• The Korean Journal of Zoology
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• v.33 no.1
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• pp.35-44
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• 1990

Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.565-573
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• 1998

To study the specific tools for the diagnosis of Newcastle disease virus (NDV) in chicken, polymerase chain reaction (PCR) and its presumable conditions were evaluated for the detection of hemagglutinin-neuraminidase (HN) gene of NDV RNA. For these purposes, Kyojeongwon strain of the NDV was propagated in allantoic cavity of SPF embryonating chicken eggs, and viral RNA was extracted from fractionated virus after the allantoic fluids were ultracentrifuged with sucrose gradient. The first-strand cDNA was then made for the HN gene of NDV RNA by reverse transcription at $42^{\circ}C$ for 1 hour using specific primer complementary to the HN gene. The single-stranded cDNA was used as template in the PCR of the HN-DNA, and various conditions of the PCR were evaluated to set up method for the specific detection of the HN-DNA. The PCR conditions promising for the detection of HN gene consist of preheating at $94^{\circ}C$, 5 min, 30 cycles of denaturation at $94^{\circ}C$, 1 min, annealing at $55^{\circ}C$, 1 min and polymerization at $72^{\circ}C$, 2 min, and a cycle of extension at $72^{\circ}C$, 5 min. when NDVs of allantoic fluids without fractionation were applied to the above PCR condition, the HN genes were detected effectively not only from Kyojeongwon but from other velogenic strains such as Herts and a field isolate.

• Analytical Science and Technology
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• v.11 no.6
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• pp.436-439
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• 1998

Ginseng saponins were analysed using reversed-phase high performance liquid chromatography with several columns. The optimum conditions were as following : reverse phase column; Novapak $C_{18}$ ODS column ($3.9mm{\times}150mm$, $5{\mu}m$), acetonitrile/water binary mobile phase gradient controller system, solvent flow rate; 1.5 mL/min, and UV (203 nm) detector. The complete separation of ginsenoside $Rb_1$, $Rb_2$, Rc, Rd, Re, Rf and $Rg_1$ was achieved within 50 min. The regression coefficients of the calibration curves for seven ginsenosides were 0.98~0.99.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.51-56
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• 2003

The angiotensin I converting enzyme (ACE) has an important role in the maintenance of blood pressure. The ACE inhibitory activities of foods have recently been studied. We tried to isolate ACE inhibitory peptides from the Flavourzyme (FZ), Pescalase (PE), and Thermolysine (TH) protease digests of corn gluten, which was restricted to the use the source of food for digestion problem. The FZ, PE, TH/PE protease hydrolyzed corn gluten and the inhibitory activities of the hydrolyzates for ACE were measured. Major fractions were isolated from the digests using ODS chromatography after treating with ethanol in step gradient. The ACE inhibitors were further purified by Bio-Gel P-2 column and reverse phase HPLC. Five inhibitory peptides were isolated. Their amino acids were sequenced as LPF ($IC_{50}$ = 40$\mu$M), GPP ($IC_{50}$ = 17.6$\mu$M), PNPY ($IC_{50}$ = 30.7$\mu$M), SPPPFYL ($IC_{50}$ = 63 $\mu$M), and SQPP ($IC_{50}$ = 17.2$\mu$M).

• Journal of Ginseng Research
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• v.44 no.4
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• pp.563-569
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• 2020

Background: White ginseng consists of the roots and rhizomes of the Panax species, and red ginseng is made by steaming and drying white ginseng. While red ginseng has both polar and nonpolar ginsenosides, previous studies showed white ginseng to have only polar ginsenosides. Because nonpolar ginsenosides are formed through the manufacture of red ginseng from white ginseng, researchers have generally thought that nonpolar ginsenosides do not exist in white ginseng. Methods: We developed a simultaneous quantitative method for six nonpolar ginsenosides in white ginseng using reverse-phase high-performance liquid chromatography coupled with integrated pulsed amperometric detection. The nonpolar ginsenosides of white ginseng were extracted for 4 h under reflux with 50% methanol. Results: Using the gradient elution system, all target components were completely separated within 50 min. Nonpolar ginsenosides were determined in the rhizome head (RH), main root (MR), lateral root, and hairy root (HR) of 6-year-old white ginseng samples obtained from several regions (Geumsan, Punggi, and Kanghwa). The total content in the HR of white ginseng was 37.8-56.8% of that in the HR of red ginseng. The total content in the MR of white ginseng was 5.9-24.3% of that in the MR of red ginseng. In addition, the total content in the RH of white ginseng was 28.5-35.8% of that in the HR of red ginseng Conclusion: It was confirmed that nonpolar ginsenosides known to be specific components of red ginseng were present at substantial concentrations in the HR or RH of white ginseng.

• Advances in Traditional Medicine
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• v.4 no.4
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• pp.267-273
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• 2004

Oxidative stress is associated with many kinds of chronic diseases. Antioxidants such as polyphenols are compounds that protect cells against the damaging effects of reactive oxygen species. Grape seeds are considered good resources of polyphenols, and grape seed extracts have a very strong antioxidant effect. In the present study, we established a simple gradient reverse-phase high performance liquid chromatography method to determine polyphenol content from three different grape seed resources. An ODS (2), $150\;{\times}\;3.2\;mm$ column has been employed, and six polyphenols have been determined: gallic acid, protochatechuic acid, (+)-catechin, (-)-epicatechin, procyanidin B2, and epicatechin gallate. Catechin and epicatechin were the main polyphenol compounds in all three extracts. The amount of procyanidin B2 was higher in Extract 1 (from a company of China), while Extract 2 (extracted in our lab) and Extract 3 (from a company of USA) contained higher proportions of epicatechin gallate. For the total polyphenol content, Extract 1 was much higher than that of Extract 2 and 3. The results suggest that the dietary dose of grape seed extracts from different resources should be adjusted according to polyphenol content.

• Food Science and Biotechnology
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• v.15 no.5
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• pp.778-780
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• 2006

High performance liquid chromatography (HPLC) was used for the analysis of the lignans eleutheroside B and eleutheroside E in the fruits of Acanthopanax species. A reverse-phase system using a gradient of $H_2O$ and acetonitrile as the mobile phase was developed and detection was at 210 nm. The analysis was successfully carried out within 20 min. The content of eleutheroside B and eleutheroside E in Acanthopanax species was measured in the fruits of A. senticosus (0.58 and $1.66\;{\mu}g/mg$, respectively), A. sessiliflorus (1.15 and $8.49\;{\mu}g/mg$, respectively), A. koreanum (2.16 and $1.80\;{\mu}g/mg$, respectively), and A. divaricatus (1.06 and $7.08\;{\mu}g/mg$, respectively).

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1459-1464
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• 2004

The purpose of this research was to investigate the potential use of cheese whey protein (CWP), a cheese by-product. The physiological activity of calcium-binding peptides in CWP may be used as a food additive that prevents bone disorders. This research also examined the characteristics of calcium-binding peptides. After the CWP was heat treated, it was hydrolyzed by trypsin. Then calcium-binding peptides were separated and purified by ion-exchange chromatography and reverse phase HPLC, respectively. To examine the characteristics of the purified calcium-binding peptides, amino acid composition and amino acid sequence were analyzed. Calcium-binding peptides with a small molecular weight of about 1.4 to 3.4 kDa were identified in the fraction that was flowed out from 0.25 M NaCl step gradient by ion-exchange chromatography of tryptic hydrolysates. The results of the amino acid analysis revealed that glutamic acid in a calcium-binding site took up most part of the amino acids including a quantity of proline, leucine and lysine. The amino acid sequence of calcium-binding peptides showed Phe-Leu-Asp-Asp-Asp-Leu-Thr-Asp and Ile-Leu-Asp-Lys from $\alpha$-LA and Ile-Pro-Ala-Val-Phe-Lys and Val-Tyr-Val-Glu-Glu-Leu-Lys from ${\beta}$-LG.

• Korean Journal of Agricultural Science
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• v.38 no.2
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• pp.295-299
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• 2011

The content of hesperidin in the mixed tea, which was composed of dried orange peel, laurel leaf, mulberry leaf, silver magnolia leaf, oriental melon tap, cassia seed, and licorice root, was determined by high performance liquid chromatography (HPLC). Hesperidin was quantified by a reverse phase column with gradient solvent system (watcr:acetonitrile = 80:20 to 35:65 for 30 min) and UV/VIS detection (280 nm). The How rate was kept constant at 1.0 ml/min. The content of hesperidin in the mixed tea was measured in depending on extraction time 1, 2, 3, and 4 min (29.07, 52.39, 52.45, and 88.35 mg/g, respectively).

• YAKHAK HOEJI
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• v.50 no.1
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• pp.21-25
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• 2006

Reiterated normal-phase column chromatography lead to the isolation and purification of six known compounds but for the first time from the whole plant of Sasa borealis (Hack.) Makino (Gramineae): tricin 4'-O-(erythro-${\beta}$-guaia-cylglyceryl) ether (1), tricin 4'-O-(threo-${\beta}$-guaiacylglyceryl) ether (2), tricin 4'-O-[erythro-${\beta}$-guaiacyl-(9'-O-acetyl)-glyceryl] ether (3), tricin 4'-O-[threo-${\beta}$-guaiacyl-(9'-O-acetyl)-glyceryl] ether (4), (-)-pinoresinol (5), and vanillin (6). The structures of the compounds (1-6) were established based on interpretation of high resolution NMR (COSY, HSQC, HMBC, and NOESY) spectral data. In particular, compounds 1 and 3 were diastereomers of compounds 2 and 4, respectively. These two sets of diastereomers were able to be simultaneously identified and quantified by a gradient reversed-phase HPLC method with UV photodiode array, This sensitive HPLC method is noteworthy as a simultaneous separation and identification method to test the extract of the family Gramineae which contains these compounds.