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A kinetic assay was carried out in order to compare the ability of detection for prekallikrein activator(PKA) in plasma-derived products with that of an endpoint assay and a commercial method. Using these methods, 9 human albumin preparations were assayed and compared to each other. The coefficient of variation between the Kinetic assay and the end point assay was found within 6.6% and this result showed that two methods were highly correlative and the end point assay could act as a replacement of the kinetic assay. Another important goal of this study was to investigate the reproducibility among laboratories on the kinetic assay. A collaborative study was performed to validate the kinetic method with intra and inter assays. The coefficient of variation for the intra assay of each laboratory was less than 4% and that for between individuals in the inter assay was 4.1%. These results revealed that the kinetic assay showed good reproducibility. The contents of PKA in plasma-derived products were also determined by the kinetic assay. As a result, it was found that trace amounts of PKA were present in 32 human immunoglobulin preparations, however the average concentration of PKA in 171 albumin preparations was 5.8 IU/mL.

목적: 본 연구는 HPLC를 이용하여 작약 뿌리 추출물의 pentagalloylglucose을 지표물질로 설정하고 분석법을 확립하여 유효성 검 증을 하였다. 방법: Pentagalloylglucose 분석을 위해 C18컬럼을 이용하여 이동상 용매를 통해 HPLC 분석을 진행하였다. 분석법의 유효성 검증은 특이성, 직선성, 정확도, 정밀도, 검출한계 및 정량한계 등을 통해 분석법의 신뢰성을 검증하였다. 결과: 작약 뿌리 추출물과 pentaglloylglucose 표준용액 간의 HPLC 크로마토그램 및 UV spectrum의 비교를 통해 다르 물질들의 간섭 없이 피크가 분리된 것으로 특이성을 확인하였으며 상관계수(R2)는 0.9999로 매우 우수한 직선성을 관찰되었다. 검량선의 기울기 및 표준편차 를 이용한 검출한계는 0.92 μg/mL였고 정량한계는 2.72 μg/mL으로 확인되었다. 또한, 정확성 분석을 위해 서로 다른 농도 3개를 선정하여 확인한 결과 RSD가 약 0.12-0.50%로 확인되었다. 결론: 이상의 분석결과를 통해 확립된 작약 뿌리 추출물의 지표성분인 pentagalloylglucose의 HPLC 분석법은 적합한 시험법으로 검증되었으며 원료 표준화를 통해 기능성 원료 자료로 활용될 것으로 사료된다.

Purpose: This study established a high-performance liquid chromatography (HPLC) method to standardize pentagalloylglucose as a marker compound in Paeonia lactiflora (P. lactiflora) root extracts. Methods: The analytical procedure for standardizing pentagalloylglucose was optimized using HPLC on a C18 column with a gradient elution of 0.1% trifluoroacetic acid and acetonitrile. The method was validated for specificity, linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision. Results: The method demonstrated high linearity, as evident from a correlation coefficient R2 of 0.9999. The LOD and LOQ were 0.92 and 2.78 μg/mL, respectively. Recovery was determined to be in the range of 101.15 ± 0.49–102.30±0.13, with the relative standard deviation (RSD) values between 0.12%–0.50%, indicating high accuracy. Conclusion: The research lays a foundation for the production of functional foods or cosmetics using pentagalloylglucose derived from P. lactiflora root extract.

目的: 本研究采用HPLC方法建立芍药根提取物的五没食子酰葡萄糖作为指标，建立分析方法，并验证其有效 性。方法: 对于五没食子酰葡萄糖分析，使用C18色谱柱通过流动相溶剂进行HPLC分析。通过特异性，线性， 检测极限(LOD)，定量极限(LOQ)验证了分析方法的可靠性。结果: 通过对比芍药根提取物与五没食子酰葡萄糖 标准溶液的HPLC色谱图和紫外光谱，不受其他物质干扰，峰分离，确认了特异性，且线性良好，相关系数(R2) 为0.9999。使用校准曲线的斜率和标准差的检测限为0.92 μg/mL，并且定量限被确认为2.72 μg/mL。另外，选 择3个不同浓度进行精度分析，结果确认RSD约为0.12-0.50%。结论: 通过上述分析结果建立的芍药根提取物指 示成分五没食子酰葡萄糖的HPLC分析方法，经验证是一种合适的检测方法，有望通过原料标准化作为功能性原 料数据。

개별 인정형 건강기능식품 기능성 원료로 개발한 고흥 유자추출물의 표준화를 위해 지표성분으로 naringin을 설정하였으며, HPLC를 이용하여 지표성분 naringin의 분석법을 확립하고 그에 따른 유효성분 검정을 실시하고자 하였다. 유효성 검정 결과, 본 시험법에서 표준용액의 retention time과 고흥 유자 추출물의 retention time이 일치하는 것을 확인할 수 있었으며 동일한 spectrum을 나타내어 특이성을 확인하였다. 검량선의 상관계수($R^2$)는 0.9986으로 높은 직선성을 보였으며 검출한계는 0.0218 mg/L, 정량한계는 0.0661 mg/L로 설정되었다. Naringin의 회수율은 1 mg/mL에서는 95.75~98.25%, 0.5 mg/mL에서는 97.67~101.01%, 0.1 mg/mL에서는 97.33~104.64%, 0.05 mg/mL에서는 95.53~106.82%의 범위의 회수율을 얻었으며 intra-day에서의 정밀도(RSD)는 1.39~1.95%, inter-day에서는 0.17~1.49%의 정밀도를 나타내어 고흥 유자 추출물의 지표성분 naringin의 분석법은 적합한 시험법임을 확인하였다.

An HPLC method for determination of naringin was developed to standardize it as a marker compound in Goheung yuzu extract as a functional health food. Optimum results were obtained by C-18 column chromatography using solvent mixtures (A: 0.5% acetic acid, B: acetonitrile) as the stationary phase and mobile phase. The method was fully validated and sensitive with a limit of detection (LOD) of 0.0218 mg/L and limit of quantification (LOQ) of 0.0661 mg/L. The method showed high linearity (coefficient of correlation=0.9986) and high accuracy, as recovery rates of naringin at concentrations of 1, 0.5, 0.1, 0.05 mg/mL were in the ranges of 95.74~98.25%, 97.67~101.01%, 97.33~104.64%, and 95.53~106.82%, respectively. Intra-day and inter-day variation, which are measures of method precision, were 1.39~1.95% and 0.17~1.49%, respectively. Therefore, the method could be used without modification for determination of naringin as a marker compound in Goheung yuzu extracts.

This study aimed to establish a marker compound in roasted Astragalus membranaceus (AM) water extract and to validate its analytical methods. The roasting process significantly enhanced the isoflavone content in AM. Among the four isoflavones analyzed, calycosin 7-glucoside (C7G) emerged as the most abundant, with a concentration of 847.88 ㎍/g in the AM extract. Due to its concentration and representativeness, C7G was designated as the marker compound, and its analytical method was thoroughly validated. Specificity was confirmed by the consistent retention time of the C7G peak at 15.2 minutes across both the sample and the standard compound. High absorbance was recorded at UV wavelengths of 220, 250, and 260 nm. The method exhibited excellent linearity, with a correlation coefficient (R²) of 0.9999 across a concentration range of 0.2 to 50.0 ㎍/mL. The limits of detection and quantification were determined to be 0.029 ㎍/mL and 0.088 ㎍/mL, respectively. Precision assessments revealed intra-day and inter-day variations of 0.812% and 1.650%, respectively. Recovery tests yielded values ranging from 99.419% to 104.861%, with relative standard deviations between 1.152% and 2.215%. These results affirm that the analytical method for C7G is highly specific, linear, accurate, and precise. This validated method may serve as a valuable tool for the standardization of roasted AM water extract.

본 연구에서는 생물전환에 의한 품종별 대두 5종의 isoflavone 함량, 총페놀 함량, 항산화능 및 isoflavone의 분석법 검증을 시행하였다. HPLC를 이용한 분석법 검증 결과 isoflavone 6종의 표준용액과 우람(S-3) 비발효 및 발효 추출물의 retention time이 일치하는 것을 확인하였으며, spectrum 분석 결과에서도 같은 spectrum을 보여 특이성을 확인하였다. 직선성 확인 결과 isoflavone 6종의 검량선의 상관계수 값은 0.9998~1.0000으로 나타나 우수한 직선성을 보여주었다. 정밀성 측정 결과 일내 정밀도에서 0.22~0.88%, 일간 정밀도에서 0.34~1.00%의 높은 정밀도를 나타냈다. 회수율 측정 결과 daidzin은 100.55~110.21%, glycitin은 100.26~112.35%, genistin은 97.67~111.89%, daidzein은 100.95~110.43%, glycitein은 98.56~106.02%, genistein은 101.29~110.25%의 회수율을 나타내었다. Daidzin, glycitin, genistin의 검출한계는 각각 0.14, 0.16, $0.20{\mu}g/mL$로 측정되었으며, daidzein, glycitein, genistein의 검출한계는 각각 0.25, 0.29, $0.20{\mu}g/mL$로 나타났다. Daidzin, glycitin, genistin의 정량한계는 각각 0.44, 0.48, $0.60{\mu}g/mL$를 나타냈고 daidzein, glycitein, genistein은 각각 0.75, 0.89, $0.61{\mu}g/mL$로 나타났다. 생물전환에 의한 대두 발효 시 배당체인 daidzin, glycitin, genistein이 모두 비배당체인 daidzein, glycitein, genistein으로 전환됨을 확인할 수 있었다. 각각의 isoflavone을 합한 값인 total isoflavone 함량은 대두 우람(S-3) 발효물에서 $2,074.6{\mu}g/g$으로 가장 높은 함량을 나타냈으며 비발효 대두 새단백(S-5)에서 가장 낮은 $119.8{\mu}g/g$의 total isoflavone 함량을 나타내었다. 총페놀 함량은 진풍(S-4) 비발효물이 44.96 mg GAE/g으로 가장 높은 총페놀 함량을 보였다. 항산화능 측정 결과 FRAP는 대풍2(S-2) 발효물에서 0.22로 가장 높은 값을 나타냈으며 새단백(S-5) 비발효물에서 0.15로 가장 낮은 값을 나타내었다. ORAC 지수도 FRAP와 유사하게 새단백(S-5) 비발효물에서 $195.24{\pm}3.08{\mu}M\;TE/g$으로 가장 낮은 값을 보였으며 대풍 2(S-2) 발효물에서 $753.79{\pm}6.95{\mu}M\;TE/g$으로 가장 높은 값을 나타내었다.

The present study evaluated the validation method for isoflavone contents of fermented soybean extracts by bioconversion as well as their antioxidant activities. Our results show that the total isoflavone contents of non-fermented and fermented soybean extract ranged between 119.8 to $637.7{\mu}g/g$ and between 567.3 to $2,074.6{\mu}g/g$, respectively. Moreover, fermented soybean extracts had higher contents of isoflavone aglycones, including daidzein, glycitein, and genistein than non-fermented soybean extracts as well as lower contents of isoflavone glucosides such as daidzin, glycitin, and genistin. FRAP and ORAC values ranged between 0.15 to 0.22 and between 195.24 to $753.79{\mu}M$ Trolox equivalents/g in non-fermented and fermented soybean extracts, respectively. These results indicate that fermented soybean extracts had higher total isoflavone contents and antioxidant activities than non-fermented soybean extracts. Bioconversion process in this study may have the potential to produce isoflavone-enriched natural antioxidant agents with high added value from soybean matrices.

We propose a rigorous and practical methodology to evaluate the performance of symbolic manipulation program such as Mathematica, Maple, and Maxima. First, we demonstrate an inverse method to construct the benchmark problems of an initial value problems. The benchmark problems associated with the discrete version of the Chebyshev polynomials provide a rigorous and objective measure to evaluate the performance of symbolic manipulation programs. We compare three symbolic manipulation programs, which are Mathematica, Maple and Maxima, using this methodology. The computation time, the used memory and the perturbation terms are chosen for comparison parameters.

This study aimed to investigate the changes in the ${\gamma}$-aminobutyric acid (GABA) content of bitter melon (Momordica charantia L.) cultivated from different regions, with different harvest times and at various maturation stages. Methods for observing the changes in GABA content were validated by determining the specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), and precision and accuracy using the HPLC-FLD system. Results showed high linearity in the calibration curve with a coefficient of correlation ($R^2$) of 0.9999. The LOD and LOQ values for GABA were 0.29 and $0.87{\mu}g/mL$, respectively. The relative standard deviations for intra- and inter-day precision of GABA were less than 5%. The recovery rate of GABA was in the range of 98.77% to 100.50%. The average content of GABA was 0.93 mg/g and Cheongju showed highest GABA content of 1.88 mg/g. As the time of harvest increased from May to September, the GABA content decreased from 1.56 to 0.86 mg/g. Also, maturation of the bitter melon fruit was associated with a decreased in GABA content.

본 연구는 HPLC를 이용해 개별 인정형 건강기능식품 원료로 개발하기 위하여 곰취잎 추출물의 지표성분인 CA, 3,4-DCQA, 3,5-DCQA 및 4,5-DCQA의 동시분석법 설정과 분석법에 대한 검증을 실시하고자 하였다. 그 검증의 결과 표준액과 곰취잎 추출물의 HPLC 크로마토그램을 비교하여 피크가 분리됨을 확인하고 다른 물질의 간섭 없이 분리되었으며, 표준액의 피크 유지시간과 추출물의 피크 유지시간이 일치하였고 동일한 spectrum을 나타내었다. 또한 blank에서 표준액과 겹치는 피크가 없는 것으로 특이성을 확인하였다. 검량선의 상관계수는 CA, 3,4-DCQA, 3,5-DCQA 및 4,5-DCQA 각각 0.9999, 0.9999, 0.9999 및 0.9999로 모두 양호한 직선성을 보였으며, 직선상의 검출한계는 $3.0{\sim}13.2{\mu}g/mL$였으며 정량한계는 $9.2{\sim}39.8{\mu}g/mL$로 나타났다. 곰취 추출물 0.6, 1.2 및 1.8 mg/mL의 세 농도의 회수율은 98.96~101.81%였으며 RSD는 0.14~0.89%로 나타나 RSD 2.0% 이하로서 정확성이 있음을 알 수 있었다. 정밀성은 0.04~0.51%의 정밀도를, 실험실 내 정밀성에서는 0.02~0.90%의 정밀도를 나타내었고 intra-day에서의 정밀도는 0.07~0.63%, inter-day에서는 0.39~0.66%의 정밀도를 나타내어 곰취잎 추출물의 지표성분 CA, 3,4-DCQA, 3,5-DCQA 및 4,5-DCQA의 분석법은 적합한 시험법임이 검증되었다. 본 분석법이 곰취 추출물 개별 인정형 건강기능식품 원료 개발을 위한 기초자료로 활용될 것으로 사료된다.

An HPLC analysis method was developed for standard determinations of chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4,5-di-O-caffeoylquinic acid as functional health materials in Ligularia fischeri extract. HPLC was performed on a $C_{18}$ Kromasil column ($4.6{\times}250mm$, $5{\mu}m$ column) with a gradient elution of 0.1% (v/v) trifluoroacetic acid and acetonitrile at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analytes were detected at 330 nm. The HPLC method was validated in accordance with the International Conference on Harmonization guideline of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation for the four compounds were 3.0~14.6 and $9.2{\sim}44.4{\mu}g/mL$, respectively. Calibration curves showed good linearity ($r^2$ > 0.999), and the precision of analysis was satisfied (less than 0.9%). Recoveries of quantified compounds ranged from 98.96 to 101.81%. This result indicates that the established HPLC method is very useful for the determination of marker compounds in Ligularia fischeri leaf extracts.

본 연구진은 배박 추출물이 지방세포 대사를 저해함을 확인 하였으며, 배박을 건강기능식품 개별 인정형 소재로 개발하고자 하였다. 소재 개발에 앞서 배박 분석방법을 확립하고자 하였으며, 본 연구에서는 배박에서 확인된 성분인 caffeic acid 및 chlorogenic acid를 HPLC/UV system을 이용해 분석방법을 최적화하였다. 분석법이 타당한지를 검토하기 위하여 분석법 벨리데이션을 수행하였으며, 직선성, 일간 일내 정확성, 정밀성 검출한계 및 검량한계를 확인하였다. 본 시험분석법으로 배박 추출물 분석 시 지표물질이 다른 물질의 간섭 없이 안정되게 분석되는 것을 확인하였다. 따라서 본 분석방법은 배박 추출물 기능성 원료 표준화를 위한 시험법으로 적합한 것으로 판단되었다. 본 시험법에 따라 에탄올 투입량에 따른 배박 추출물 내의 caffeic acid 및 chlorogenic acid의 함량을 분석하였으며, 에탄올 투입에 따른 대상 성분의 추출율을 조절할 수 있음을 보여주었다. 따라서 본 연구를 통하여 확립된 분석법이 배박 추출물 개별 인정형 건강기능식품 기능성 원료 개발을 위한 기초 자료로 활용될 것으로 사료된다.

This study was conducted to establish an HPLC analysis method for determination of marker compounds as part of materials standardization for development of health functional food materials from pear pomace. The quantitative determination method of caffeic acid and chlorogenic acid as marker compounds of pear pomace extract (PPE) was optimized by HPLC analysis using a C18 column ($5{\times}250mm$, $5{\mu}m$) with a 0.2% elution gradient of acetic acid and methanol as the mobile phase at a flow rate of 0.8 mL/min and detection wavelength of 330 nm. The HPLC/UV method was applied successfully to the quantification of marker compounds in PPE after validation of the method with linearity, accuracy, and precision. The method showed high linearity of the calibration curve with a coefficient of correlation ($R^2$) of 0.9999, and limit of detection and limit of quantification were $1.14{\mu}g/mL$ (caffeic acid) and $1.61{\mu}g/mL$ (chlorogenic acid) as well as $4.9{\mu}g/mL$ (caffeic acid) and $4.9{\mu}g/mL$ (chlorogenic acid), respectively. Relative standard deviation values from intra- and inter-day precision were less than 3.1% (caffeic acid) and 4.0% (chlorogenic acid), respectively. Recovery rates of caffeic acid and chlorogenic acid at 12.5, 25, and $50{\mu}g/mL$ were 93.66~106.32% and 97.33~105.68%, respectively. An optimized method for extraction of caffeic acid and chlorogenic acid in PPE was established through diverse extraction conditions, and the validation indicated that the method is very useful for evaluation of marker compounds in PPE to develop a health functional food material.

The aim of this study was to select compounds for the standardization of fermented Kalopanax pictus Nakai (KP-F), to develop the analysis method using HPLC-PDA and to perform method validation. KP-F is a fermented powder developed to improve the original physiological activities and create a new functionality. Eleutheroside E, Acanthoside B, and Syringaresinol were selected as the standard compounds and developed our own method for simultaneous analysis. The analyte was isolated using C18 column with a gradient elution of 0.05 M phosphoric acid in water and methanol as the mobile phase at a flow rate of 1 mL/min and detected at 210 nm. As a result, all standard compounds showed good linearity with an $R^2$ (coefficient of correlation) of 1.000 and for the limit of detection range of $0.710{\sim}0.831{\mu}g/mL$, and the limit of quantification as $2.150{\sim}2.520{\mu}g/mL$. The precision was RSD (%) of less than 4.80%, while the accuracy was 4.70%>RSD (%) for the range 102.44~110.48%. In conclusion, the developed analysis method is suitable for the detection of Eleutheroside E, Acanthoside B, and Syringaresinol in KP-F.

Objectives: This study was performed to evaluate the analytical method for PAH metabolites in human urine using enzyme hydrolysis and solid-phase extraction coupled with LC-(ESI)-MS/MS technique. Methods: We employed HPLC tandem mass spectrometry techniques with appropriate pre-treatment for analysis of 16 OH-PAHs in human urine. Samples were hydrolysis by ${\beta}$-flucuronidase/Aryl sulfatase, and target compounds were extracted by solid-phase extraction with a strata-x cartridge. Cross-validation was performed between Eulji University and Green Cross laboratories with 200 human urine samples. Results: The accuracies were between 90.3% and 118.8%, and precisions (relative standard deviations) were lower than 10%. The linearity obtained was satisfying for the 16 OH-PAH compounds, with a coefficient of determination ($r^2$) higher than 0.99. The results of cross-validation at the two organizations were compared by ICC (interclass correlation coefficient) values. The cross-validation results were excellent or good for all compounds. Conclusion: An analytical method was validated for low nanogram levels of 16 OH-PAHs in human urine. Also, satisfying results were obtained for method validation such as accuracy, precision and ICC of cross-validation.

팬데믹 현상은 원격으로 문제를 해결할 수 있는 비대면 환경을 빠르게 발전시켰다. 하지만 급작스러운 비대면 환경으로 전환은 다양한 부분에서 새로운 보안 이슈들을 발생시켰다. 새로운 보안 이슈들 중 하나가 내부자에 의한 보안 위협이었고 이를 방어하기 위한 기술로 제로 트러스트 보안 모델이 다시 주목받게 되었다. SDP(Software Defined Perimeter) 기술은 다양한 보안 요소로 이루어져 있는데 이 중 디바이스 유효성 검증이라는 기술이 내부자의 사용 행위를 모니터링 하여 제로 트러스트 보안 모델을 실현할 수 있는 기술이다. 하지만 현재 SDP 명세서에는 디바이스 유효성 검증을 수행할 수 있는 기술이 제시되어 있지 않다. 따라서 본 논문에서는 SDP 환경에서 사용자 행위 모니터링을 통한 SVDD 기반 이상행위 탐지 기술을 이용해 디바이스 유효성 검증 기술을 제안하고 성능 평가를 진행하여 SDP 환경의 디바이스 유효성 검증 기술을 수행할 수 있는 방안을 제시한다.

The pandemic has rapidly developed a non-face-to-face environment. However, the sudden transition to a non-face-to-face environment has led to new security issues in various areas. One of the new security issues is the security threat of insiders, and the zero trust security model is drawing attention again as a technology to defend against it.. Software Defined Perimeter (SDP) technology consists of various security factors, of which device validation is a technology that can realize zerotrust by monitoring insider usage behavior. But the current SDP specification does not provide a technology that can perform device validation.. Therefore, this paper proposes a device validation technology using SVDD-based abnormal behavior detection technology through user behavior monitoring in an SDP environment and presents a way to perform the device validation technology in the SDP environment by conducting performance evaluation.

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.

생물전환에 의한 고려엉겅퀴 pectolinarin의 pectolinarigenin 전환 시 지표성분의 함량 및 표준화를 위한 분석법 검증을 실시하였다. 분석법 검증 결과 HPLC를 이용한 분석방법에서 표준용액의 retention time과 발효 고려엉겅퀴 추출물의 retention time이 일치하였으며 동일한 spectrum을 나타내는 것으로 특이성을 확인하였다. Pectolinarin 및 pectolinarigenin의 검량선은 상관계수 값이 모두 1로 나타나 높은 직선성을 보여주어 분석에 적합함을 알 수 있었다. Pectolinarin 및 pectolinarigenin의 검출한계는 각각 $4.25{\mu}g/mL$, $2.46{\mu}g/mL$였고, 정량한계는 $12.88{\mu}g/mL$, $7.46{\mu}g/mL$로 나타났다. Pectolinarin 및 pectolinarigenin의 함량을 확인한 결과 고려엉겅퀴 0시간, 5시간 발효물에서는 pectolinarin만 관찰되었으며, 19시간 발효물에서 생물전환에 의해 pectolinarin이 pectolinarigenin으로 전환되는 것을 확인할 수 있었다. 그 이후 28시간부터 72시간까지의 고려엉겅퀴 발효물에서는 pectolinarigenin만 관찰할 수 있었다. 정밀도 측정 결과 pectolinarin 및 pectolinarigenin은 일간 정밀도에서 각각 0.9%, 0.5%의 정밀도를 보여주었으며 일내 정밀도에서는 각각 0.5%, 0.2%로 높은 정밀성을 나타내었다. 또한 pectolinarin은 99.7~104.0%, pectolinarigenin은 99.7~102.4% 범위의 회수율을 보여주어 실험방법에 대한 정확성을 검증하였다. 본 연구 결과 고려엉겅퀴의 지표성분인 pectolinarin 및 pectolinarigenin의 HPLC를 이용한 동시분석방법이 적합한 분석방법임이 검증되었다.

The aim of this study was to investigate a validation method for determination of pectolinarin and pectolinarigenin in fermented Cirsium setidens Nakai. For validation, the specificity, linearity, precision, accuracy, detection limits, and quantification limits of pectolinarin and pectolinarigenin were measured by HPLC. The results show that the detection limits of pectolinarin and pectolinarigenin were $4.25{\mu}g/mL$ and $2.46{\mu}g/mL$, respectively. The recovery rates of pectolinarin and pectolinarigenin were high in the ranges of 99.7~104.0% and 99.7~102.4%, respectively. Inter-day and intra-day precisions of pectolinarin and pectolinarigenin in fermented Cirsium setidens Nakai were 0.9%, 0.5% and 0.5%, 0.2%, respectively. Therefore, application of pectolinarin and pectolinarigenin was validated by an analytical method as a marker compound in Cirsium setidens Nakai.