• Title/Summary/Keyword: HPLC analysis method

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Analysis of Marker Substances in Samul-tang by HPLC-MS/MS (HPLC-MS/MS에 의한 사물탕의 지표성분 분석)

  • Yu, Young-Beob;Kim, Mi-Jung;Huang, Dae-Sun;Ha, Hye-Kyeong;Ma, Jin-Yeul;Shin, Hyeun-Kyoo
    • The Korea Journal of Herbology
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    • v.22 no.2
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    • pp.97-102
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    • 2007
  • Objectives : This study presents a high performance liquid chromatography - electrospray ionization-mass spectrometer (HPLC-MS/MS) methods for the quantitative and qualitative analysis of various active components in Samul-tang, which is composed of four crude herbs. Methods : HPLC-ESI-MS/MS for the determinations of paeoniflorin and 5-HMF (5-hydroxymethyl 2-furaldehyde) in the Samul-tang, the separation method was performed on an COSMOS1L 5C18-AR-Il (2.0 X 150 mm I.D.) column by gradient elution with 0.1% acetic acid and 5% CH3CN in water (A)-0.1% acetic acid and 5% H20 in CH3CN (B) as the mobile phase at a flow-rate of 300 ${\mu}L/min$ with detection at quadrupole mass spectrometer. The all marker substances were always detected as the base peaks in the positive ion mode. Results : The paeoniflorin of Paeoniae Radix in Samul-tang showed a strong base peak [M+H2O]+ in the positive detection mode to give the following as; paeoniflorin (498.109 [M+H2O]+, 479.8 [M]+, 301 [M-glucose]+, 179.3 [glucose]+). Based on the HPLC retnetion time and MS of standard compounds confirmed the identity of active compounds in Rehmanniae Radix Preparata as follows; 5-HMF (127.0[M+H]+, 109.3 [M-OH]+) in the positive ion mode. Conclusion : According to the above results, HPLC-ESI-MS method permits assignment of tentative structures such as paeoniflorin and 5-HMF in the Samul-tang.

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Quality Evaluation of Herbal Prescription, Oc Chun San, Employing Simultaneous Determination of the Marker Compounds by HPLC (HPLC를 이용한 옥천산 중 갈근, 감초, 오미자 지표성분의 다성분 동시분석)

  • Yoo, Jeong-Lim;Jang, Dae-Sik;Kim, Jin-Sook
    • Korean Journal of Oriental Medicine
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    • v.11 no.2
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    • pp.167-178
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    • 2005
  • As a part of the quality control of herbal prescriptions which has been used for diabetes and related diseases, a reversed-phase liquid chromatographic method was developed for the simultaneous quantification of the three marker compounds, puerarin (Puerariae Radix), glycyrrhizin (Glycyrrhizae Radix), schizandrin (Schizandrae Fructus) in Oc Chun San. The HPLC analysis method was validated for parameters such as linearity, Limits of Detection(LOD), quantification(LOQ), repeatability, stability and recovery.

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Determination of Vitamin $B_{12}$ in Foods Using Column-Switching Technique in $\mu$-HPLC ($\mu$-HPLC의 Column-Switching 기술을 이용한 식품중 비타민 $B_{12}$의 분석)

  • 박성진;김혜경;함태식;김병용
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.6
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    • pp.1208-1211
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    • 1999
  • Semi HPLC using a column switching technique was used to determine the trace content of vitamin B12 in various foods. Total analytical time required less than 20 mins per sample and the recovery ratio was 99.9, 99.6, 100.1 and 99.8% for 1.0, 10.0, 100.0 and 1,000 g/kg, respectively. The content of vitamin B12 in various foods obtained using column switching method showed higher levels compared to labels in dried milk(0.5 g/100g) and in grain products(0.51~34.36 g/100g). Thus, this column switching method was more sensitive, effective and precise than the microbiological analysis currently used to determine the trace compounds like a vitamin B12.

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Quantitative Evaluation of Radix Astragali through the Simultaneous Determination of Bioactive Isoflavonoids and Saponins by HPLC/UV and LC-ESI-MS/MS

  • Kim, Jin-Hee;Park, So-Young;Lim, Hyun-Kyun;Park, Ah-Yeon;Kim, Ju-Sun;Kang, Sam-Sik;Youm, Jeong-Rok;Han, Sang-Beom
    • Bulletin of the Korean Chemical Society
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    • v.28 no.7
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    • pp.1187-1194
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    • 2007
  • The three major active isoflavonoids (calycosin-7-O-β -glucoside, isomucronulatol 7-O-β-glucoside, formononetin) and two main saponins (astragaloside I, astragaloside IV) in an extract of Radix Astragali were determined using rapid, sensitive, reliable HPLC/UV and LC-ESI-MS/MS methods. The separation conditions employed for HPLC/UV were optimized using a phenyl-hexyl column (4.6 × 150 mm, 5 μm) with the gradient elution of acetonitrile and water as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 230 nm. The specificity of the peaks was determined using a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization (ESI) source that was operated in multiple reaction monitoring (MRM) in the positive mode. These methods were fully validated with respect to the linearity, accuracy, precision, recovery and robustness. The HPLC/UV method was applied successfully to the quantification of three major isoflavonoids in the extract of Radix Astragali. The results indicate that the established HPLC/UV and LC-ESI-MS/MS methods are suitable for the quantitative analysis and quality control of multi-components in Radix Astragali.

Determination of Flavonoids, Tannins and Ellagic Acid in Leaves from Rubus L. Species

  • Gudej, Jan;Tomczyk, Michal
    • Archives of Pharmacal Research
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    • v.27 no.11
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    • pp.1114-1119
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    • 2004
  • This paper describes the quantitative determination of flavonoids, tannins and ellagic acid in the leaves from wild and cultivated variations of Rubus L. species (Rosaceae): raspberry (2 wild and 13 cultivars) and blackberry (3 wild and 3 cultivars). The content of flavonoids was analyzed using spectrophotometric (the Christ-M llers method) and HPLC analysis after acid hydrolysis. The content of tannins was determined by the weight method, with hide powder, described by German Pharmacopoeia 10 (DAB 10). Ellagic acid content was examined using the HPLC method after acid hydrolysis. Flavonoid content, determined using the Christ-Muller's method was higher for the blackberry leaves than for the raspberry leaves and varied between 0.46% and 1.05%. Quercetin and kaempferol were predominant in all samples analyzed using the HPLC method. The highest flavonoid content was found in the leaves of R. nessensis (1.06%); with results in all of the examined samples varying between 0.27% and 1.06%. The concentration of ellagic acid in all species was determined after acid hydrolysis and ranged from 2.06% to 6.89%. The leaves of raspberries are characterized by greater amounts of tannins (varying between 2.62% and 6.87%) than the leaves of other species. The results from this study indicate that the analyzed species are a rich source of flavonoids, ellagic acid and tannins, which may be used for the quality assessment of Rubus L. species leaves.

The Study of Sugar Analysis in Licorice Extract by HPLC (HPLC를 이용한 감초 추출물의 당 분석법 연구)

  • Kwon Young-Ju;Jang Gi-Chul;Rah Hyo-Hwan;Kim Yong-Ha;Rhee Moon-Su
    • Journal of the Korean Society of Tobacco Science
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    • v.27 no.1 s.53
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    • pp.114-119
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    • 2005
  • The study was carried out to develope quantitative analysis method of individual sugars in licorice extract. Individual sugars were analyzed by HPLC equipped with Refractive Index(RI) Detector. R values of sucrose and glucose were 1.0000 and R values of fructose and maltose were 0.9999. Standard calibration curve showed good linearity. Detection limit of sugars was in the range of 1.58 to 3.92 ${\mu}g$. Recovery rate of fructose, glucose, sucrose and maltose was $99.4\~102.2\%,\;92.3\~97.9\%,\;99.4\~102.0\%,\;91.1\~101.0$ respectively. Measure uncertainty was calculated to confirm trust and accuracy of analytical results. Main uncertainty factors were standard purity and HPLC replication injection. In $95\%$ trust level expanded uncertainty of sugars in licorice extract were fructose $1.98\pm0.047,\;glucose\;1.32\pm0.065,\;sucrose\;11.69\pm1.177,\;maltose\;1.06\pm0.042\;g/100\;g$.

Microanalysis of Metocurine Iodide in Urine and Blood by HPLC (HPLC를 이용한 뇨 및 혈액중 Metocurine Iodide의 미량분석)

  • Kim, Kyung-Nim;Lee, Jong-Pil;Kim, Hyo-Jin;Kim, Bak-Kwang
    • YAKHAK HOEJI
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    • v.38 no.4
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    • pp.379-388
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    • 1994
  • A new method for the analysis of metocurine iodide in biological fluids was developed. Metocurine iodide was quantitatively extracted with rose bengal from aqueous layer into dichloromethane layer and the amount of metocurine iodide was calculated from the amount of rose bengal which was determined by HPLC with fluorescence detector. It was possible to analyze metocurine iodide without the effect of co-prescribed drugs in the concentration range of $0.09{\sim}9.10\;{\mu}g/ml$. The detection limits of metocurine iodide in urine and blood were 0.8 and 1.2 ng at S/N=3, each respectively.

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HPLC Detection of Free Malonaldehyde for Rapid Measurement of Lipid Oxidation Development

  • Key Whang;Kim, Chang-Min
    • Preventive Nutrition and Food Science
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    • v.5 no.1
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    • pp.7-9
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    • 2000
  • Variouss concentrationss of free malonaldehyde were prepared from 1,1,3,3-tetraethoxy propane (TEP). Spectrophoto-metric determination and HPLC analysis of free malonaldehyde instead of malonaldehyde-thiobaribituric acid (MA-TBA) complex were conducted. Malonaldehyde was well separated on a $\mu$Bondapak C18 column. The absorbances at 254 nm and the HPLC peak areas of free malonaldehyde increased with the increase in its concentration. The correlation coefficient between absorbances and peak areas was 0.998. The total time elapsed to conduct the whole procedure was less than 15 minutes. This method directly measured the amount of free malonaldehyde in a short period of time successfully. This procedure is expected to be used as a rapid, accurate and specific means to de-termine the development of lipid oxidation in food.

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Studies on the Chemical Components and Biological Activities of Edible Plants in Korea (II) - Isolation and Quantitative Analysis of Flavonoids from the Leaves of Cedrela sinensis A. Juss. by HPLC - (한국산 식용식물의 화학성분 및 생리활성에 관한 연구 (II) - HPLC에 의한 참죽나무 잎중 Flavonoid 성분의 확인 및 정량 -)

  • 박종철;전순실;양한석;김성환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.5
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    • pp.581-585
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    • 1993
  • Five flavonoids isolated from the ethyl acetate fraction of Cedrela sinensis A. Juss. were identified by high performance liquid chromatography. Separation was achieved by reversed phase chromatography on ${\mu}-bondapak$ C18 column with isocratic elution method. The content of the major flavonoid, quercitrin was about 9.48%(w/w) and 37.06%(w/w) for the methanol extract and ethyl acetate fraction, respectively.

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A Study on Major Components of Bee Venom Using HPLC (HPLC를 이용한 봉약침의 주요 성분에 관한 연구)

  • Lee, Jin-Seon;Kwon, Gi-Rok;Choi, Ho-Young
    • Journal of Acupuncture Research
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    • v.17 no.4
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    • pp.120-129
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    • 2000
  • This study was designed to study on major components of various Bee Venom(Bee Venom by electrical stimulation in Korea; K-BV I, Bee Venom by Microwave stimulation in Korea; K-BV II, 0.5mg/ml, Fu Yu Pharmaceutical Factory, China; C-BV, 1mg/ml, Monmouth Pain Institute, Inc., U.S.A.; A-BV) using HPLC(High performance liquid chromatography). The results were summarized as follows : 1. HPLC method is useful for analysis of Bee Venom when solution rate is above 1:4000. 2. Analysis of Apamin using HPLC, the Retention time was 8.7min, and standard measurement curve was a function of y=4E+06x+21245. 3. Analysis of Melittin using HPLC, the Retention time was 29.0 min, and standard measurement curve was a function of y=4E+06x+23015. 4. Concentration of Melittin was about 297times than Apamin in K-BV I, and about 329times in K-BV II at same 1:500 solution rate, abnormally about 12 times in C-BV at 1:4000 solution rate. 5. Chinese Bee Venom using HPLC, the point from 5 to 7min(Retention time) showed a big extraordinary peak. These data from the study can be applied to establish the standard measurement of Bee Venom and prevent pure bee venom from mixing of another components. I think it is desirable to study more about safety of Bee Venom as time goes by.

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