산초나무(Zanthoxylum schinifolium) 종자는 동아시아에서 향신료 및 약용 자원으로 널리 활용되고 있으나, 화학적으로 규정된 지표물질을 기반으로 한 체계적인 품질평가 기준은 아직 확립되지 않았다. 본 연구에서는 HPLC–DAD와 LC–MS/MS 분석을 통합적으로 적용하여 산초나무 종자의 품질평가를 위한 지표물질로서 hesperidin을 확인하고 검증하였다. 산초 종자의 메탄올 조추출물에 대해 크로마토그래피 분석을 수행한 결과, 주요 플라보노이드 성분들은 UV 흡수 특성과 음이온 ESI–MS/MS 단편화 패턴을 통해 동정되었다. 특히 m/z 609.6[M–H]–의 주요 전구체 이온은 hexose (162Da)와 rhamnose (146Da)의 순차적 중성손실을 거쳐 각각 m/z 463.4 및 301.3의 단편 이온을 생성하였으며, 이러한 단편화 거동은 표준물질 hesperetin-7-O-rutinoside (hesperidin)의 스펙트럼과 완전히 일치하였다. 이후 정량분석을 위해 확립된 HPLC–DAD 분석법은 우수한 직선성(R2 > 0.999), 높은 민감도(LOD 및 LOQ가 허용 범위 내), 그리고 재현성과 정밀도를 나타내어 분석법의 신뢰성을 확인하였다. 또한 여러 배치의 산초나무 종자 시료에서 hesperidin 함량이 일관되게 검출되어, 본 성분이 품질관리 지표물질로서 적합함을 확인하였다. 종합적으로, HPLC–MS/MS 기반 구조 규명과 정량 분석 결과는 hesperidin이 산초나무 종자의 주요 플라보노이드 배당체이며, 화학적 표준화 및 품질평가를 위한 적절한 지표성분임을 명확하게 보여준다. 본 연구는 산초나무 유래 기능성 소재의 산업적 활용과 표준화, 품질 보증을 위한 분석 기반을 제공하는 데 중요한 의의가 있다.

To produce an intestinal immunomodulatory beverage containing Centella asiatica extract (CAE), three types of CAE-added beverage prototypes were prepared, and their immunomodulatory activities and marker compounds were analyzed. As a result of the cytotoxicity assessment, all the beverages did not show significant toxicity compared to the control group. Next, the immunomodulatory activities of the beverage prototype were evaluated using the inflammatory model of IL-1β-induced intestinal epithelial cell line. All the samples significantly reduced the production of IL-6, IL-8, and MCP-1 in a CAE concentration-dependent manner. In addition, CAE-added beverages inhibited NO, IL-6, and IL-12 production in LPS-induced RAW 264.7 cells. When the major triterpenoids, as marker compounds for the production of CAE-added beverages, were analyzed by HPLC-DAD, only asiaticoside was detected beyond the limit of quantification, while madecassoside, madecassic acid, and asiatic acid were not detected. The amounts of asiaticoside in CAE-added beverage prototypes were confirmed in No. 1 (19.39 μg/mL), 2 (19.25 μg/mL), and 3 (19.98 μg/mL). In conclusion, the results of this study suggested that CAE-added beverage prototypes induced immunomodulatory effects in the intestinal inflammatory cell line models and asiaticoside could be used as a marker compound for CAE-added beverage production.

Centella asiatica (C. asiatica) has been widely used in food, cosmetics, and pharmaceutical industry as a functional material. In a previous study, we have investigated not only pharmacological effects such as antioxidative and anti-inflammatory effects, but also analyzed various functional ingredients. In this study, triterpenoids were analyzed using HPLC-DAD to determine marker compounds among functional ingredients. When triterpenoids were analyzed, asiaticoside from C. asiatica was determined as an optimal marker compound. Next, specificity, linearity, limited of detection (LOD), limited of quantification (LOQ), precision, accuracy, and range were evaluated using HPLC-DAD to determine asiaticoside contents in C. asiatica juice and extracts. The specificity was elucidated by chromatogram and retention time using an established analytical method. The coefficient of correlation obtained was 0.9996. LOD was 4.99 μg/mL and LOQ was 15.12 μg/mL. Intra- and inter-day precision of asiaticoside were determined to be 0.48~1.68% and 0.08~1.09%, respectively. Furthermore, the recovery rate of asiaticoside was 98.88% and the analytical range of Field-70E was determined to be 0.625~10 mg/mL. As a results of evaluating ABTS, DPPH, and FRAP antioxidative effect, Field-70E showed potent antioxidant activities. Results of this study could be used as basic data for quality standardization of C. astiatica juice and extracts.

복분자를 개별인정형 건강기능식품 기능성 원료로 개발하기 위하여 지표성분 표준화를 위한 ellagic acid의 분석법 설정과 분석법에 대한 검증(validation)을 실시하였다.그 결과 ellagic acid의 검량선은 R2=0.9999으로 좋은 선형성을 보였으며, 검출한계 0.6µg/mL, 정량한계는 1.9µg/mL였다. 회수율을 측정한 결과에서는 89.0-100.0%로 나왔고, 상대표준편차(RSD)는 0.05-0.14%를 보였으며, 일내와일간 분석에서 RSD는 각각 0.28-1.96%와 0.69-2.49%로나왔다. 따라서 본 연구를 통해 복분자 추출물의 지표물질인 ellagic acid의 분석법은 적합한 시험법임이 검증되었다.

Background : This study was carried out to investigate the possibility of cosmetics materials by comparing growth characteristics, photosynthetic rate and major functional components of Rosa multiflora and Perilla frutescens at different altitudes. Methods and Results : This experiment is being carried out in April 2018 in Namwon (500 m above sea level) and Iksan (15 m above sea level) in Jeollabuk-do. The growth characteristics of R. multiflora were investigated at the end of May. Flowers were collected at this time and used as samples for functional analysis. The growth characteristics of P. frutescens were investigated in the middle of August and the ground part was collected at this time and used as a sample for functional analysis. Photosynthetic rates were measured using LCpro+ (ADC, UK). The marker compound were investigated and analyzed using HPLC Alliance e2695 and 2998 PDA detector (Waters, USA). Photosynthetic rate (based on 1,600 μ mole of light intensity) was measured in mid-June as follows. The R. multiflora showed 9.8 μ mole․CO2/㎡/s in Iksan and 7.9 μmole․CO2/㎡/s in Namwon. The P. frutescens showed 15.0 μmole․CO2/㎡/s in Iksan and 8.8 μmole․CO2/㎡/s in Namwon. Overall, Photosynthetic rate was higher in Iksan. As a result of analyzing 18 kinds of marker compound, gallic acid and astragalin were found in R. multiflora, caffeic acid and rosmarinic acid were found in P. frutescens. Gallic acid and Astragalin of R. multiflora showed 5.4 ㎎/g and 28.4 ㎎/g in Iksan and 3.2 ㎎/g and 21.6 ㎎/g in Namwon, respectively. Caffeic acid and rosmarinic acid of P. frutescens were 2.7 ㎎/g and 49.7 ㎎/g in Iksan and 2.5 ㎎/g and 33.6 ㎎/g in Namwon, respectively. Conclusion : Comparing the yield of the harvesting parts by region, both R. multiflora and P. frutescens was higher in Namwon. As a result of quantitative analysis of four detected elements of gallic acid, astragalin, caffeic acid and rosmarinic acid, all four components were high in Iksan. It is considered that this is due to optical environment difference.

Background: In the present study, we established an HPLC (high performance liquid chromatography)-analysis method for the determination of marker compounds as a part of the material standardization for the development of health-functional foods from Salvia plebeia R. Br. extract. Methods and Results: The quantitative determination method of hispidulin as a marker compound was optimized by HPLC analysis using a YMC hydrosphere C18 column with a gradient elution system. This method was validated using specificity, linearity, accuracy, and precision tests. It showed a high linearity in the calibration curve with a coefficient of correlation (r2) of 0.999995. The method was fully validated, and was sensitive, with the limit of detection (LOD) at 0.09 ㎍• ㎖−1 and limit of quantification (LOQ) at 0.27 ㎍• ㎖−1. The relative standard deviation (RSD) values of the data from intra- and inter-day precision were 0.05 - 0.22% and 0.32 - 0.42%, respectively, and the intra- and inter-day accuracy of hispidulin were 99.5 - 102.3% and 98.8 - 101.5%, respectively. The average content of hispidulin in Salvia plebeia R. Br. extract was 3.945 ㎎• g−1 (0.39%). Conclusions: These results suggest that the developed HPLC method is very efficient, and that it could contribute to the quality control of Salvia plebeia R. Br. extracts as a functional ingredient in health functional foods.