• Title/Summary/Keyword: method validation #5

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Quantitative Analysis of Glycine Semen Nigra and Eucommiae Cortex for Standardization of GCSB-5 Preparation (생약복합제 GCSB-5의 품질 표준화를 위한 흑두 및 두충의 함량 분석)

  • Lee, Eun-Hee;Cha, Bae-Cheon
    • Korean Journal of Pharmacognosy
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    • v.40 no.1
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    • pp.18-24
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    • 2009
  • GCSB-5 preparation is a purified extract from a mixture six herbal medicines (Acanthopanacis Cortex, Achyranthis Radix, Saposhnikoviae Radix, Cibotii Rhizoma, Glycine Semen Nigra, Eucommiae Cortex) that have been widely used in traditional medicine to treat various bone disorders. This study was carried out to obtain the HPLC analysis method that can be used to establish quantitative analysis of Glycine Semen Nigra and Eucommiae Cortex for standardization of GCSB-5 preparation. HPLC analysis methods for the simultaneous determination of genistin (Glycine Semen Nigra) and geniposide (Eucommiae Cortex) were established for the quality control of herbal medicinal raw material and preparation. And validation of HPLC analysis methods were conformed for verification of HPLC methods by check to specificity, linearity, intra-day precision, inter-day precision and accuracy following ICH guideline. As the result of quantitative analysis, the contents of genistin and geniposide in the raw material of GCSB-5 preparation were 0.0426-0.0427 mg/g and 0.431-0.432 mg/g. And GCSB-5 preparation contained genistin of 0.0202-0.0203 mg/capsule and geniposide of 0.211-0.212 mg/capsule, respectively.

Non-destructive Method for Selection of Soybean Lines Contained High Protein and Oil by Near Infrared Reflectance Spectroscopy

  • Choung, Myoung-Gun;Baek, In-Youl;Kang, Sung-Taeg;Han, Won-Young;Shin, Doo-Chull;Moon, Huhn-Pal;Kang, Kwang-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.5
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    • pp.401-406
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    • 2001
  • The applicability of non-destructive near infrared reflectance spectroscopic (NIRS) method was tested to determine the protein and oil contents of intact soybean [Glycine max (L.) Merr.] seeds. A total of 198 soybean calibration samples and 101 validation samples were used for NIRS equation development and validation, respectively. In the developed non-destructive NIRS equation for analysis of protein and oil contents, the most accurate equation was obtained at 2, 8, 6, 1(2nd derivative, 8 nm gap, 6 points smoothing, and 1 point second smoothing) and 2, 1, 20, 10 math treatment conditions with Standard Normal Variate and Detrend (SNVD) scatter correction method and entire spectrum (400-2500 nm) by using Modified Partial Least Squares (MPLS) regression, respectively. Validation of these non-destructive NIRS equations showed very low bias (protein: 0.060%, oil: -0.017%) and standard error of prediction (SEP, protein: 0.568 %, oil : 0.451 %) as well as high coefficient of determination ($R^2$, protein: 0.927, oil: 0.906). Therefore, these non-destructive NIRS equations can be applicable and reliable for determination of protein and oil content of intact soybean seeds, and non-destructive NIRS method could be used as a mass screening technique for selection of high protein and oil soybean in breeding programs.

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Simultaneous Analysis of three Marker Components in Hwangryunhaedok-tang by HPLC-DAD (황련해독탕 중 3종 생리활성 물질의 HPLC-DAD 동시 정량분석법 확립)

  • Yang, Hye-Jin;Weon, Jin-Bae;Ma, Jin-Yeul;Ma, Choong-Je
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.64-68
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    • 2011
  • In this study, a high performance liquid chromatography-diode array detector method was established, for simultaneous determination of three compounds, berberine, palmatine and geniposide in Hwangryunhaedok-tang, To develop and validate method, $C_{18}$ column (5 ${\mu}M$, 4.6 mm${\times}$250 mm) was used with gradient mobile phase, water containing 0.1% trifluoroacetic acid (TFA) and MeOH at the column temperature of $30^{\circ}C$. UV wavelength was set at 230 and 280 nm. Validation of the chromatography method was evaluated by linearity, precision and accuracy test. Calibration curve of standard components showed good linearity ($R^2$ > 0.9999). The limits of detection (LOD) and limits of quantification (LOQ) varied from 0.05 to 0.17 ${\mu}g/ml$ and 0.15 to 0.53 ${\mu}g/ml$, respectively. The relative standard deviations (RSDs) data of intra-day and inter-day test were in less than 2.99% and 1.90%, respectively. The results of the accuracy test were in the range of 98.36 to 102.52% with RSDs values 0.32 to 1.98%. The results of validation indicated that this method was a very accurate and sensitive assay.

Simultaneous Determination of Platyphylloside, Aceroside VIII and Betulin in Betula platyphylla bark by HPLC-DAD (HPLC-DAD를 이용한 자작나무 수피의 Platyphylloside, Aceroside VIII 및 Betulin의 동시분석법 확립)

  • Cho, NamKi;Kim, Dae Hyun;Sung, Sang Hyun
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.294-299
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    • 2014
  • The simultaneous determination of platyphylloside, aceroside and betulin was established for the quality control of Betula platyphylla bark using a high performance liquid chromatography and diode-array UV/Vis detector (HPLC-DAD). Separation and quantification were successfully achieved with a INNO C18 column ($5{\mu}m$, 4.6 mm $I.D.{\times}150mm$) by gradient elution of a mixture of methanol and water at a flow rate of 1.0 ml/min. Validation of the developed method was performed by various factor such as linearity, specificity, precision, accuracy, system suitability and stability. This method was successfully applied to the determination of contents of platyphylloside, aceroside VIII and betulin in three batches of Betula platyphylla bark extract. These results suggest that the developed HPLC method is simple, effective and could be utilized as a quality control method for Betula platyphylla bark products.

Validation of a Simple HPLC Method for Determination of Ciprofloxacin Hydrochloride in Human Plasma (인체 혈장에서 염산시프로플록사신(시프로플록사신으로서 250 mg) 정량을 위한 HPLC 분석법의 유효성검토)

  • Ha, Yong-Hwa;Cho, Sung-Hee;Chun, Sung-Kuk;Seo, Seong-Hoon;Rew, Jae-Hwan;Choi, Young-Wook;Lee, Kyung-Tae
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.327-331
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    • 2004
  • A simple HPLC method with ultraviolet detection of ciprofloxacin in human plasma was developed and validated. After protein precipitation with trichloroacetic acid, chromatographic separation of ciprofloxacin in plasma was achieved at $50^{\circ}C$ with a $C_{18}$ column and methanol-phosphate mixture (pH 2.5), as mobile phase. Quantitative determination was performed by ultraviolet detection at 278 nm. The method was specific and validated with a limit of quantification of 100 ng/ml. The intra- and inter-day coefficients of variation were between 1.67% and 10.55% and accuracy between 92.01 % and 106.09%. The method has been successfully applied in a bioavailability study of 250 mg ciprofloxacin hydrochloride tablet.

Analytical Method Validation of (-)Epicatechin gallate in Penthorum chinense Pursh Extract using HPLC

  • Kwon, Jin Gwan;Jung, Yeon Woo;Seo, Changon;Hong, Seong Su;Lee, Ji Eun;Shin, Hyun Tak;Jung, Su Young;Choi, Chun Whan;Kim, Jin Kyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.100-100
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    • 2019
  • This study attempted to establish a High Performance Liquid Chromatography (HPLC) analysis method for the determination of (-)-epicatechin gallate as a part of the quality control for the development of functional cosmetic materials from Penthorum chinense Pursh extracts. HPLC was performed on a Unison US-C18 column ($4.6{\times}250mm$, $5{\mu}m$) with a gradient elution of 0.05% (v/v) trifluoroacetic acid (TFA) and methyl alcohol at a flow rate of 1.0 mL/min at $30^{\circ}C$. The analyte was detected at 280 nm. The HPLC method was performed in accordance with the International Conference on Harmonization (ICH) guideline (version 4, 2005) of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation were 0.11 and 0.33 mg/mL, respectively. Calibration curves showed good linearity (r2 > 0.9999), and the precision of analysis was satisfied (less than 0.6%). Recoveries of quantified compounds ranged from 99.51 to 101.92%. This result indicates that the established HPLC method is very useful for the determination of marker compound in P. chinense Pursh extracts.

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Validation of an HPLC/UV-based method for Salicornia herbacea-derived isorhamnetin-3-O-glucoside and quercetin-3-O-glucoside quantification

  • Park, Jun Yeon;Paje, Leo Adrianne;Kang, Ki Sung;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.64 no.3
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    • pp.285-290
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    • 2021
  • Salicornia herbacea is a type of salt marsh plant that has been used in traditional medicine to treat several diseases. Isorhamnetin-3-O-glucoside (I3G) and quercetin-3-O-glucoside (Q3G) are major flavonoids in S. herbacea that are known to exert various pharmacological activities. Therefore, our study sought to validate and optimize an HPLC/UV-based analytical method for I3G and Q3G yield quantification, as well as to determine its limit of detection, limit of quantification, linearity, precision, and accuracy. Upon testing a concentration range of 31.5-1.9 ㎍/mL the results exhibited good linearity (r2 ≥0.9996 and r2 ≥0.9999 for I3G and Q3G, respectively), and the procedure was deemed precise (relative standard deviation of ≤3.19 and ≤3.85%, respectively), and accurate (102.6-105.0 and 92.9-95.2%, respectively). The results showed that our proposed method could be used for rapid I3G and Q3G evaluation in S. herbacea.

Estimation of the Hapcheon Dam Inflow Using HSPF Model (HSPF 모형을 이용한 합천댐 유입량 추정)

  • Cho, Hyun Kyung;Kim, Sang Min
    • Journal of The Korean Society of Agricultural Engineers
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    • v.61 no.5
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    • pp.69-77
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    • 2019
  • The objective of this study was to calibrate and validate the HSPF (Hydrological Simulation Program-Fortran) model for estimating the runoff of the Hapcheon dam watershed. Spatial data, such as watershed, stream, land use, and a digital elevation map, were used as input data for the HSPF model. Observed runoff data from 2000 to 2016 in study watershed were used for calibration and validation. Hydrologic parameters for runoff calibration were selected based on the user's manual and references, and trial and error method was used for parameter calibration. The $R^2$, RMSE (root-mean-square error), RMAE (relative mean absolute error), and NSE (Nash-Sutcliffe efficiency coefficient) were used to evaluate the model's performance. Calibration and validation results showed that annual mean runoff was within ${\pm}4%$ error. The model performance criteria for calibration and validation showed that $R^2$ was in the rang of 0.78 to 0.83, RMSE was 2.55 to 2.76 mm/day, RMAE was 0.46 to 0.48 mm/day, and NSE was 0.81 to 0.82 for daily runoff. The amount of inflow to Hapcheon Dam was calculated from the calibrated HSPF model and the result was compared with observed inflow, which was -0.9% error. As a result of analyzing the relation between inflow and storage capacity, it was found that as the inflow increases, the storage increases, and when the inflow decreases, the storage also decreases. As a result of correlation between inflow and storage, $R^2$ of the measured inflow and storage was 0.67, and the simulated inflow and storage was 0.61.

LC-MS/MS Method for Simultaneous Analysis of Growth Hormone-Releasing Peptides and Secretagogues in Human Urine

  • Min, Hophil;Han, Boyoung;Sung, Changmin;Park, Ju-Hyung;Lee, Kang Mi;Kim, Ho Jun;Kim, Ki Hun;Son, Junghyun;Kwon, Oh-Seung;Lee, Jaeick
    • Mass Spectrometry Letters
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    • v.7 no.3
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    • pp.55-63
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    • 2016
  • Growth hormone (GH)-releasing peptides (GHRPs) and GH secretagogues (GHSs) are listed in the World Anti-Doping Agency (WADA) Prohibited List. In the present study, we developed and validated a method for the simultaneous analysis of seven GHRPs (alexamorelin, GHRP-1, -2, -4, -5, -6, and hexarelin) and three GHSs (anamorelin, ibutamoren, and ipamorelin) in human urine. Method validation was performed at minimum required performance levels specified by WADA technical documents (2 ng/mL) for all substances, and the method was validated with regard to selectivity (no interference), linearity (R2 > 0.9986), matrix effects (50.0%-141.2%), recovery (10.4%-100.8%), and intra- (2.8%-16.5%) and inter-day (7.0%-22.6%) precisions. The limits of detection for screening and confirmation were 0.05-0.5 ng/mL and 0.05-1 ng/mL, respectively.

Validation of Non-invasive Method for Electrocardiogram Recording in Mouse using Lead II

  • Kim, Myung Jun;Lim, Ji Eun;Oh, Bermseok
    • Biomedical Science Letters
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    • v.21 no.3
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    • pp.135-143
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    • 2015
  • Electrocardiogram measures the electric impulses generated by the heart during its cycle. Recently genome-wide association studies on electrocardiogram traits revealed many relevant genetic loci. Therefore, these findings need to be validated and investigated to determine the underlying mechanisms using mouse models. Invasive radiotelemetry has been widely used to record the electrocardiogram in mice because it has several advantages over non-invasive measurements. However, radiotelemetry is expensive and requires complicated surgery. On the other hand, a non-invasive method using 3 electrodes (one for earth) for lead II is easy to establish and allows for rapid measurement. In this study, eleven mice were measured with this non-invasive method and no statistical difference among them was found in any ECG measurements. In addition, repeat measurement in the same mouse was performed in 9 sets of experiment and the results indicated that non-invasive method was reliable for reproducibility. Further it was shown that measurements for 1, 5, 10, and 15 minutes were not different so that a short recording such as 5 minutes was enough to estimate the ECG values including heart rate. Further this method was validated by measuring the ECG of Balb/c and FVB that were previously shown to differ in ECG values by radiotelemetry. Significant differences were found in heart rate, PR interval and corrected QT interval between these mouse strains. This study partially proved that non-invasive method also could provide the accuracy and reproducibility. Based on these results, the non-invasive ECG recordings of lead II is recommended as a useful method for quick test in mouse model.