• Natural Product Sciences
• /
• v.24 no.1
• /
• pp.36-39
• /
• 2018

Phytochemical analysis of Boehmeria nivea (Bn) leaves by medium pressure liquid chromatography led to the isolation of a flavonoid glycoside identified by spectroscopic analysis as rutin. The amount of rutin in the leaves of Bn harvested from nine regions in South Korea (Bn 1-9) which were collected on the months of June, July, August, and September was determined by HPLC-UV analysis. A gradient elution program that utilizes a $Discovery^{(R)}$ C18 ($4.6{\times}250mm$, $5{\mu}m$) column and mobile phase composed of 1% acetic acid-water: acetonitrile (90:10 to 60:40 for min) was followed. The injection volume and flow rate were $10{\mu}l$ and 1 mL/ min, respectively. UV detection was set at 350 nm. Results show that Bn-8 harvested in September reported the highest content of rutin among the samples analyzed. This study provides a basis for the optimal harvest time of Bn which maximizes the yield of rutin.

• Korean Journal of Pharmacognosy
• /
• v.49 no.1
• /
• pp.70-75
• /
• 2018

Acer tegmentosum Maxim (ATM) has been used to treat hepatic disorders in traditional oriental medicine. However, there is little information about phytochemical constituents for quality control of ATM. In this study, we developed and established a simultaneous analytical method of the 10 marker compounds (three coumarins, 3 flavonoids, 1 lignan, 3 phenolics) in ATM using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). Chromatographic separation of ten target analytes was achieved with a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$), using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. Identifications and quantitation of all analytes were performed using a Q-Exactive UPLC-MS/MS system. Correlation coefficients of the calibration curve for all analytes were ${\geq}0.9986$. The values of limits of detection and quantification of all analytes were 0.5-10.0 and 5.0-50.0 ng/mL, respectively. The established UPLC-MS/MS method successfully identified all target analytes in ATM, and the phytochemicals were 0.01-67.98 mg/g in its lyophilized water extract.

• Mass Spectrometry Letters
• /
• v.11 no.1
• /
• pp.10-14
• /
• 2020

Riboflavin is a water-soluble vitamin, which serves as a precursor to flavin mononucleotide and flavin adenine dinucleotide. This study aimed to develop a simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for the quantification of riboflavin in the Beagle dog plasma. This method utilized simple protein precipitation with acetonitrile and ¹³C₄, ¹⁵N₂-riboflavin was used as an internal standard (IS). For chromatographic separation, a hydrophilic interaction liquid chromatography (HILIC) column was used with gradient elution. The mobile phase consisted of 0.1% (v/v) aqueous formic acid with 10 mM ammonium formate and acetonitrile with 0.1% (v/v) formic acid. Since riboflavin is an endogenous compound, 4% bovine serum albumin in phosphate buffered saline was used as a surrogate matrix to prepare the calibration curve. The quantification limit for riboflavin in the Beagle dog plasma was 5 ng/mL. The method was fully validated for its specificity, sensitivity, accuracy and precision, recovery, and stability according to the US FDA guidance. The developed LC-MS/MS method may be useful for the in vivo pharmacokinetic studies of riboflavin.

• Natural Product Sciences
• /
• v.23 no.4
• /
• pp.270-273
• /
• 2017

A quantitative analysis of bakkenolide D in the different parts of Petasites japonicus and Farfugium japonicum was performed by HPLC. A gradient HPLC elution system with a mobile phase consisting of water: acetonitrile solution (20:80 to 0:100 for 45 min) was followed and an INNO $C_{18}$ column was used for the chromatographic separation. The injection volume, flow rate, and UV detection were $10{\mu}L$, 1 mL/min, and 290 nm, respectively. Results show that both species showed the highest amount of bakkenolide D in the roots being 107.203 and 166.103 mg/g for P. japonicas and F. japonicum, respectively. Content analysis on the different parts of both plants displayed remarkably lower values which ranged from 0.403 - 4.419 and 7.252 - 32.614 mg/g for P. japonicas and F. japonicum, respectively. The results show that the roots of both plants are rich in bakkenolide D showing a promising use in the development of nutraceuticals and industrial application of the compound.

• Korean Journal of Pharmacognosy
• /
• v.46 no.4
• /
• pp.352-364
• /
• 2015

The aim of this study was to quantitatively analyze for quality assessment of eighteen marker compounds, including homogentisic acid, 3,4-dihydroxybenzaldehyde, spinosin, liquiritin, hesperidin, ginsenoside Rg1, liquiritigenin, ginsenoside Rb1, glycyrrhizin, 6-gingerol, atractylenolide III, honokiol, costunolide, dehydrocostuslactone, atractylenolide II, nootkatone, magnolol, and atractylenolide I, in Hyangsayukgunja-tang using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer. The column for separation of eighteen marker components were used a UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}$) and kept at $45^{\circ}C$ by gradient elution with 0.1% (v/v) formic acid in water and acetonitrile as mobile phase. The flow rate and injection volume were 0.3 mL/min and $2.0{\mu}l$, respectively. The correlation coefficient of all marker compounds was ${\geq}0.9914$, which means good linearity, within the test ranges. The limits of detection and quantification values of the all analytes were in the ranges 0.04-1.11 and 0.13-3.33 ng/mL, respectively. As a result, five compounds, homogentisic acid, 3,4-dihydroxybenzaldehyde, spinosin, liquiritigenin, and atractylenolide I, in this sample were not detected and the amounts of the 13 compounds except for the 5 compounds were $8.10-6736.37{\mu}g/g$ in Hyangsayukgunja-tang extract.

• Korean Journal of Pharmacognosy
• /
• v.46 no.4
• /
• pp.342-351
• /
• 2015

In this study, we performed quantitative determination of the three marker compounds such as gallic acid, ellagic acid, and quercetin in the 70% ethanol extracts of non-processed Sanguisorbae Radix and processed Sanguisorbae Radix using a high-performance liquid chromatography coupled with photodiode array detector. The analytical column for separation of the three compounds was used a Gemini $C_{18}$ column ($5{\mu}m$, $4.6{\times}250mm$) by the gradient elution with distilled water and acetonitrile containing 1.0% (v/v) acetic acid as mobile phase. The flow rate and injection volume were $1.0{\mu}L/min$ and $10{\mu}L$. The concentrations of gallic acid, ellagic acid, and quercetin in non-processed Sanguisorbae Radix were 0.25, 0.26, and 0.007%, respectively, while the concentrations of gallic acid, ellagic acid, and quercetin in non-processed Sanguisorbae Radix 0.14-0.55, 0.27-2.03, and 0.001-0.007%, respectively. Among the three components, the amount of the ellagic acid was increased after processing in Sanguisorbae Radix.

• Analytical Science and Technology
• /
• v.16 no.6
• /
• pp.499-503
• /
• 2003

A high performance liquid chromatography gradient elution method with fluorescence detection for the determination of biotin in pure form and pharmaceutical preparations has been developed. BrMDMC gives intense fluorescence and the fluorescence was monitored with excitation at 360 nm and emission at 410 nm. The calibration curve for biotin shows good linearity over the range of 5 ~ 400 ng with correlation coefficient of 0.999. The detection limit of biotin was 2 ng and the result of recovery was 98.75% with relative standard deviation of 1.1%.

• Analytical Science and Technology
• /
• v.11 no.6
• /
• pp.429-435
• /
• 1998

Free and glycine conjugated bile acids were detected fluoromatrically in high-performance liquid chromatography after derivatization with dansyl cadaverine. The coupling agent, diethyl phosphorocyanidate was used to form the amide bond between dansyl cadaverine and analytes. The dansyl derivatives of 8 bile acids were separated successfully on Cosmosil ODS column by using linear gradient elution of 20% MeOH-water/$CH_3CN$. The detection limits of cholic acid was reached 10 pg(S/N=5) per $1{\mu}l$ of injected volume. This new derivatization method would be applicable to detect the changes of bile acids in biological samples.

• Food Science and Biotechnology
• /
• v.18 no.2
• /
• pp.561-564
• /
• 2009

A high performance liquid chromatography (HPLC) method has been developed for determination of 11 ginsenosides in black ginseng (BG, white ginseng that is subjected to 9 cycles of $95^{\circ}C$ for 3 hr). After eluted by gradient elution of water-acetonitrile without buffer in 70 min, 11 ginsenosides in BG were identified. The proposed method provided good linearity ($R^2$>0.9995), accuracy (92.2-106.6%), and intra- and interday precision (RSD<2.6%). In addition, ginsenosides compositions in white, red, and black ginsengs were investigated using this method, respectively. Interestingly, in BG, the content of ginsenoside $Rg_3$ which does not existed in white ginseng was 7.51 mg/g, approximately 20 times than that in red ginseng.

• Bulletin of the Korean Chemical Society
• /
• v.35 no.1
• /
• pp.197-203
• /
• 2014

A rapid, accurate and precise method for the determination of 22 amino acids in Isatidis Radix by Hydrophilic Interaction Ultra-High-Performance Liquid Chromatography Coupled with Triple-Quadrupole Mass Spectrometry (HILIC-UPLC-MS/MS) was established. Chromatographic separation was carried out on a Acquity UPLC BEH Amide column ($2.1mm{\times}100mm$, $1.7{\mu}m$) with gradient elution of acetonitrile (containing 0.05% formic acid and 2 mM ammonium formate) and water (containing 0.15% formic acid and 10 mM ammonium formate) at a flow rate of 0.4 mL/min; Waters Xevo$^{TM}$ TQ worked in multiple reaction monitoring mode. All components were separated in 17 min. All calibration curves were linear ($R^2$ > 0.991) over the tested ranges. The limits of detection (LOD) and limits of quantitation (LOQ) for these compounds were 0.21-79.55 and 0.72-294.23 ng/mL, respectively. The average recoveries were in the range of 93.75-104.16% with RSD value less than 6.56%. Therefore, this method could be an alternative assay for the determination of 22 amino acids in Isatidis Radix due to its rapidness, sensitivity, less sample and solvent consumption.