry (Appl Biol Chem) Vol.59 No.6

Heavy metal toxicity is one of the major abiotic stresses caused by physiological and biochemical changes. Plants have evolved various phytochemical defense mechanisms to cope with this abiotic stress conditions. Phenolic compounds are one of the stress responses and have multiple roles in respect to adaptation of plants to the environment. In the present study, we aimed to evaluate the differential accumulation of various phenolics with HPLC in the leaves of corn exposed to increasing heavy metal doses in the plant growth medium. The application of Cd, Cu, and Pb increased the total phenolics in all treatments compared to control groups. Chlorogenic acid and rutin were the main phenolic compounds in respect to quantifying. However, the contents of caffeic acid, ferulic acid, and vanillic acid were comparatively lower than chlorogenic acid and rutin in all samples. The content of chlorogenic acid significantly increased and rutin slightly increased in the treatment of the heavy metals. The levels of caffeic acid and ferulic acid significantly decreased in all exposures of heavy metals compared to control groups. The content of vanillic acid changed according to heavy metal types and doses in the leaves of corn, and the low doses of Pb and Cd increased the level of vanillic acid. We show that there is a positive correlation with the total phenolic content and chlorogenic acid when the corn is exposed to Pb. Moreover, there are negative correlations between total phenolic compound and caffeic acid, ferulic acid in the application of Cu and Cd.