HPLC/DAD를 이용한 6종(種) 우슬(牛膝)의 분류기준 연구;우슬(牛膝)(쇠무릎, Achyranthes japonica $N_{AKAI}$)로부터 20-hydroxyecdysone 분리.동정 및 산지별 우슬의 HPLC 패턴 비교

김정희,김종문,강대훈,Kim, Jeong-Hi,Kim, Jong-Mun,Kang, Dae-Hoon 대한본초학회 2008 大韓本草學會誌 Vol.23 No.1

Objectives : This study was performed to investigate the discriminative criteria of 6 kinds of Achyranthis Radix by HPLC/DAD. Methods : 20-hydroxyecdysone is isolated by silica gel column chromatography ($CHCl_3$:MeOH, 7:1-1:1 v/v) and identified by nuclear magnetic resonance, A high-performance liquid chromatographic method with diode array detection was used to identify 20-hydroxyecdysone in A. japonica. The analysis was performed using $C_{18}$ column with isocratic elution consisted of 18% acetonitrile and 82% water and the detection was carried out by DAD at 254 nm. 6 kinds of Achyranthis Radix from different locations were extracted in MeOH. Each extracts was analyzed by HPLC in same condition as used in analysis of 20-hydroxyecdysone. The identities of each extracts were determined by comparing the retention time and UV spectrum with that of reference compound. Results : 1. A. japonica and A. bidentata showed the similar patterns of HPLC chromatogram and 20-hydroxycedysone was present in both of them because the peaks having the same retention time and UV spectrum as 20-hydroxyecdysone were shown in the HPLC chromatograms of A. japonica and A. bidentata 2. Cyathula officinalis and C. capitata showed the similar patterns of HPLC chromatogram. The peak having the same retention time and UV spectrum as 20-hydroxyecdysone was shown in the HPLC chromatogram of C. capitata but not shown in the HPLC chromatogram of C. officinalis. 3. Two species of medicinal drugs from Sacheon province showed similar patterns of HPLC chromatogram. Achyranthis Radix from Sacheon(wild) did not have 20-hydroxycedysone but Achyranthis Radix from Sacheon(cultivated) showed the peak having the same retention time as 20-hydroxyecdysone but UV spectrum of the peak was different from that of 20-hydroxyecdysone. Conclusions : These results suggested that 20-hydroxyecdysone could be the discriminative criteria for Achyranthis Radix contain 20-hydroxyecdysone though they belong to different genus and species. And the patterns of HPLC chromatogram also could be the discriminative criteria as the different species of Achyranthis Radix belonging to the same genus showed similar patterns of HPLC chromatogram.

PAHs 농도 분포에 따른 GC/MS와 HPLC의 분석특성에 관한 연구

홍좌령 ( Jwa Ryung Hong ),최광민 ( Kwang Min Choi ) 한국산업보건학회 2015 한국산업보건학회지 Vol.25 No.3

Objectives: The purpose of this study was to determine the best method to analyze PAHs at extremely low concentrations. To this end, 16 PAHswere analyzed simultaneously by GC/MS, HPLC/FLD and HPLC/UVD, and the analytical characteristics of HPLC and GC/MS were compared. Methods: This study was conducted by GC/MS and HPLC/FLD/UVD, and evaluated linearity, precision and detection limit. Standard solutions were prepared for 21 samples in the range of 0.00001~1.0 ㎍/mL and the samples were divided into four groups. All samples were made in three sets and analysis was replicated seven times. Results: Sixteen PAHs could be simultaneously separated by HPLC and GC/MS, and the adequate equipment was HPLC/FLD. The retention times by HPLC were shorter than GC/MS, and HPLC had better separation for most PAHs than GC/MS. The peaks of naphthalene and naphthalene-D8 partially overlapped for GC/MS. HPLC/FLD had a 20-2000 times lower limit of detection than GC/MS and UVD. However FLD was not adequate for analyzing acenaphthylene because it has too low a fluorescence quantum yield to be detected. The precision of HPLC/FLD/UVD and GC/MS showed less than 20% at 0.001 ㎍/mL PAHs and when the concentration was higher, the coefficient of variation was decreased. HPLC/FLD was better for the overall detection of limits. Conclusions: The results indicate that the HPLC/FLD method has good linear range, precision and a detection of limits from 0.00001~0.0001 ㎍/mL for all 16 PAHs. This study contributes to providing useful data for analysis technology and can be applied to occupational exposure measurement for PAHs in workplaces.

동백나무 종자의 Camelliaside B 분석을 위한 HPLC-DAD 정량 분석법 개발 및 평가

강주영,윤영대,김봉규 경상국립대학교 농업생명과학연구원 2022 농업생명과학연구 Vol.56 No.6

In this study, we attempted to establish a High-Performance Liquid Chromatography (HPLC) analysis method with Diode Array Detector (DAD) for standard quantification of camelliaside B, which are health functional food ingredient in seed of Camellia japonica. The major compounds of C. japonica seed were identified by MS/MS analysis, comparison retention time, and UV absorbance. HPLC method was validated by specificity, accuracy, precision and limit of quantification. The HPLC method showed that calibration curves for each of the three compounds have high linearity with a correlation coefficient (R2) of > 0.99%. The limits of detection (LOD) and limit of quantitation (LOQ) for camelliaside B were 0.084㎍/㎖ and 0.254㎍/㎖, respectively. Intra-day precision of camelliaside B was 0.09-0.16 RSD. The content of camelliaside B in C. japonica seed extract collected by region by standardized HPLC-DAD analysis varied from 0.279 to 2.05 mg/g. All tested parameters met the appropriate ranges designated in the guideline. Therefore, the HPLC-DAD method established in this study could provide the first step in developing health-related functional products or pharmaceuticals derived from camellia seed extract. 본 연구는 동백나무 종자에 함유된 생리활성물질인 camelliaside B의 표준정량화를 위해 고성능 액체 크로마토그래피 (HPLC-DAD) 분석법을확립하고자 실시하였다. 동백나무 종자에 함유된 주요화합물은 MS/MS 분석, UV 흡광도, 화합물의 유지시간 등을 통해 얻은 결과에 바탕을두어 camelliaside B를 동백나무 종자 추출물의 지표물질로 선정하였다. Camelliaside B의 HPLC 분석법에 대한 특이성, 정확성, 정밀도 및정량한계 등을 검증하였다. Camelliaside B 분석을 위해 표준화된 HPLC 분석법은 0.99% 이상의 상관계수(R2)로 높은 선형성을 가지는 것을보여주었다. Camelliaside B의 회수율은 100.42-108.00%였으며, 검출한계 (LOD)와 정량한계 (LOQ)는 각각 0.084㎍/㎖, 0.254㎍/㎖였다. 동백나무 종자 지표물질 (camelliaside B)의 일중 정밀도는 각각 0.09-0.16 RSD였으며, 일간정밀도는 0.123 RSD였다. 표준화된 분석법으로 지역별로수집한 동백나무 종자의 camelliaside B의 함량은 0.279-2.05㎎/g 으로 다양하였다. HPLC 분석법으로 확인한 모든 매개변수는 기능성 원료인정을 위한 제출자료 작성 기준을 충족하였다. 따라서 본 연구에서 확립한 HPLC 분석법은 동백나무 종자 추출물 유래 건강 관련 기능성 제품또는 의약품 개발의 첫 단계를 제공할 수 있을 것이다.

증기화광산란 검출기를 이용한 콩 함유 수용성 탄수화물의 분석

김경하(Gyeong-Ha Kim),황영선(Young-Sun Hwang),안경근(Kyung-Geun Ahn),김기쁨(Gi-Ppeum Kim),김민지(Min-Ji Kim),홍승범(Seung-Beom Hong),문중경(Jung-Kyeong Moon),정명근(Myoung-Gun Choung) 한국식품영양과학회 2014 한국식품영양과학회지 Vol.43 No.7

In the present study, a new analytical method was devised for the simultaneous determination of soluble carbohydrates in soybean seeds using high performance liquid chromatography/evaporative light scattering detection (HPLC/ELSD). The limit of quantification (LOQ) for soybean soluble carbohydrates ranged from 5.6~7.6 mg/kg using the HPLC/ELSD method and from 16.2~33.9 mg/kg using the high performance liquid chromatography/refractive index detection (HPLC/RID) method. Therefore, the HPLC/ELSD method was more sensitive than HPLC/RID. The precision values for retention time and peak area of the HPLC/ELSD method were evaluated by inter-day (n=5) and intra-day (n=10) assays using a standard solution. All precision values (CV<2.5%) for soybean soluble carbohydrates were acceptable and fulfilled international acceptance criteria. All linear calibration curves were obtained with a correlation coefficient of R<SUP>2</SUP>>0.999. The contents of soluble carbohydrates for the "Shingikong" (yellow soybean) and "Cheongjakong 3" (black soybean) samples were analyzed using the HPLC/RID and HPLC/ELSD methods. The difference in carbohydrate contents between the two detection methods was significant. Carbohydrate contents in the HPLC/ELSD method were higher than those in the HPLC/RID method. Overall, the HPLC/ELSD method showed satisfactory resolution with a favorable LOQ and reproducibility. Therefore, these results indicate that the HPLC/ELSD method may be applied to determine the contents of soluble carbohydrates in soybean seeds and related food stuffs.

GC 및 HPLC 비교분석에 기초한 차조기 종실내 tocotrienol 부재의 평가

이영상,김민경 한국작물학회 2008 한국작물학회지 Vol.53 No.-

Lipid soluble vitamin E consists of tocopherols and tocotrienols depending upon double bonds in phytyl side chains attached to chromanol ring. Recent reports on antioxidative, anticancer, and cholesterol-lowering effects of tocotrienols have increased researches and commercialization of tocotrienols. Purple perilla (Perilla frutescens var. acuta Kudo) has been reported as a plant containing tocotrienols along with tocopherol forms of vitamin E based upon normal phase HPLC analysis. To confirm the existence or absence of tocotrienol form of vitamin E in purple perilla, comparative analysis using HPLC, GC/FID, and GC/MSD has been conducted for 14 purple perilla genetic accessions collected from Korea and Japan. Normal phase HPLC analysis showed α-, β-, γ-, and δ-tocopherols along with peaks with retention times quite similar to β- and γ-tocotrienols. Same purple perilla samples, analysed by GC exhibited α-, β-, γ-, and δ-tocopherols quantitatively equivalent to HPLC results. However, no peaks for β- and γ-tocotrienols could be observed and unknown two peaks of similar retention times with β- and γ-tocotrienols were identified not corresponding tocotrienols by GC/MSD. These results suggest the absence of tocotrienol form of vitamin E in purple perilla as well as the necessity of using GC-based qualitative and quantitative vitamin E analysis to avoid misinterpretation of peaks with similar retention times as tocotrienol isomers when analysed by an HPLC. Lipid soluble vitamin E consists of tocopherols and tocotrienols depending upon double bonds in phytyl side chains attached to chromanol ring. Recent reports on antioxidative, anticancer, and cholesterol-lowering effects of tocotrienols have increased researches and commercialization of tocotrienols. Purple perilla (Perilla frutescens var. acuta Kudo) has been reported as a plant containing tocotrienols along with tocopherol forms of vitamin E based upon normal phase HPLC analysis. To confirm the existence or absence of tocotrienol form of vitamin E in purple perilla, comparative analysis using HPLC, GC/FID, and GC/MSD has been conducted for 14 purple perilla genetic accessions collected from Korea and Japan. Normal phase HPLC analysis showed α-, β-, γ-, and δ-tocopherols along with peaks with retention times quite similar to β- and γ-tocotrienols. Same purple perilla samples, analysed by GC exhibited α-, β-, γ-, and δ-tocopherols quantitatively equivalent to HPLC results. However, no peaks for β- and γ-tocotrienols could be observed and unknown two peaks of similar retention times with β- and γ-tocotrienols were identified not corresponding tocotrienols by GC/MSD. These results suggest the absence of tocotrienol form of vitamin E in purple perilla as well as the necessity of using GC-based qualitative and quantitative vitamin E analysis to avoid misinterpretation of peaks with similar retention times as tocotrienol isomers when analysed by an HPLC.

UPLC를 이용한 밀크씨슬추출물 지표 성분인 실리마린 분석법 검증

유동선 ( Dongsun Yoo ),정경희 ( Kyung Hee Jung ),최승준 ( Seung Jun Choi ) 한국산업식품공학회 2018 산업 식품공학 Vol.22 No.2

밀크씨슬추출물의 지표물질인 실리마린 분석법의 효율성재고를 위하여 건강기능식품공전의 HPLC 분석법과 UPLC를 이용한 신규 분석법을 비교하여 분석법 검증 및 정량적 평가를 실시하였다. UPLC를 이용한 신규 분석법을 검증하기 위하여 특이성, 직선성, 정확성, 정밀성, 실험실 내 정밀성, 완건성 등 6개의 검증항목을 선정하여 UPLC 분석법검증을 실시하였다. 특이성에서 1.5 이상의 분리능을 보여 6가지 실리마린 표준물질이 HPLC 및 UPLC 분석법 모두에서 선택적으로 정확하게 검출됨을 확인하였다. 직선성에서는 25-400 μg/mL의 농도에서 결정계수 값이 0.9999 이상으로 거의 직선에 가까운 값을 보였다. 정확성에서 UPLC를 이용한 신규 분석법의 회수율은 평균 99%이상으로 HPLC 분석법보다 높은 회수율을 보였으며, 회수율편차역시 50% 이상 감소하였다. 정밀성에서도 UPLC 분석법에서 얻어진 피크 면적의 상대표준편차가 HPLC를 이용한 기존 분석법의 절반 이하로 감소하여 검증 항목에서 월등한 결과값을 보였다. 또한 실험실 내 정밀성에서도 UPLC분석법에서 얻어진 상대표준편차가 HPLC 분석법에 얻어진 상대표준편차보다 현저히 낮음을 확인하였다. 분석법검증 결과 6개의 검증항목에서 HPLC와 UPLC 분석법 모두 적합하게 검증되었으며, 검증항목 중 정확성, 정밀성, 실험실 내 정밀성 및 완건성 항목에서 UPLC 분석법의 검증값이 HPLC 분석법보다 뛰어났다. 이상의 결과로 UPLC를 사용한 실리마린의 분석법은 건강기능식품의 실리마린을 분석하는데 충분히 효율적인 분석법이 될 것으로 판단된다. This study attempted to establish an ultra performance liquid chromatography (UPLC) analysis method for standard determination of silymarin as a health functional food material in Silybum marianum extraxt (milk thistle). UPLC was performed on a Waters Acquity BEH C18 (50×2.1 mm, 1.7 μm) column using a gradient elution of distilled water and methanol at a flow rate of 0.21 mL/min and detection wavelength of 288 nm. The UPLC method showed high linearity in the calibration curve at a coefficient of determination (r2) of >0.9999, and limit of detection and quantitation for 6 flavonolignans were 0.0167-0.2469 and 0.1648-1.2931 μg/mL, respectively. The recovery of each flavnolignan was in the range of 99.96-100.81%, and the relative standard deviation for precision of each flavonolignan was less than 1.0%. The UPLC method established in this study was more specific for the quantitative determination of silymarin than the HPLC method. Also, since the UPLC method is shorter in the equipment operation time and smaller in the amount of used solvent than the HPLC method, UPLC is expected to have higher energy efficiency and lower environmental impact compared with HPLC.

Hyphenated-HPLC 기술을 활용한 홍화씨의 항산화 성분 분석

김수진(Su Jin Kim),김상민(Sang Min Kim),강석우(Suk Woo Kang),엄병헌(Byung Hun Um) 한국식품과학회 2010 한국식품과학회지 Vol.42 No.4

본 연구에서는 세 종류의 hyphenated-HPLC 기술을 활용하여 홍화씨로부터 3종의 항산화 화합물의 구조를 규명하였다. 우선 온라인 항산화 분석 장치를 통하여 홍화씨 추출물로부터 ABTS 라디칼 소거활성을 가지는 성분을 검색 및 항산화 정량을 수행한 후, 단일 물질로 분리되고, 항산화 활성을 가지는 세 가지 화합물에 대해서 구조 규명을 시도하였다. 우선 LC-NMR을 이용하여 stop-flow mode에서 이들 세 가지 화합물에 대해 ¹H-NMR spectrum데이터를 얻은 결과 각 화합물은 8’-hydroxyarctigenin-4’-O-β-D-glucoside, N-(p-coumaroyl) serotonin, N-feruloylserotonin으로 확인되었다. 그리고 LC-ESI-MS를 활용하여 각 화합물에 대한 분자량에 대한 정보를 얻어 LC-NMR에서 규명된 화합물이 정확함을 다시 한 번 확인할 수 있었다. 본 연구에서는 기존의 탐색방법인 여러 크로마토그래피 방법이나 preparative HPLC 등을 이용하여 활성물질을 분리하고 off-line NMR, MS 등을 활용하여 구조를 규명하는 방법에 비하여, hyphenated-HPLC 방법을 활용하여 혼합물 상태인 추출물을 분리하지 않고 신속하게 단일성분의 구조를 규명하고, 또한 각각의 성분에 대한 항산화도를 측정할 수 있다는 장점이 있음을 증명하였다. 이는 천연물 또는 식품 분야의 연구에 있어 추출물의 항산화 성분을 분석하고 그 구조를 신속 간편하게 확인할 수 있으므로 항산화 성분 탐색 및 변이 연구에 매우 유용하리라 생각된다. Hyphenated-HPLC techniques combine the separation power of HPLC with the structural and bioactivity information provided by NMR, ESI/MS, and an on-line antioxidant screening system. The major advantages over the traditional off-line techniques are rapidity and efficiency. In this study, we used hyphenated HPLC techniques including online HPLC-ABTS, LC-NMR, and LC-MS todirectly identify the major antioxidants of safflower (Carthamus tinctorius L.) seeds. The results demonstrated that the major antioxidant compounds from on-line HPLC-ABTS analysis were identified as 8’-hydroxyarcgenin-4’-O-β-D-glucoside, N-(p-coumaroyl) serotonin, and N-feruloylserotonin. Among them, N-feruloylserotonin accounted for almost 50% of the ABTS radical scavenging activity of the total extract. The results demonstrate that HPLC hyphenated techniques can be used to rapidly screen and structurally identify antioxidants from crude plant extracts.

GC 및 HPLC 비교분석에 기초한 차조기 종실내 tocotrienol 부재의 평가

이영상,김민경 韓國作物學會 2008 한국작물학회지 Vol.53 No.5

비타민 vitamin E 구성 성분으로 항산화, 항암, 고지혈증 개선 등 다양한 생리활성을 나타내는 tocotrienol이 국내산 차조기에서 보고된 바, 존재 여부 확인을 위하여 국내수집 6종, 일본수집 8종 등 총 14종의 차조기 유전자원과 들깨 종실을 대상으로 HPLC 및 GC, GC/MS 비교 분석을 수행 하였으며 그 결과는 다음과 같다. 1. 순상의 HPLC 이용시 차조기에서 4종의 tocopherol 및β-tocotrienol , λ-tocotrienol 과 매우 유사한 머무름 시간의 발견되었다. 2. 그러나 동일 시료를 GC로 재분석한 결과 13개 차조기 유전자원에서 모두 4종의 tocotrienol과 일치하는 물질은 발견되지 않았다. 3. GC 이용시 δ-tocotrienol 및 λ-tocotrienol 과 유사한 머무름 시간의 peak가 검출되었으나 GC/MS를 이용하여 확인한 결과 tocotrienol이 아닌 것으로 확인되었다. 4. 이상의 HPLC 및 GC 비교분석결과에 기초할 때, 차조기에는 tocotrienol류의 vitamin E는 존재하지 않는 것으로 추정되었으며 GC를 이용한 vitamin E isomer 분석이 HPLC 이용시 발생가능한 유사 머무름시간대 물질을 tocotrienol로 판단하는 오류 방지에 효과적임을 알 수 있었다. Lipid soluble vitamin E consists of tocopherols and tocotrienols depending upon double bonds in phytyl side chains attached to chromanol ring. Recent reports on antioxidative, anticancer, and cholesterol-lowering effects of tocotrienols have increased researches and commercialization of tocotrienols. Purple perilla (Perilla frutescens var. acuta Kudo) has been reported as a plant containing tocotrienols along with tocopherol forms of vitamin E based upon normal phase HPLC analysis. To confirm the existence or absence of tocotrienol form of vitamin E in purple perilla, comparative analysis using HPLC, GC/FID, and GC/MSD has been conducted for 14 purple perilla genetic accessions collected from Korea and Japan. Normal phase HPLC analysis showed α- , β- , ~gamma- , and δ-tocopherols along with peaks with retention times quite similar to β- and ~gamma-tocotrienols . Same purple perilla samples, analysed by GC exhibited α- , β- , ~gamma- , and δ-tocopherols quantitatively equivalent to HPLC results. However, no peaks for β- and ~gamma-tocotrienols could be observed and unknown two peaks of similar retention times with β- and ~gamma-tocotrienols were identified not corresponding tocotrienols by GC/MSD. These results suggest the absence of tocotrienol form of vitamin E in purple perilla as well as the necessity of using GC-based qualitative and quantitative vitamin E analysis to avoid misinterpretation of peaks with similar retention times as tocotrienol isomers when analysed by an HPLC.

Simultaneous determination of chromones and coumarins in Radix Saposhnikoviae by high performance liquid chromatography with diode array and tandem mass detectors

Kim, Min Kyung,Yang, Dong-Hyug,Jung, Mihye,Jung, Eun Ha,Eom, Han Young,Suh, Joon Hyuk,Min, Jung Won,Kim, Unyong,Min, Hyeyoung,Kim, Jinwoong,Han, Sang Beom Elsevier 2011 Journal of chromatography A Vol.1218 No.37

<P><B>Abstract</B></P><P>Methods using high performance liquid chromatography with diode array detection (HPLC-DAD) and tandem mass spectrometry (HPLC–MS/MS) were developed and validated for the simultaneous determination of 5 chromones and 6 coumarins: prim-<I>O</I>-glucosylcimifugin (1), cimifugin (2), nodakenin (3), 4′-<I>O</I>-β-<SMALL>D</SMALL>-glucosyl-5-<I>O</I>-methylvisamminol (4), sec-<I>O</I>-glucosylhamaudol (5), psoralen (6), bergapten (7), imperatorin (8), phellopterin (9), 3′-<I>O</I>-angeloylhamaudol (10) and anomalin (11), in Radix Saposhnikoviae. The separation conditions for HPLC-DAD were optimized using an Ascentis Express C18 (4.6mm×100mm, 2.7μm particle size) fused-core column. The mobile phase was composed of 10% aqueous acetonitrile (A) and 90% acetonitrile (B) and the elution was performed under a gradient mode at a flow rate of 1.0mL/min. The detection wavelength was set at 300nm. The HPLC-DAD method yielded a base line separation of the 11 components in 50% methanol extract of Radix Saposhnikoviae with no interfering peaks detected. The HPLC-DAD method was validated in terms of linearity, accuracy and precision (intra- and inter-day), limit of quantification (LOQ), recovery, and robustness. Specific determination of the 11 components was also accomplished by a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization (ESI) source. This HPLC–MS/MS method was also validated by determining the linearity, limit of quantification, accuracy, and precision. Quantification of the 11 components in 51 commercial Radix Saposhnikoviae samples was successfully performed using the developed HPLC-DAD method. The identity, batch-to-batch consistency, and authenticity of Radix Saposhnikoviae were successfully monitored by the proposed HPLC-DAD and HPLC–MS/MS methods.</P>

다양한 낙농 발효유제품에서 HPLC를 이용하여 탄수화물과 유기산의 동시 검출에 관한 연구

김동현 ( Dong Hyeon Kim ),황대근 ( Dae Geun Hwang ),천정환 ( Jung Whan Chon ),김현숙 ( Hyunsook Kim ),김홍석 ( Hong Seok Kim ),송광영 ( Kwang Young Song ),임진혁 ( Jin Hyuk Yim ),김영지 ( Young Ji Kim ),강일병 ( Il Byung Kang ) 한국유가공기술과학회 2015 Journal of Dairy Science and Biotechnology (JMSB) Vol.33 No.4

본 연구에서는 다양한 낙농 발효유제품에 있어서 다양한 유기산과 탄수화물의 분석이 HPLC 방법으로 동시에 잘 진행되었다. 이 방법의 주요 장점으로는 (1) 간단한 시료 준비와 단일 추출이 가능, (2) 유동상(mobile phase)과 추출 용매에 있어서 편리성과 경제성, (3) 추출제와 유동상에 사용되는 황산의 농도가 매우 낮기에 친환경적인 실험, (4) 하나의 샘플당 30분 안에 HPLC의 분석이 완료 가능, (5) 다양한 농도 범위에서도 분석 및 정량 가능, (6) 몇 가지 물질을 제외하고 모든 화합물의 변동계수가 낮음, 그리고 (7) 회수율이 대체적으로 높다는 것이다. 또한, 60~65℃에서 분석 column을 사용하였기에 column 효율면에서도 손실이 극히 적은 것으로 관찰되었다. 따라서 본 실험에서 분석된 다양한 낙농 발효유제품에서의 acid, 탄수화물, diacetyl, acetoin 등이 HPLC 방법은 매우 신속하고 정확하게 검출되었다. 하지만 다양한 향미의 변화를 최소화하면서 다양한 향미를 효과적으로 추출할 수 있는 HPLC 방법들이 지속적으로 개발되어야 다양한 낙농 발효유제품의 품질개선에 기여할 것으로 사료된다. 왜냐하면 향미 성분의 분석은 성분을 정성하는 것뿐만 아니라, 정성된 화합물이 그 식품의 향에 어느 정도 기여하는지를 중요하게 결정하기 때문이다. 또한 본 실험에 사용된 HPLC 방법으로 다양한 유산균주의 일정한 배양시간동안 유당분해효소(lactase) 능력이 조사 되었는데, 각 Probiotic lactic acid bacteria마다 다양한 유당분해효소(lactase)의 능력을 보였으며, 같은 균종에서도 차이가 있는 것으로 나타났다. 본 실험에서는 Bifidobacterium spp.가 Lactobacillus spp.와 Streptococcus spp.보다 높은 유당분해효소(lactase)의 능력을 보였다. 하지만 유산균주의 유당분해효소 능력을 정확하게 파악하기 위해서는 각 Probiotic lactic acid bacteria의 exo- 및 endo-lactase에 대한 연구가 추가적으로 필요하다. 결론적으로 다양한 낙농발효식품 및 기타 발효식품 등에서 HPLC 방법으로 동시에 유기산과 탄수화물 분석이 공인방법으로써 인정되기 위해서는 더 많은 기초자료 확립과 재현성에 관한 지속적인 연구가 진행되어야 할 것으로 사료된다. Various carbohydrates (lactose, glucose, and fructose), lactic acid, uric acid, and acetoin were separated on a ZORBAX Carbohydrate Analysis column using the Agilent 1200 HPLC ChemStationTM, and were identified according to retention times with 325 Dual Wavelength UV-Vis Detector and Refractive Index Detector with 0.013 N H2SO4 at a flow rate of 0.8 mL/min. In addition, the lactase activity of four commercial probiotic lactic acid bacteria during 6-hour incubation was determined using a high-performance liquid chromatography (HPLC) method. Among the tested samples, Bifidobacterium animalis subsp. lactis showed the greatest lactase activity, followed by Lactobacillus rhamnosus and Lactobacillus casei, with Streptococcus salivarius subsp. thermophilus showing the lowest activity. Therefore, this HPLC technique shows potential for evaluating the fermentation processes of probiotic lactic acid bacteria and could simultaneously confirm the degree of ripening in various fermented dairy foods within only a half hour.