UPLC를 이용한 lutein과 zeaxanthin의 분석법 검증 및 엽채류에서의 정량적 평가

김선아(편집자),김지선(Ji-Sun Kim) 한국식품과학회 2012 한국식품과학회지 Vol.44 No.6

본 연구에서는 UPLC를 이용하여 lutein과 zeaxanthin을 단시간에 동시 분석할 수 있는 새로운 방법을 개발한 후 분석법에 대한 평가를 수행하였으며, 확립된 방법으로 6종의 엽채류에서 lutein과 zeaxanthin을 정량 분석하였다. 분석 컬럼은 Acquity UPLCBEH C18(1.7 μm, 2.1×50 mm), 이동상 용매는 85% methanol을 사용하였으며, 검출파장은 450 nm, 이동상의 유속은 0.5 mL/min, 분석온도는 40℃, 시료주입량은 1.0 μL로 설정하여 분석하였다. 확립된 분석조건에서 lutein과 zeaxanthin의 피크머무름시간(RT)은 각각 4.35분과 4.56분이며, 표준용액의 피크머무름시간과 엽채류 시료들의 피크머무름시간은 일치하였다. lutein과 zeaxanthin에 대한 표준검정곡선은 1-150 μg/mL 농도범위에서 상관계수(r²)는 0.9968 이상의 양호한 직선성을 나타내어 분석에 적합함을 알 수 있었으며, 검출한계는 lutein과 zeaxanthin이 각각 1.7 μg/mL과 2.1μg/mL, 정량한계는 lutein과 zeaxanthin이 각각 5.1 μg/mL과 6.3 μg/mL로 설정되었다. Lutein과 zeaxanthin의 회수율은 각각 95.76-105.13%와 91.75-103.24% 범위를 보였다. Lutein의 일내 RSD값은 4.65-7.33%, 일간 RSD값은 4.04-10.69%이었으며, zeaxanthin의 일내 RSD값은 5.41-7.31%, 일간 RSD값은 3.79-9.82%이었다. 확립된 분석법으로 한국인이 많이 섭취하는 녹색과 엽채류인 부추, 취나물, 원추리, 참나물, 돌나물 시금치에서 lutein과 zeaxanthin을 분석한 결과 lutein 함량은 시금치와 취나물이 3.97±0.10 mg/100 g fw, 4.06±0.24 mg/100 g fw로 가장 높았으며, 참나물과 원추리가 3.39±0.43 mg/100 g fw, 3.39±0.15 mg/100 g fw로 높았다. Zeaxanthin의 경우 취나물과 원추리가 각각 0.95±0.00 mg/100 g fw와 0.96±0.01 mg/100 g fw로 가장 높았다. 본 연구에서 확립된 UPLC 분석법은 lutein과 zeaxanthin을 신속하고 효과적으로 동시 분석하는데 이용될 수 있을 것이며, 한국인이 많이 섭취하는 엽채류 중 시금치와 취나물은 lutein과 zeaxanthin의 좋은 식이 급원일 뿐 아니라, 꾸준히 섭취하면 눈건강에 도움을 줄 수 있을 것으로 사료된다. The objective of this research is to present method development and validation for the simultaneous determination of lutein and zeaxanthin using ultra performance liquid chromatography (UPLC). Also, rapid quantification was performed on six green leafy vegetables (Allium tuberosum, Aster scaber, Hemerocallis fulva, Pimpinella brachycarpa, Sedum sarmentosum and Spinacia oleracea) that are commonly consumed in Korea. Separation and quantification were successfully achieved with a Waters Acquity BEH C18 (50×2.1 mm, 1.7 μm) column by 85% methanol within 5 min. Two compounds showed good linearity (r²>0.9968) in 1-150 μg/mL. Limit of detection (LOD) and quantification (LOQ) for lutein and zeaxanthin were 1.7 and 5.1 g/mL and 2.1 and 6.3 g/mL, respectively. The RSD for intra- and inter-day precision of each compound was less than 10.69%. The recovery of each compound was in the range of 91.75-105.13%. Aster scaber and Spinacia oleracea contained significantly higher amounts of lutein (4.06±0.24 and 3.97±0.10 mg/100 g of fresh weight), respectively.

Bioavailability of lutein in corn distillers dried grains with solubles relative to lutein in corn gluten meal based on lutein retention in egg yolk

Shin, Hye Seong,Kim, Jong Woong,Lee, Dong Gu,Lee, Sanghyun,Kil, Dong Yong John Wiley Sons, Ltd 2016 Journal of the Science of Food and Agriculture Vol.96 No.10

<P>BACKGROUND: Dietary lutein and its food sources have gained great attention due to its health-promoting effects on humans, especially for certain eye diseases. However, relative bioavailability (RBV) of lutein among lutein-rich feed ingredients that lead to lutein-enriched egg production has not been determined. Thus, the RBV of lutein in corn distillers dried grains with solubles (DDGS) as compared to lutein in corn gluten meal (CGM) was evaluated based on lutein retention in egg yolk. RESULTS: Increasing inclusion levels of DDGS or CGM in diets increased (linear, P<0.01) Roche colour score and lutein concentrations of egg yolk without affecting laying performance. Multiple regression analysis revealed that the bioavailability of lutein in DDGS was less (P<0.05) than that of lutein in CGM, with the RBV of lutein in DDGS being 61.6% when the bioavailability of lutein in CGM was assumed to be 100% for lutein retention in egg yolk. CONCLUSION: The results of the present experiment indicate that the DDGS can be a potential ingredient for laying hens to improve egg yolk colour and lutein concentrations of egg yolk although lutein in DDGS is less bioavailable than lutein in CGM. (C) 2015 Society of Chemical Industry</P>

Preparation, characterization, and properties of lutein block polyethylene glycol copolymer loading with lutein nanoparticles

Peng Liu,Xiaoyu Bai,Xingtong Gao,Kai Liu,Aixiang Li,Zijian Lyu,Qiuhong Li 한국고분자학회 2023 Macromolecular Research Vol.31 No.3

In this paper, lutein block polyethylene glycol (lutein-b-PEG) copolymers were synthesized by hydrophilic modification of lutein with carboxylated polyethylene glycol (CT-PEG). The unreacted lutein was loaded into micelles formed by self-assembly of block copolymers to prepare composite nanoparticles. The results of FT-IR, 1H NMR, gel permeation chromatography (GPC), UV–vis and critical micellar concentration (CMC) showed that the double terminal carboxyl group ratio of CT-PEG reached 50.31%, and lutein-b-PEG was successfully synthesized without destroying the structure of lutein. Compared with lutein, the retention rate of lutein composite nanoparticles increased from 4.32 to 81.3% after 30 days of storage in the dark. In addition, the saturation solubility and bioaccessibility of lutein nanoparticles were increased 35 times and 5.2 times, respectively, due to micellar formation and improved water solubility. These findings indicated that the lutein composite nanoparticles modified with PEG significantly improved the chemical stability, water solubility, and bioaccessibility of lutein.

Effect of dietary lutein on the egg production, fertility, and oxidative injury indexes of aged hens

N. Liu,X. Ji,Z. Song,X. Deng,J. Wang Asian Australasian Association of Animal Productio 2023 Animal Bioscience Vol.36 No.8

Objective: The present study aimed to investigate the effect of dietary lutein on egg production, follicles, reproductive hormones, fertility, hatchability, and oxidative injury indexes of hens. Methods: Treatments consisted of a control diet (CON) and three lutein-supplementing diets at 25 (L1), 50 (L2), or 75 (L3) mg/kg of diet. Egg production was measured using 576 Arbor Acres breeder hens at 61 to 65 wk and follicles grades, reproductive hormones, fertility, hatchability, tissue lutein contents, and oxidative injury indexes were determined at 65 wk. Results: The results showed that at 65 wk, lutein- supplementing diets increased (p<0.05) egg production, follicular grades, fertility, hatchability, estradiol (E2), luteinizing hormone, progesterone (PROG), lutein content in the serum and yolk, compared to CON. L2 and L3 showed more pronounced (p<0.05) effects on egg production, PROG, and yolk lutein content than L1. With the increase of lutein doses from 25 to 75 mg/kg, there were linear increases (p<0.05) in egg production, lutein content, and PROG, and a quadratic trend (p<0.05) in E2. For the oxidative injury products, lutein-supplementing diets decreased (p<0.05) malondialdehyde (MDA) and protein carbonyl (PCO) in the serum, MDA and 8-hydroxy 2 deoxyguanosine (8-OHdG) in the yolk. There were linear decreases (p<0.05) in 8-OHdG in the serum, MDA, PCO, and 8-OHdG in the yolk, a quadratic trend (p<0.05) on serum 8-OHdG. Conclusion: It is concluded that lutein supplementation can improve egg production and fertility by beneficially regulating reproductive hormones and oxidative status in aged hens.

Comparison of static and dynamic in vitro digestion models to estimate the bioaccessibility of lutein in lutein-rich foods

이은하,차광현,Trang Thi Vuong,김상민,판철호 한국응용생명화학회 2018 Applied Biological Chemistry (Appl Biol Chem) Vol.61 No.4

This study aimed to determine the bioaccessibility of lutein in lutein-rich food, using static and dynamic models of in vitro gastrointestinal digestion. Here, kale powder (KP) and lutein supplement (LS) were used as representative lutein-rich foods. The bioaccessibility of lutein from KP did not considerably differ between static (59.92%) and dynamic (56.08%) digestion. Bioaccessibility was consistently maintained at the same level during dynamic digestion. The amount of lutein released from the LS during dynamic digestion was five times higher than that released during static digestion (67.88 vs 12.34%). The results showed that (a) bioaccessibility of lutein was affected by various factors such as food source, solid:liquid ratio, and interaction with dietary components, and (b) dynamic digestion should be suitable for evaluating the bioaccessibility of lutein in high-fat foods.

Lutein과 Apocarotenoic Acid Ethyl Ester 급여가 계육의 저장중 지질 산화에 미치는 영향

민병진,김혜정,강창기,이성기 한국축산식품학회 2003 한국축산식품학회지 Vol.23 No.3

육계사료에 xanthophylls의 첨가가 닭고기의 지질산화에 미치는 영향을 구명하기 위하여 실시하였다. 사료에 lutein과 apocarotenoic acid ethyl ester(Apo-ester)를 각각 10 ppm과 20 ppm 첨가하여 브로일러를 6주간 사육하였다. 도계후 가슴육과 다리육으로 분리하여 3$^{\circ}C$에서 9일, -18$^{\circ}C$에서의 4개월간 실험을 수행하였다. 냉장저장중 모든 계육의 pH는 증가하였고 xanthophylls 급여육이 대조구보다 높았다. 저장중 TBARS와 POV는 가슴육보다 다리육에서 더 높았다(p<0.05). Xanthophylls 급여육은 대조구에 비해 지질산화가 억제되었고(p<0.05) 가슴육보다는 다리육에서, 냉장기간보다는 냉동기간중에 억제효과가 더 높았다. 첨가농도별 효과를 보면 10ppm보다는 20 ppm 급여육에서 지질산화가 더 억제되었다. Lutein과 apo-ester 급여구간 유의차는 없었으나(p>0.05), lutein 20 ppm 급여육이 가장 높은 항산화 효과를 나타내었다. 따라서, 육계사료에 xanthophylls의 두 종류인 lutein과 apo-ester를 첨가하면 닭고기의 지질산화를 지연시킬 수 있다는 것을 알 수 있었다. This study was carried out to investigate the influence of dietary xanthophylls(lutein and apocarotenoic acid ethyl ester) supplementation on the antioxidation of broiler meat. The broilers fed with 10 ppm or 20 ppm xanthophylls were raised for 6 weeks and then slaughtered. The broiler meats were stored at 3$^{\circ}C$ for 9 days and frozen at -18$^{\circ}C$ for 4 months until analysis, respectively. The pH of all treatments significantly(p<0.05) increased during the storage periods. The pH of the thigh was higher than that of the breast. TBARS (thiobarbituric acid reactive substance) and POV(peroxide value) were higher in thigh than breast. All meats from broiler fed with lutein and apocarotenoic acid ethyl ester(apo-ester) had greater antioxidant properties during the storage period than control meat(p<0.05). Antioxidant activity of dietary xanthophylls supplementation was more effective in thigh than breast, and in broiler meats during frozen storage than chilled storage. The higher concentration of xanthophylls in feed, the more inhibition of lipid oxidation in meat during storage. The meat from broiler fed with 20 ppm of lutein showed the highest antioxidant property during both refrigerated and frozen storage although there was no significant difference between lutein and apo-ester(p>0.05). Consequently, this results indicated that the antioxidant activity of dietary xanthophylls(lutein and apocarotenoic acid ethyl ester) supplementation was more effective.

Lutein Modulates Th2 Immune Response in Ovalbumin-Induced Airway Inflammation

Jun Young Song(송준영),Chang-Min Lee(이창민),Min Ki Lee(이민기) 한국생명과학회 2012 생명과학회지 Vol.22 No.3

Lutein은 식물에서 발견되는 carotenoid 계열에 속하는 물질로 항산화 기능을 가지고 있는 것으로 널리 알려져 있지만, 호흡기 질환과 관련하여 아직까지 lutein의 효능과 작용 기작이 잘 알려져 있지 않다. Ovalbumin (OVA)으로 유도한 천식(asthma) 생쥐모델에서 lutein은 기도 과민성을 억제하였고, 기관지 폐포 세척액에서 OVA의 감작에 의하여 증가한 각종 염증성 지표들을 감소시켰다. 또한, OVA의 감작에 의하여 증가한 제2형 협조 T 세포(Th2 cell)의 증가된 반응을 약화시키는 결과를 볼 수 있었다. 본 실험에서 lutein이 ovalbumin (OVA)으로 유도한 천식 생쥐 모델에서 제 2형 협조 T세포의 싸이토카인과 유전자 발현을 조절할 수 있는 면역약리학적 기능을 할 수 있는 물질로서의 가능성이 있음을 확인할 수 있었다. The general term flavonoids is often used to categorize a family of natural compounds that are highly abundant in all higher plants, and which in recent years have attracted scientific interest as therapeutics. Lutein is a xanthophyll and one of 600 known naturally occurring carotenoids. It is found in green vegetables such as spinach and kale, and has been demonstrated to exert anti-inflammatory activities. However, its anti-allergic effect in the Th1/Th2 immune response is poorly understood. In this study, we attempt to determine whether lutein regulates inflammatory mediators in an ovalbumin (OVA)-induced murine asthma model. To address this, mice were sensitized and challenged with OVA, and then treated with lutein before the last OVA challenge. Administration of lutein significantly suppressed the OVA-induced airway hyper-responsiveness. It also resulted in a significant alleviation of the infiltration of inflammatory cells into the bronchoalveolar lavage. Additionally, lutein attenuated the increased expression of Th2 responses in OVA-challenged mice. These results demonstrate that lutein is a potent inhibitor that reduces Th2 immune responses. Furthermore, they show that the immunopharmacological function is mediated by a pathway that involves and is regulated by Th2 immune response.

Expression of Sloan-Kettering Virus in the Corpus Luteum in the Rat Ovary

Eun-Ji Park,Dong Hun Kim,Soo Bong Park,Yeoung-Gyu Ko,Sung Woo Kim,Yoon Jung Do,Hyun Kim 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s

Sloan-Kettering virus gene product of a cellular pro-oncogene c-Ski is an unique nuclear pro-onco protein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea(CL) formation to predict the possible involvement of Ski in luteinization. In addition, to examine whether the initiation of luteinization with luteinizing hormone(LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone(LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum(CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

Expression of Ski in the Corpus Luteum in the Rat Ovary

김현,Takashi Matsuwaki,Keitaro Yamanouchi,Masugi Nishihara,양보석,고응규,김성우 사단법인 한국동물생명공학회 2011 한국동물생명공학회지 Vol.26 No.4

Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea (CL) formation to predict the possible involvement of Ski in luteinization. In addition, we performed to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

루테인 공급원이 난황내 루테인 축적률에 미치는 영향

강근호,김상호,강환구,김동욱,김지혁,김현수,황보종,유동조,나재천,방한태,강보석,최희철,서옥석 한국가금학회 2008 한국가금학회 정기총회 및 학술발표회 Vol.25 No.-

This experiment was conducted to investigate the effect of dietary supplementation of the various materials contained lutein on the lutein accumulation in egg yolk. The laying hens were fed corn-soybean meal based diets containing 0, 0.6, 1, 3, 5 or 10 ppm of lutein. The lutein content of egg yolk was linear increased by increasing the supplementation level of lutein. The spinach 10 treatment, lutein content was the highest than that those treatments from egg yolk. But the efficiency of lutein accumulation in egg yolk was significantly higher in lutein extract treatment as compared to that of raw materials treated groups. The results of this experiment suggested that the dietary supplementation of lutein extract was more effective than that of raw materials for producing the lutein enriched eggs.