• Title/Summary/Keyword: hexane

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Simultaneous analysis of residual glucocorticoids in egg by LC/MS/MS (LC/MS/MS를 이용한 계란 중 잔류 글루코코티코이드의 동시분석)

  • Jang, Mi-Ae;Myung, Seung-Woon
    • Analytical Science and Technology
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    • v.22 no.4
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    • pp.326-335
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    • 2009
  • A specific analytical method able to identify and quantify traces of six glucocorticoids residues in eggs were developed. The extraction and clean-up parameters for simultaneous analysis were evaluated and HPLC and spectrometric conditions were also established. For determination of glucocorticoids, 5 g of egg was transferred into a test tube, adjusted pH 5.2 with acetate buffer and was $\beta$-glucuronidase/arylsulfatase from Helix pomatia added. The mixture was centrifuged and supernatant was extracted twice with 20 mL n-hexane. The extraction was performed with HLB cartridge using methanol, followed by clean-up with silica cartridge using methanol/ethyl acetate (4/6, v/v). The analytes were determined by HPLC/ESI-MS/MS operating in the negative ion mode. Validation studies with fortified egg samples for established method were performed. The result of method validation gave good efficiency, linearity, accuracy and precision. The correlation coefficients ($r^2$) of the calibration curves appeared to be higher than 0.99 in egg, indicating excellent linearity. LOD was ranged 0.09 to $0.17{\mu}g/kg$, and recoveries for most compounds were in the range of 55.7-69.8%. This method can be used to determine ${\mu}g/kg$ levels of glucocorticoids in eggs.

Antimicrobial Activity of Aqueous Ethanol Extracts of Perilla frutescens var. acuta Leaf (차조기의 에탄올과 물 혼합 추출물의 항균활성)

  • Kim, Mi-Hyang;Lee, Nan-Hee;Lee, Myung-Hee;Kwon, Dae-Jun;Choi, Ung-Kyu
    • Journal of the Korean Society of Food Culture
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    • v.22 no.2
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    • pp.266-273
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    • 2007
  • This study was conducted to examine the antimicrobial activity of Perilla frutescens var. acuta leaf fractions extracted with a mixture of ethanol and water. The Ca and Mg contents of the leaf were 595.75 mg% and 467.0 mg%, respectively, and they were the highest among all of the test minerals. The extract yield increased w e content of water in e extraction solvent. Antimicrobial activity against Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa was found in the 50,70 and 95% ethanol extracts. Of the various fractions extracted from the mixture of ethanol and water, the ethyl acetate fraction showed antibacterial activity against all microorganisms tested in this experiment, and the antibacterial activity of ethyl acetate fraction from the water extract was the strongest. The phenol and flavonoid content in the ethyl acetate fraction showed no correlation with the concentration of ethanol in the extract solvent; however, their contents were higher in the 30% ethanol and water extraction which the antimicrobial activity of the extract was the strongest.

Anti-osteoarthritis effect of Boswellia serrata gum resin extract in monosodium iodoacetate-induced osteoarthritic Sprague-Dawley rats (Monosodium iodoacetate 유도 골관절염 동물모델에서 보스웰리아 검레진 추출물의 항골관절염 효과 연구)

  • Jae In Jung;Ryong Kim;Eun Ji Kim
    • Journal of Nutrition and Health
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    • v.56 no.3
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    • pp.231-246
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    • 2023
  • Purpose: The aim of this study was to investigate the anti-osteoarthritic effect of the ethanol extract of Boswellia serrata gum resin (FJH-UBS) enriched with keto-β-boswellic acid and 3-O-acetyl-11-keto-β-boswellic acid compared to the conventional Boswellia serrata extract by adding the process of removing oil with hexane, in the monosodium iodoacetate (MIA)-induced osteoarthritis rat model. Methods: Sprague-Dawley (SD) rats were orally administered 0, 40, or 80 mg of FJH-UBS/kg body weight (BW)/day for 5 weeks and injected with MIA intra-articularly into right knee joints on day 15 to induce osteoarthritis. Changes in the knee joint microarchitecture, cartilage degradation, the expression of inflammatory mediators, cytokines, and matrix metalloproteinases (MMPs) in serum and synovia were observed. Results: Oral administration of FJH-UBS (80 mg/kg BW/day) reduced MIA-induced knee swelling and cartilage degradation and increased the expression of type II collagen and aggrecan in articular cartilage. Furthermore, FJH-UBS administration reduced MIA-induced increases in the serum levels of prostaglandin E2, leukotriene B4, interleukin (IL)-1β, IL-6, and MMP-13, and MIA-induced increases in the mRNA expressions of inducible nitric oxide synthase, cyclooxygenase-2, 5-lipoxygenase, IL-1β, IL-6, TNF-α, MMP-2, MMP-9, and MMP-13 in the synovia of knee joints. Conclusion: These results indicate that FJH-UBS exerts its anti-osteoarthritic effects by suppressing the expressions of inflammatory cytokines and MMPs and, thus, cartilage degradation. Furthermore, they suggest that FJH-UBS has potential use as a functional food that improves joint and cartilage health.

Evaluation of radical scavenging and diasestive enzyme inhibitory capacities of peach twigs fraction extract (Prunus persica L. Bastch) (복숭아 나뭇가지 분획추출물의 라디칼 소거 및 소화효소 저해활성 평가)

  • Youjeoung Lee;Gyeong Han Jeong;Ju Yeon Hong;Tae Hoon Kim
    • Food Science and Preservation
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    • v.30 no.1
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    • pp.170-178
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    • 2023
  • We investigated the free radical scavenging and digestive enzyme inhibitory activities of the hot water extract of peach twig (Prunus persica L. Bastch). This extract of the peach twigs was further split up into n-hexane, ethyl acetate (EtOAc), and n-butyl alcohol(n-BuOH), which resulted in three solvent-soluble fractions. Free radical scavenging activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) assay systems, while hypoglycemic effect of the peach twig extract and the solvent-soluble fractions were tested using α-glucosidase and α-amylase inhibition assays. Accordingly, the EtOAc layer showed a greater free radical scavenging activity compared to other solvent-soluble fractions. Furthermore, based on the α-glucosidase and α-amylase assays, the IC50 values were determined to be 38.2±1.6 and 69.6±6.1 ㎍/mL for the EtOAc-soluble fractions, respectively. Taken together, these results suggest that the fractions obtained from the peach twig extract can be considered as a potential source of natural antioxidant and hypoglycaemic constituents.

Anti-oxidant activities of ethanol extract and fractions from defatted Camellia japonica L. seeds (동백 유박 에탄올추출물 및 분획물의 항산화 활성)

  • Weon Pho Park;Nan Kyung Kim;Seok Hee Han;Sanghyun Lee;Ji Hyun Kim;Jine Shang Choi
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.503-511
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    • 2023
  • The aim of this study was to investigate in vitro antioxidant activities of defatted Camellia japonica L. seeds (DCJS). The DCJS were extracted using ethanol and then fractionated with butanol (BuOH), ethyl acetate (EtOAc), chloroform, and hexane. To evaluate antioxidant activity of extract and fractions from DCJS, we investigated free radical scavenging activities such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+), hydroxyl radical (OH), and superoxide anion (O2-) radicals. The five extract and fractions of DCJS dose-dependently increased DPPH, ABTS+ and O2- radical scavenging activities. The BuOH fraction of DCJS showed the highest free radical scavenging activities among other extract and fractions. The contents of total polyphenol and flavonoid in BuOH fraction of DCJS were 23.26 mg GAE/g and 32.39 mg QE/g, respectively. The polyphenol and flavonoids contents of BuOH fraction has highest than other extract and fractions. In addition, BuOH and EtOAc fraction of DCJS contained 102.37 and 165.05 ㎍/g of camelliaside B, respectively. Therefore, DCJS has higher antioxidant activity and may be useful as a natural antioxidant material.

Inhibitory Effect of Chloroform Extract of Marine Algae Hizikia Fusifomis on Angiogenesis (Hizikia fusiformis 클로로포름 추출물의 in vitro 및 in vivo 혈관신생 억제 연구)

  • Myeong-Eun Jegal;Yu-Seon Han;Shi-Young Park;Ji-Hyeok Lee;Eui-Yeun Yi;Yung-Jin Kim
    • Journal of Life Science
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    • v.34 no.6
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    • pp.399-407
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    • 2024
  • Angiogenesis is the process by which new blood vessels form from existing blood vessels. This phenomenon occurs during growth, healing, and menstrual cycle changes. Angiogenesis is a complex and multifaceted process that is important for the continued growth of primary tumors, metastasis promotion, the support of metastatic tumors, and cancer progression. Impaired angiogenesis can lead to cancer, autoimmune diseases, rheumatoid arthritis, cardiovascular disease, and delayed wound healing. Currently, there are only a handful of effective antiangiogenic drugs. Recent studies have shown that natural marine products exhibit antiangiogenic effects. In a previous study, we reported that the hexane extract of H. fusiformis (HFH) could inhibit the development of new blood vessels both in vitro and in vivo. The aim of this study was to describe the inhibitory effect of chloroform extracts of H. fusiformis on angiogenesis. To investigate how chloroform extract prevents blood vessel growth, we examined its effects on HUVEC, including cell migration, invasion, and tube formation. In a mouse Matrigel plug assay, H. fusiformis chloroform extract (HFC) also inhibited angiogenesis in vivo. Certain proteins associated with blood vessel growth were reduced after HFC treatment. These proteins include vascular endothelial growth factor (VEGF), mitogen-activated protein kinase (MAPK)/extracellular signal transduction kinase, and serine/threonine kinase 1 (AKT). These studies have shown that the chloroform extract of H. fusiformis can inhibit blood vessel growth both in vitro and in vivo.

Time Course Variation of Vitamin $C_3$ Content in Leg Skin of Broiler Chicks Exposed to Different Dose of UVB Light (자외선의 상이한 선양을 조사한 브로일러 병아리의 다리 피부중 비타민 $C_3$ 함양의 경시적 변화)

  • 장윤환;김강수;여영수;강훈석;조인호;배은경
    • Korean Journal of Poultry Science
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    • v.20 no.2
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    • pp.93-105
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    • 1993
  • This study was carried out to determine the concentrations of previtamin D$_3$(PreD$_3$), lumisterol$_3$(L3), tachystero1$_3$(73), vitamin D$_3$(VD$_3$) and provitamin D$_3$(ProD$_3$) in leg skins of broiler chicks exposed to UVB lights (maximum intensity at 297 nm) with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min irradiation) . The broiler Hubbard line day old chicks(2 dose $\times$9 elapsed time $\times$4 replica+10 control=82) were fed VD-deficient diet for 31 days in a windowless subdued light room. The skin was collected at 0, 6, 12, 18, 30, 42, 66, 90 or 138 hr after UVB irradiation. The skin lipid was extracted by 9% ethyl acetate/n-hexane, and the fraction of VD$_3$ and its analogues was purified by Sep-Pak silica cartridge. The straight phase HPLC was utilized to analyze ProD$_3$ and its products. The mole %(absolute level expressed in ng/$\textrm{cm}^2$) of PreD$_3$ in leg skin (epidermis+dermis) was 4.67%(44 ng/$\textrm{cm}^2$) or 3.97%(37 ng/$\textrm{cm}^2$) right after UVB irradiation by 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min) at 15 cm distance, respectively. It content in leg skin at 0 hr after exposure was 7.24%(12 ng/$\textrm{cm}^2$) or 0.92%(9 ng/$\textrm{cm}^2$), respectively. The increase in irradiation dose did not affect proportionally the If synthesis.73 concentration in leg skin was 0.58%(S ng/$\textrm{cm}^2$) or 0.57%(6 ng/$\textrm{cm}^2$), respectively 0 hr after irradiation. The VD$_3$ in leg skin of birds exposed to UVB light with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$ was 2.13% (21 ng/$\textrm{cm}^2$) or 0.97% (16ng/$\textrm{cm}^2$), respectively at 0 hr after exposure, 2.72%(26ng/$\textrm{cm}^2$) or 3.84%(37ng/$\textrm{cm}^2$), respectively at 6 hr, and 4.30% ((33ng/$\textrm{cm}^2$) or 6.40%(76ng/$\textrm{cm}^2$), respectively at 12 hr. The peak concentration of VD$_3$ was presented at 18 or 30 hr when 0.204 or 0.409 mJ/$\textrm{cm}^2$) was treated, respectively. It was shown that 18~30 hr were necessary for the thermal conversion of PreD$_3$ into VD$_3$ in the leg skin of broiler chicks. The ProD$_3$ contents in leg skins of negative control, 0.204 mJ/$\textrm{cm}^2$ and 0.409 mJ/$\textrm{cm}^2$ treated birds were 966, 948 and 815 ng/$\textrm{cm}^2$, respectively at right before and after UVB exposure. It was estimated that 18 or 151 ng/$\textrm{cm}^2$ of ProD$_3$ was isomerized to PreD$_3$, L$_3$, T$_3$ and VD$_3$ when exposed to 0.204 or 0.409 mJ/$\textrm{cm}^2$, respective)y. Consequently it was shown that when double dose of UVB light was applied to irradiate the chick body, more but not double synthesis of VD$_3$ and its analogues was occured in leg skin of brolier chicks.

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Antioxidant and α-glucosidase inhibitory effects of ethanolic extract of Ainsliaea acerifolia and organic solvent-soluble fractions (단풍취 추출물 및 분획물의 항산화 및 α-glucosidase 저해 활성 평가)

  • Lee, Eun-Woo;Kim, Taewan;Kim, Hyun-Seok;Park, Youn-Moon;Kim, Seong-Ho;Im, Moo-Hyeog;Kwak, Jae Hoon;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.22 no.2
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    • pp.275-280
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    • 2015
  • Among the naturally occurring antioxidants, polyphenols are widely distributed in various fruits, vegetables, wines, juices, and plant-based dietary sources and divided into several subclasses that included phenolic acid, flavonoids, stilbenes, and lignans. As part of our continuing search for bioactive food ingredients, the antioxidant and ${\alpha}$-glucosidase inhibitory activities of the aqueous ethanolic extract from the aerial parts of Ainsliaea acerifolia were investigated in vitro. The antioxidant properties were evaluated via radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals. In addition, the anti-diabetic effect of A. acerifolia extracts was tested via ${\alpha}$-glucosidase inhibitory assay. Furthermore, the total phenolic contents were determined using a spectrophotometric method. All the tested samples showed dose-dependent radical scavenging and ${\alpha}$-glucosidase inhibitory activities. In particularly, the ${\alpha}$-glucosidase inhibitory and radical scavenging properties of the ethyl acetate (EtOAc)-soluble portion from the aerial parts of the A. acerifolia were higher than those of the other solvent-soluble portions. These results suggest that A. acerifolia could be considered a new potential source of natural antioxidants and antidiabetic ingredients. More systematic investigation of the aerial parts of A. acerifolia will be performed for the further development of anti-oxidative and antidiabetic drugs.

Phytochemical Analysis and Anti-cancer Investigation of Boswellia Serrata Bioactive Constituents In Vitro

  • Ahmed, Hanaa H;Abd-Rabou, Ahmed A;Hassan, Amal Z;Kotob, Soheir E
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7179-7188
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    • 2015
  • Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.

Isolation of Isoflavones and Soyasaponins from the Germ of Soybean (콩 배아로 부터 Isoflavone과 Soyasaponin의 동시 분리)

  • Kim, Sun-Lim;Lee, Jae-Eun;Kim, Yul-Ho;Jung, Gun-Ho;Kim, Dea-Wook;Lee, Choon-Ki;Kim, Mi-Jung;Kim, Jung-Tae;Lee, Yu-Young;Hwang, Tae-Young;Lee, Kwang-Sik;Kim, Wook-Han;Kwon, Young-Up;Kim, Hong-Sig;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.2
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    • pp.149-160
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    • 2013
  • The objective of present study was to simultaneously isolate of isoflavone and soyasaponin compounds from the germ of soybean seeds. Soy germ flours were defatted with hexane for 48h at room temperature, and methanolic extracts were prepared using reflux apparatus at $90^{\circ}C$ for 6h, two times. After extraction, extracts were separated with preparative RP-$C_{18}$ packing column ($125{\AA}$, $55-105{\mu}m$, $40{\times}150mm$), and collected 52 fractions were identified with TLC plate (Kieselgel 60 F-254) and HPLC, respectively. Among the identified isoflavone and soyasaponin fractions, isoflavone fractions were re-separated using a recycling HPLC with gel permeation column (Jaigel-W252, $20{\times}500mm$). Final fractions were air-dried, and the purified compounds of two isoflavones (ISF-1-1, ISF-1-2) and four soyasaponins (SAP-1, SAP-2, SAP-3, SAP-4) were obtained. Two isoflavone compounds (ISF-1-1, ISF-1-2) were acid-hydrolyzed for the identification of their aglycones, and confirmed by comparing with 12 types of isoflavone isomers. While the four kinds of soyasaponins were identified by using a micro Q-TOF mass spectrometer in the ESI positive mode with capillary voltage of 4.5kV, and dry temperature of $200^{\circ}C$. Base on the obtained results, it was conclude that ISF-1-1 is the mixture isomers of daidzin (43.4%), glycitin (47.0%), and genistin (9.6%), but ISF-1-2 is the single compound of genistin (99.8% <). On the other hand, soyasaponin SAP-1 is the mixture compounds of soyasaponin A-group (Aa, Ab, Ac, Ae, Af); SAP-2 is soyasaponin B-group (Ba, Bb, Bc) and E-group (Bd, Be); SAP-3 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$); SAP-4 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$, ${\beta}a$), respectively.