• Korean Journal of Clinical Pharmacy
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• v.15 no.1
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• pp.46-49
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• 2005

A simple HPLC method was employed for the determination of pentoxifylline in human serum. After addition of internal standard (IS, 50 uL of 3 ug/mL chloramphenicol methanol solution) into the serum sample, the drug and IS were extracted by dichloromethane. Following a 1-min vortex-mixing and a 15-min centrifugation at 3500 게m, the organic phase was transferred and evaporated to dryness under a vacuum. The residue was reconstituted with 120 ${\mu}L$ of mobile phase and 50 ${\mu}L$ was injected into C18 column with a mobile phase composed of 0.034 M phosphoric acid adjusted to pH 4 with 10 M NaOH and acetonitrile (75:25, v/v). The samples were detected using an ultraviolet detector at 273 nm. The method was simple, specific and validated with a limit of 10 ng/mL. Intra- and inter-day precision and accuracy were acceptable for all quality control samples including the lower limit of quantification. The applicability of this method was evaluated by analysis of human serum after oral administration of a single 400 mg dose to 8 healthy subjects. The pharmacokinetic parameters for pentoxifylline in human subjects were calculated using WinNonlin program. As a result, $AUC_{t},\;C_{max},\;T_{max}$ and $t_{1/2}$ were $962.28{\pm}645.69\;ng{\cdot}/mL$, $132.82{\pm}42.05$ ng/mL, $2.06{\pm}2.68$ hr and $8.74{\pm}4.38$ hr, respectively. Based on the results, this validated method appears to be useful fur the pharmnacokinetic study of pentoxifylline in humans.

• Biomolecules & Therapeutics
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• v.13 no.2
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• pp.118-122
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• 2005

The objective of the present investigation was to study pharmacokinetics of promethazine in Korean healthy subjects using a validated HPLC method. The HPLC analysis was performed on a Capcell Pak CN column with a mixture of acetonitrile-0.02M potassium dihydrogen phosphate (42:58, v/v, pH 6.0) and the analyte was quantified with UV detection at 251 nm. The calibration curve of the drug was linear over the range of 1-40ng/mL in human serum and the limit of quantification (LOQ) was 1 ng/mL. This analytical method was validated and shown to be specific, accurate, precise and reproducible. This method was applied to pharmacokinetic study of promethazine in Korean healthy volunteers following an oral administration of two 25 mg Himazin tablets (50 mg promethazine ${\cdot}$HCI) after overnight fasting. Serum samples were collected at given intervals over a 36-hour period (12 points) and pharmacokinetic parameters were determined from serum concentration-time profile using WinNonlin program. The estimated $AUC_{0__\infty}$, $AUC_{0_\infty}$, $C_{max}$, $T_{max}$ and $t_{1/2}$ of promethazine obtained from Korean healthy subjects were 103.84 ${\pm}$84.30 ng${\cdot}$hr/mL, 87.94${\pm}$81.02 ng${\cdot}$hr/mL, 13.43${\pm}$10.92 ng/mL, 2.00${\pm}$1.16 hr and 5.88${\pm}$3.47 hr, respectively.

• The Korea Journal of Herbology
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• v.25 no.3
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• pp.65-71
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• 2010

Objectives：To develop and validate HPLC-PDA methods for simultaneous determination of seven constituents in Samchulkunbi-tang (SKT). Additionally, we investigated the cytotoxicity against BEAS-2B cell line and splenocytes of SKT. Methods：Reverse-phase chromatography using a Gemini $C_{18}$ column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 230, 254 and 280 nm, were used for quantification of the three marker components of SKT. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. The cytotoxicity of SKT were measured by the CCK-8 assay method. Results：Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD) values (%) for intra- and inter-day precision were less than 6.0%. The recovery rate of each compound was in the range of 86.89-109.78%, with an RSD less than 4.0%. The contents of seven compounds in SKT were 1.39-6.84 mg/g. SKT had no cytotoxicity effect at 50-200 ${\mu}g$/mL concentrations. Conclusions：The established method will be helpful to improve quality control and in vitro efficacy study of SKT.

• Korean Journal of Pharmacognosy
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• v.38 no.2 s.149
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• pp.192-196
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• 2007

A rapid and practical HPLC assay was developed for quantitative analysis of saponins in Platycodi Radix. Seven saponin components in Platycodi Radix, i.e., deapioplatycoside E, platycoside E, deapioplatycodin D$_3$, platycodin D$_3$, polygalacin D$_2$, platycodin D$_2$ and platycodin D were successfully resolved on C18 column and detected by ELSD. Standard curves were linear over the concentration range 1 ${\sim}$2,000 ${\mu}$g/ml (r$^2$>0.992). Intra- and inter-day coefficients for variation of seven saponin components were < 10% and limit of quantification of them were around 0.7${\sim}$1.5 ${\mu}$g/ml, respectively. Using this method, contents of seven saponins in various plant materials under different cultivating conditions were estimated.

• Korean Journal of Pharmacognosy
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• v.48 no.3
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• pp.232-236
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• 2017

Gastrodia elata has been widely used as a traditional medicinal herb. However, in Korean Pharmacopeia, neither a marker constituent nor an analytical procedure for G. elata has been established. In this study, we suggest gastrodin and gastrodigenin as marker constituents of G. elata, and propose an analytical procedure for simultaneous quantification of these constituents by high-performance liquid chromatography-ultraviolet spectroscopy (HPLC-UV). The analytical method was validated for its linearity, precision, accuracy, and specificity. Based on the validated method, gastrodin and gastrodigenin in 14 commercial G. elata samples were quantified.

• Toxicological Research
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• v.31 no.3
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• pp.299-305
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• 2015

An HPLC analytical method was validated for the quantitative determination of biogenic amines in agricultural products. Four agricultural foods, including apple juice, Juk, corn oil and peanut butter, were selected as food matrices based on their water and fat contents (i.e., non-fatty liquid, non-fatty solid, fatty liquid and fatty solid, respectively). The precision, accuracy, recovery, limit of detection (LOD) and quantification (LOQ) were determined to test the validity of an HPLC procedure for the determination of biogenic amines, including tryptamine, ${\beta}$-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, in each matrix. The LODs and LOQs for the biogenic amines were within the range of 0.01~0.10 mg/kg and 0.02~0.31 mg/kg, respectively. The relative standard deviation (RSD) of intraday for biogenic amine concentrations ranged from 1.86 to 5.95%, whereas the RSD of interday ranged from 2.08 to 5.96%. Of the matrices spiked with biogenic amines, corn oil with tyramine and Juk with putrescine exhibited the least accuracy of 84.85% and recovery rate of 89.63%, respectively, at the lowest concentration (10 mg/kg). Therefore, the validation results fulfilled AOAC criteria and recommendations. Subsequently, the method was applied to the analysis of biogenic amines in fermented agricultural products for a total dietary survey in Korea. Although the results revealed that Korean traditional soy sauce and Doenjang contained relatively high levels of histamine, the amounts are of no concern if these fermented agricultural products serve as condiments.

• Analytical Science and Technology
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• v.31 no.2
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• pp.88-95
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• 2018

A simple, sensitive, and selective high-performance liquid chromatography (HPLC) method was developed and validated for the simultaneous quantification of fluorometholone (FLU) and tetrahydrozoline hydrochloride (THZ) in combined ophthalmic formulations. The effects of several factors, including detection wavelength, buffer pH, buffer concentration, and organic solvent concentration, on the performance of the method were investigated. The performance of the developed method was validated in accordance with the requirements of the International Conference on Harmonization. The linearity of the calibration curves in the desired concentration range was high for both FLU and THZ ($r^2=0.9999$). The accuracy and precision of the method were determined by a recovery study at analyte concentrations of 80 %, 100 %, and 120 %. The recovery was 99.60-100.17 % for both analytes, with a relative standard deviation less than 1.0 % at any concentration level. The validated results indicate that the new HPLC method could be successfully applied for the simultaneous determination of FLU and THZ in ophthalmic suspensions.

• Herbal Formula Science
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• v.21 no.1
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• pp.71-79
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• 2013

Objectives : Yijin-tang has been used in the treatment of gastrointestinal diseases such as irritable bowel syndrome, gastritis, and gastric ulcer. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous analysis of six compounds, liquiritin, glycyrrhizin, hesperidin, 6-gingerol, homogentisic acid, and 3,4-dihydroxybenzaldehyde in Yijin-tang, a traditional Korean herbal medicinal preparation. Methods : A Gemini C18 column was used for the separation of six constituents at $40^{\circ}C$. The mobile phase using gradient elution consist of two solvent systems, 1.0% acetic acid in water (A) and 1.0% acetic acid in acetonitrile (B). The flow-rate was 1.0 mL/min and injection volume was $10{\mu}g$. The detector was a photodiode array (PDA) detector set at 254 nm and 280 nm. Results : The calibration curves of six compounds showed good linearity in various concentration ranges ($R^2{\geq}0.9997$). The limits of detection (LOD) and limits of quantification (LOQ) were 0.028-$0.192{\mu}g/mL$ and 0.093-$0.540{\mu}g/mL$, respectively. The RSD (%) of the intra and inter day validations were 0.03-0.84% and 0.05 -1.00%, respectively. Recovery was 96.14-01.90% and RSD (%) was less than 1.5%. Conclusions : The established simultaneous analysis methods will help management to improve the quality of Yijin-tang.

• Natural Product Sciences
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• v.18 no.1
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• pp.39-46
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• 2012

In Korea, the leaves of Sonchus brachyotus (Compositae), an edible mountainous vegetable, are traditionally used to treat hepatitis and hemorrhage and are known to have diuretic action. The aqueous ethanolic extract of this plant was selected in our screening experiment using the peroxynitrite ($ONO_2^-$)-scavenging assay, and the present study was performed to qualitatively and quantitatively identify the active compounds from S. brachyotus and validate the present high-permormance liquid chromatography (HPLC) coupled with ultraviolet absorption detection method based on accuracy, precision and repeatability. Five phenolic substances including the main compound, luteolin $7-O-{\beta}-D$-glucuronopyranoside, as well as chlorogenic acid, luteolin 7-O-rutinoside, luteolin $7-O-{\beta}-D$-glucopyranoside, and luteolin, were found in the aqueous ethanolic extract of S. brachyotus. In the HPLC validation experiment, the linearity of the four compounds was established by $R^2$ values of more than 0.999 within the test ranges, and the recovery rate ranged from 98.2 - 105.3%. Luteolin 7-O-glucuronide was a predominant compound (143 mg/g of extract and 18.3 mg/g of the dry weight of plant material) with a potent peroxynitrite-scavenging effect ($IC_{50}$, $1.02{\pm}0.08{\mu}M$). Luteolin and its three glycosides together with chlorogenic acid were qualitatively and quantitatively determined using an HPLC method validated in the present study.

• Natural Product Sciences
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• v.19 no.1
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• pp.20-27
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• 2013

A simple high performance liquid chromatography - diode array detector (HPLC-DAD) method has been developed and validated for simultaneous determination of the six components (cinnamic acid, cinnamaldehyde, eugenol, atractylenolide I, atractylenolide III, and ergosterol) in Oryeongsan. In addition, identification of six marker compounds was conducted by a LC-MS/MS analysis. The six compounds in Oryeongsan were separated on Shishedo $C_{18}$ column (5 ${\mu}m$, $4.6{\times}250$ mm) at a column temperature of $30^{\circ}C$. The mobile phase was a mixture of 0.1% trifluoroacetic acid (TFA) water and acetonitrile employing gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was set at 205, 250, 280, and 330 nm. The developed method had good linearity ($R^2$ > 0.9997) and the limit of detection (LOD) and limit of quantification (LOQ) were observed within the ranges 0.01~0.15 and 0.05~0.45 ${\mu}g/mL$, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were indicated that less than 3% and 90.31~103.31% of accuracy. The results of recovery test were 90.56~106.72% and RSD range was measured from 0.84 to 2.95%. In conclusion, this HPLC-DAD method has been successfully applied to the simultaneous determination of six compounds in Oryeongsan samples.