• Journal of Pharmaceutical Investigation
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• v.21 no.3
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• pp.179-188
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• 1991

An HPLC assay method of a drug to be applied to the pharmacokinetic studies of the drug should be completely validated. The validation process for an HPLC assay method in a biological sample was discussed using the data obtained from the development of HPLC method for the simultaneous quantitation of verapamil and norverapamil in human serum. The validation criteria included were specificity, linearity, accuracy, precision, sensitivity, recovery, drug stability, and ruggedness of an assay method.

• ALGAE
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• v.28 no.3
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• pp.289-296
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• 2013

For the quantitative detection of algal toxin, microcystin, a chemiluminescence immunochromatographic assay method was developed. The developed system consists of four parts, chemiluminescence assay strip (nitrocellulose membrane), horse radish peroxidase labeled microcystin monoclonal antibodies, chemiluminescence substrate (luminol and hydrogen peroxide), and luminometer. The performance of the chemiluminescence immunochromatographic assay system was compared with high performance liquid chromatography (HPLC) detection. The detection limit of chemiluminescence immunochromatographic assay system is several orders of magnitude lower than with HPLC. The chemiluminescence immunochromatography and HPLC results correlated very well with the correlation coefficient ($r^2$) of 0.979.

• Journal of Applied Biological Chemistry
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• v.57 no.4
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• pp.301-305
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• 2014

The Antioxidant activity screening identification of five kind compounds in Artemisiae annuae herba with the on-line screening high performance liquid chromatography (HPLC) $ABTS^+$ assay. The various experimental variables such as the extraction time (h) and extraction solvent composition (%) of dipping method were investigated efficiently extraction at the room temperature $25^{\circ}C$. The results, the highest yield of total extract amount (0.458 g, 15.250%) was obtained by dipping method with 100% water and extraction time to 3 h. And the on-line screening HPLC-$ABTS^+$ assay method was rapid and efficient to search for bioactivity from natural products.

• Analytical Science and Technology
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• v.11 no.6
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• pp.474-477
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• 1998

A simple and rapid method for preparation of standard phosphatidylcholine hydroperoxide (PCOOH) in chemiluminescence-HPLC assay (CL-HPLC) was developed using the rose bengal dependent spectrofluorometric oxidation. The concentration of obtained PCOOH was determined by FOX (ferrous oxidation-xylenol orange) assay and then subjected to a CL-HPLC system.

• KSBB Journal
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• v.29 no.2
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• pp.124-130
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• 2014

Acer tegmentosum was a traditional Korean herbal medicine showing various pharmacological activities. In this work, A. tegmentosum were extracted with boiling water and then successively partitioned with dichloromethane, ethyl acetate, n-butyl alcohol (n-BuOH), and water. Salidoside, the target compound, was purified in n-BuOH phase using a chromatography method. For the analysis of salidoside, TLC and LC-MS were used as well as on-line screening $HPLC-ABTS^+$ assay with three different wavelength of 254, 280, and 320 nm. An amount of 1.34 g of salidoside were obtained in n-BuOH phase fromAcer tegmentosum was a traditional Korean herbal medicine showing various pharmacological activities. In this work, A. tegmentosum were extracted with boiling water and then successively partitioned with dichloromethane, ethyl acetate, n-butyl alcohol (n-BuOH), and water. Salidoside, the target compound, was purified in n-BuOH phase using a chromatography method. For the analysis of salidoside, TLC and LC-MS were used as well as online screening $HPLC-ABTS^+$ assay with three different wavelength of 254, 280, and 320 nm. An amount of 1.34 g of salidoside were obtained in n-BuOH phase from 3 kg of dry biomass. The on-line screening $HPLC-ABTS^+$ assay is rapid and efficient tool to search bioactivity from A. tegmentosum. 3 kg of dry biomass. The on-line screening $HPLC-ABTS^+$ assay is rapid and efficient tool to search bioactivity from A. tegmentosum.

• Natural Product Sciences
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• v.17 no.4
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• pp.315-320
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• 2011

Routinely applicable HPLC assay procedures for the ganglioside content in various deer antler preparations were established through the creation of a UV-absorbing chromophoric substance - trans-${\alpha},{\beta}$-unsaturated-hexadecene-aldehyde - from the sphingosine moiety in ganglioside molecules by two step chemical reactions. In order to guarantee the assay's accuracy and sensitivity, the HPLC-assay procedure adopted internal reference procedures by mixing cis-${\alpha},{\beta}$-unsaturated-hexadecene aldehyde[V] or cis-3-heptadecene- 1,2-diol[IV] to assay samples. The internal reference compound [IV] or [V] was synthesized in our laboratory starting from mannitol-diacetonide through three or four step organic reactions. This new HPLC-assay procedure was successfully applied to deer antler extracts with good dose-dependent calibration curves at the picomole level of gangliosides.

• Analytical Science and Technology
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• v.30 no.3
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• pp.154-158
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• 2017

The Korean Pharmacopoeia (KP XI), British Pharmacopoeia (BP 2013) and Japanese Pharmacopoeia contain monographs for the quality control of raw fusidate sodium and its formulations using high performance liquid chromatography (HPLC). However, the assay method for the determination of fusidate sodium in commercial tablets is titration which is less specific than HPLC. In this study, we present an alternative HPLC method for quantitation of fusidate sodium in tablets. Method validation was performed to determine linearity, precision, accuracy, system suitability, and robustness. The linearity of calibration curves in the desired concentration range was high ($r^2=0.9999$), while the RSDs for intra- and inter-day precision were 0.25-0.37 % and 0.11-0.60 %, respectively. Accuracies ranged from 99.46-100.85 %. Since the system suitability, intermediate-precision and robustness of the assay were satisfactory, this method will be a valuable addition to the Korean Pharmacopoeia (KP XI).

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.381-385
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• 2004

This study was conducted to develop an HPLC method for determining pantothenic acid in fortified foods which has typically been determined by microbiological assay (MBA) according to AOAC and Korean Food Code approved methods. Pantothenic acid was determined by reversed-phase ion-pair HPLC using UV absorption (200 nm) after extraction with 20 mM potassium phosphate solution by sonication. The recovery of spiked samples and detection limit (LOD) by HPLC were 83.5∼109.6% and 0.5 ppm (mg/kg), respectively. The LOD of the microbiological assay (MBA) was much lower than that of HPLC. The concentrations of pantothenic acid analyzed in all tested samples (n=13) confirmed compliance with declared label claims. The range of recovery ratio by the HPLC method when compared to the microbiological assay was 91.9∼117.6%. There was not significant difference (p<0.01) between the HPLC and MBA methods and the equation of the regression curve was y=1.1428x-0.2269 (r=0.9842). This proposed HPLC method for determining pantothenic acid appears to be suitable for determining pantothenic acid concentrations above 0.25 mg/100 g in fortified foods.

• Analytical Science and Technology
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• v.31 no.3
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• pp.107-111
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• 2018

Currently, ultraviolet-visible spectrophotometry and titration methods are used for assay tests of tramadol hydrochloride injection and raw material in the Korean Pharmacopoeia XI (KP XI). Titration has also been used in the British Pharmacopoeia (BP 2013) for the assay test of tramadol hydrochloride, and the HPLC assay for tramadol hydrochloride raw material has been used in the United States Pharmacopeia (USP 39). In this study, we developed an alternative HPLC assay method for tramadol hydrochloride injection that is up to date and specific, and employs the same method as tramadol hydrochloride capsules. Validation of the HPLC method was conducted to determine linearity, precision, accuracy, system suitability, and robustness. The linearity of the calibration curves in the desired concentration range was good ($r^2$ > 0.9999). RSDs of intra-day precision obtained were 0.05-0.08 % and inter-day precision obtained were 0.08-0.19 %. Accuracy was obtained with recoveries in the range of 98.16 % and 100.90 %. As a result of the system's suitability, the RSD of both retention time and the peak area obtained were 0.07 %. The values of the plate number and tailing factor of tramadol hydrochloride obtained were 7076 and 1.16, respectively. Because of the intermediate precision and robustness of the developed assay, it is expected to become a valuable tool for revising the Korean Pharmacopoeia (KP XI).

• Preventive Nutrition and Food Science
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• v.8 no.4
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• pp.301-305
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• 2003

This study was conducted to develop an HPLC method for determining vitamin B$_{12}$ in fortified foods which has typically been determined by microbiological assays according to AOAC and Korean Food Code approved methods. Vitamin B$_{12}$ (cyanocobalamin) was determined by reversed-phase HPLC with a triple column and UV/VIS dectector (550 nm) using the column switching technique after extraction with 5 mM potassium phosphate solution by sonication without a clean-up procedure. The recovery of spiked samples and limit of detection (LOD) by HPLC were 78.6 ∼107.5 % and 2 ppb ($\mu\textrm{g}$/kg), respectively. The LOD of the microbiological assay (MBA) was much lower than that of HPLC. The concentrations of vitamin B$_{12}$ analyzed in all tested samples (n=12) confirmed compliance with declared label claims. The range of recovery ratio by the HPLC method when compared to the microbiological assay was 76.2 ∼140.0 %. There was not significant difference between the HPLC and MBA methods (p < 0.01) with r=0.9791 and linear regression y=0.9923x-0.04. The HPLC method for determining vitamin B$_{12}$ using the column-switching technique appears to be suitable for determining vitamin B$_{12}$ concentrations above 1 $\mu\textrm{g}$/100 g in fortified foods.ied foods.