Simultaneous Determination of Four Compounds, Campesterol, Emodin8-O-β-D-Glucopyranoside, Quercetin, and Isoquercitrin in <i>Reynoutria sachalinensis</i> by High-performance Liquid Chromatography-Diode Array Detector

Eom, Min Rye,Weon, Jin Bae,Jung, Youn Sik,Ryu, Ga Hee,Yang, Woo Seung,Ma, Choong Je Medknow PublicationsMedia Pvt Ltd 2017 Pharmacognosy magazine Vol.13 No.50

<P><B>Background:</B></P><P><I>Reynoutria sachalinensis</I> is a well-known and used herbal medicine to treatment of arthralgia, jaundice, amenorrhea, coughs, carbuncles, and sores.</P><P><B>Objective:</B></P><P>We have developed high-performance liquid chromatography analysis method for simultaneous determination of isolated four compounds, campesterol, emodin8-O-β-D-glucopyranoside, quercetin, and isoquercitrin from <I>R. sachalinensis</I> is.</P><P><B>Materials and Methods:</B></P><P>The four compounds were separated on Shiseido C<SUB>18</SUB> column (S-5 μm, 4.6 mm I.D. ×250 mm) at a column temperature of 25°C. The mobile phase composed of water and methanol with gradient elution system, and flow rate is 1.0 ml/min. The detection wavelength was set at 205 nm.</P><P><B>Results:</B></P><P>Validation of this analytical method was evaluated by linearity, precision, and accuracy test. This established method had good linearity (<I>R</I><SUP>2</SUP> > 0.997). The relative standard deviation values of intra- and inter-day testing were indicated that <2%, and accuracy is 91.66%–103.31% at intraday and 91.69%–103.31% at intraday. The results of recovery test were 92.60%–108.99%.</P><P><B>Conclusion:</B></P><P>In these results, developed method was accurate and reliable to the quality evaluation of campesterol, emodin 8-O-β-D-glucopyranoside, quercetin, and isoquercitrin isolated from <I>R. sachalinensis</I>.</P><P><B>SUMMARY</B></P><P><P>We have developed high-performance liquid analysis method for simultaneous determination of 4 compounds of <I>Reynoutria sachalinensis</I>.</P></P> >[FIG OMISSION]</BR><P><B>Abbreviations used:</B> HPLC: High-performance liquid chromatography, DAD: Diode array detector, LOD: Limit of detection, LOQ: Limit of quantitation, ICH: International Conference on Harmonisation.</P>

Quantitative analysis of the eight major compounds in the Samsoeum using a high-performance liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometer

Weon, Jin Bae,Yang, Hye Jin,Lee, Bohyoung,Ma, Jin Yeul,Ma, Choong Je Medknow PublicationsMedia Pvt Ltd 2015 Pharmacognosy magazine Vol.11 No.42

<P><B>Background:</B></P><P>Samsoeum was traditionally used for treatment of a respiratory disease.</P><P><B>Objective:</B></P><P>The simultaneous determination of eight major compounds, ginsenoside Rg3, caffeic acid, puerarin, costunolide, hesperidin, naringin, glycyrrhizin, and 6-gingerol in the Samsoeum using a high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD) and an electrospray ionization mass spectrometer was developed for an accurate and reliable quality assessment.</P><P><B>Materials and Methods:</B></P><P>Eight compounds were qualitative identified based on their mass spectra and by comparing with standard compounds and quantitative analyzed by HPLC-DAD. Separation of eight compounds was carried out on a LUNA C<SUB>18</SUB> column (S-5 μm, 4.6 mm i.d. ×250 mm) with gradient elution composed of acetonitrile and 0.1% trifluoroacetic acid.</P><P><B>Results:</B></P><P>The data showed good linearity (<I>R</I><SUP>2</SUP> > 0.9996). The limits of detection and the limits of quantification were <0.53 μg and 1.62 μg, respectively. Inter- and Intra-day precisions (expressed as relative standard deviation values) were within 1.94% and 1.91%, respectively. The recovery of the method was in the range of 94.24–107.90%.</P><P><B>Conclusion:</B></P><P>The established method is effective and could be applied to quality control of Samsoeum.</P>

고속역류크로마토그래피 기술을 이용한 생리활성 물질의 분리 및 정제

정동수(Dong Su Jung),신현재(Hyun-Jae Shin) 한국생물공학회 2010 KSBB Journal Vol.25 No.3

역류크로마토그래피 (counter-current chromatography, CCC)는 일련의 분배과정을 한개의 튜브 내에서 연속적으로 일어나도록 고안된 시스템으로서 컬럼으로는 polytetrafluoroethylene (PTFE) 튜브가 다층으로 감겨있는 원통형의 홀더 3개가 서로 기어를 통해 물려있으며, 홀더가 회전과 공전을 통해 튜브의 꼬임을 방지하는 rotary seal-free flow centrifuge 시스템으로 되어있다. 역상 HPLC (reverse phase HPLC)에서는 고정상이 실리카에 결합된 유기물단 (organic moiety)이 수용성 이동상 물질에 의해서 용매화 (solvated)되는 반면 CCC는 실리카 대신에 강한 중력장에 의해 분리되는 자유로운 용매가 고정상이 되며 이 고정상의 부피비율은 20-30%에 이른다. 즉 고체담체에 결합된 유기관능기 대신에 물과 섞이지 않는 hexane 같은 유기용제가 고정상으로 사용되는 것이다. 고속역류크로마토그래피 (high-performance countercurrent chromatography, HPCCC)는 CCC의 기능을 향상시킨 분리시스템으로서 높은 중력장하에서 높은 이동상 속도와 높은 분리효율과 짧은 분리시간을 특징으로 하고 있다. 특히 mg 단위에서 kg 단위로의 스케일업이 선형적으로 가능하다는 큰 장점을 지니고 있다. 이 총설에서는 현재까지 개발된 CCC의 일반적인 이론을 간략히 정리하고 최신 HPCCC 장비의 적용 예를 살펴보고 그 응용분야로서 생리활성물질의 분리 및 정제와 관련된 연구동향을 정리하였다. Many successive liquid-liquid extractions occur enabling purification of the crude material to occur. In high performance counter-current chromatography (HPCCC), crude material is partitioned between two immiscible layers of solvent phases. The stationary phase (SP) is retained by hydrodynamic force field effect and the mobile phase (MP) is pumped through the column. Purification occurs because of the different solubility of the components in the liquid mobile and stationary phases. There are many key benefits of liquid stationary phases such as high mass and volume injection loadings, total sample recovery, and easy scale-up. Many researchers showed that predictable scale-up from simple test is feasible with knowledge of the stationary phase retention for the planned process scale run. In this review we review the recent advances in HPCCC research and also describe the key applications such as natural products and synthetics (small or large molecules).

High-Performance Liquid Chromatography Analysis of Capsaicin Content in 16 Capsicum Fruits from Nepal

Bindu Thapa,Natasa Skalko-Basnet,Akihito Takano,Kazuo Masuda,Purusotam Basnet1 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.4

Capsicum fruit, a popular spice as chili pepper, is an important ingredient of the formulations used in traditional medicines. Moreover, Capsicum fruit is listed as an official drug in several pharmacopoeias. Capsaicin, the most abundant component in Capsicum fruit, exhibits its therapeutic and adverse effects in a dose-dependent manner. Therefore, the known capsaicin content is the prerequisite for optimizing any formulation based on Capsicum fruit as a crude drug. We studied 16 samples of Capsicum fruits grown at different altitudes in Nepal and determined their capsaicin content by high-performance liquid chromatography. The capsaicin content was found to range from 2.19 to 19.73mg/g of dry weight of Capsicum fruits. Capsaicin content in pericarp was found to be higher than in seeds. No correlation was found between the shape or size of the fruits and its capsaicin content. Our findings indicate that many of the formulations prepared from Capsicum fruit, even as described in pharmacopoeias, may vary in their strength, therapeutic activity, and possible side effects if the capsaicin content in Capsicum fruit is not standardized.

Quantification of maltol in Korean ginseng ( <i>Panax ginseng</i> ) products by high-performance liquid chromatography-diode array detector

Jeong, Hyun Cheol,Hong, Hee-Do,Kim, Young-Chan,Rhee, Young Kyoung,Choi, Sang Yoon,Kim, Kyung-Tack,Kim, Sung Soo,Lee, Young-Chul,Cho, Chang-Won Medknow PublicationsMedia Pvt Ltd 2015 Pharmacognosy magazine Vol.11 No.43

<P><B>Background:</B></P><P>Maltol, as a type of phenolic compounds, is produced by the browning reaction during the high-temperature treatment of ginseng. Thus, maltol can be used as a marker for the quality control of various ginseng products manufactured by high-temperature treatment including red ginseng. For the quantification of maltol in Korean ginseng products, an effective high-performance liquid chromatography-diode array detector (HPLC-DAD) method was developed.</P><P><B>Materials and Methods:</B></P><P>The HPLC-DAD method for maltol quantification coupled with a liquid-liquid extraction (LLE) method was developed and validated in terms of linearity, precision, and accuracy. An HPLC separation was performed on a C18 column.</P><P><B>Results:</B></P><P>The LLE methods and HPLC running conditions for maltol quantification were optimized. The calibration curve of the maltol exhibited good linearity (<I>R</I><SUP>2</SUP> = 1.00). The limit of detection value of maltol was 0.26 μg/mL, and the limit of quantification value was 0.79 μg/mL. The relative standard deviations (RSDs) of the data of the intra- and inter-day experiments were <1.27% and 0.61%, respectively. The results of the recovery test were 101.35–101.75% with an RSD value of 0.21–1.65%. The developed method was applied successfully to quantify the maltol in three ginseng products manufactured by different methods.</P><P><B>Conclusion:</B></P><P>The results of validation demonstrated that the proposed HPLC-DAD method was useful for the quantification of maltol in various ginseng products.</P>

High Performance Liquid Chromatography를 이용한 실리콘 하이드로겔렌즈의 지방 침전물 분석

강유선,이군자 대한시과학회 2010 대한시과학회지 Vol.12 No.2

목적: In vitro 조건 하에서 재질이 다른 실리콘 하이드로겔 렌즈에 결합되는 눈물의 지방성분인 올레산(oleic acid), 올레산에스테르(oleic acid methyl ester) 및 콜레스테롤(cholesterol)을 각각 정량하여 실리콘 하이드로겔렌즈에 부착되는 지방침전물을 비교하였다. 방법: Lotrafilcon A, lotrafilcon B, galyficon A 및 balafilcon A 재질의 실리콘 하이드로겔 렌즈와polymacon 렌즈 및 RGP 렌즈를 올레산, 올레산에스테르 및 콜레스테롤을 동일한 양 포함하는 식염수 용액에 담가 37℃에서 흔들어주면서 24시간 동안 오염시킨 후, methanol과 chloroform을 1:1(v/v) 비율로 포함하는 유기용매를 이용하여 4시간 동안 렌즈에 부착된 지방을 추출하였다. 추출된 지방은 C-18 column을 사용하여 HPLC(high performance liquid chromatography)를 수행하여 분리하고 정량하였다. 결과: 실리콘 하이드로겔렌즈에 부착된 지방의 총량은 재질의 종류에 따라 48~67 ㎍/lens, polymacon 렌즈와 RGP 렌즈에서는 각각 18 ㎍/lens, 177 ㎍/lens로 측정되어, 실리콘 하이드로겔렌즈의 지방 부착량은 polymacon 소프트렌즈의 300~372%, RGP 렌즈의 30.5~37.8%로 나타났다. 지방침전물은 galyfilcon A 재질에 가장 많이 부착되었으며(p Purposes: The purpose of this study is to investigate and compare the in vitro sorption of major tear film lipids such as oleic acid, oleic acid methyl ester and cholesterol on the silicone hydrogel materials. Methods: Lipid spoilation was performed by incubating a lotrafilcon A, lotrafilcon B, galyfilcon A and balafilcon A silicon hydrogel lenses, polymacon soft contact lenses and RGP lenses in phosphate buffered saline (PBS) containing oleic acid, oleic acid methyl ester, and cholesterol at 37℃ for 24 hours with shaking. Lipids were extracted from the lenses with a solvent containing 1:1 methanol/chloroform (v/v) for 4 hours. High performance liquid chromatography methods using C-18 column were used to analyze lipid deposit extracts from contact lenses. Results: The amount of total lipids on silicone hydrogel contact lenses were 48~67 ㎍per lens depending on the silicone hydrogel lens materials versus 18 and 177 ㎍per lens for polymacon and RGP contact lenses, respectively that showed 300~372% and 30.5~37.8% of that of polymacon soft contact lenses and RGP lenses, respectively. After 24 hours of soaking, galyfilcon A showed the highest adsorption of all the silicone hydrogel lenses (p

High Performance Liquid Chromatography에 의한 마황 분획물의 ephedrine분석

조여원,임경아 경희대학교 생활과학연구소 2000 생활과학논집 Vol.4 No.1

Ma-huang is a traditional Chinese medicinal herb derived from Ephedra sinica Stapf. and other Ephedra species used to treat asthma, nose and lung congestion, and fever with anhidrosis. It contains 0.5-2.5% by weigh of total alkaloids, of which ephedrine accounts for 30-90%. Recently, large amounts of ma-huang were used as a source of ephedrine in many dietary supplements formulated for anti-obesity, because ephedrine has been found to be effective on inducing weight loss for obese patients. Ephedrine in Ephedrac Herba extracts was analyzed by high performance liquid chromatography(HPLC). The HPLC system(HP series 1100) consisted of quatemary pump, a degasser, a Diode Array Detection(DAD) system and HP Chem-Section for data analysis. Separation was achieved on a Nova-Pak C_18 column(150*3.9mm ID). The mobile phase consisted of methanol : water : acetic acied = 35:65:0.5 and flow rate was 1.0ml/min. The mobile phase was filtered through a millipore vacuum filter system equipped with a 0.45μm filter before use. The UV detector was set at 290nm and the column temperature was 30˚C. The injection volume of the samples was 10μl. The purity of the chromatographic peaks was assessed by suitable algorithm of the detector software. Calculations were performed by the external standard method. The results are summarized as follows. The percent yield of Ephedrac Herba extracts 199g for 85% methanol and its subfraction, hexane, ethylacetate, butanol, aqueous fraction and heat-water were 15.99%, 0.87%, 0.89%, 1.80%, 2.68% and 12.71%. The yield weight of the ephedrine extracts fo 100g of Ephedrae herna for methanol, hexane, ethylacetate, butanol,, aqueous fraction, and heat-water extract were 3.65mg, 0.24mg, 0.29mg, 0.12mg, 0.02mg and 9.49mg, respectively.

Simultaneous and rapid analysis of chemical preservatives in processed animal products by ultra-performance liquid chromatography

Park, Jwahaeng,Choi, Sunju,Oh, Donghwan,Mah, Jae-Hyung 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.1

An ultra-performance liquid chromatography-tunable ultraviolet method was optimized and validated for the simultaneous analysis of nine chemical preservatives in processed animal products. The limits of detection and quantification for the preservatives were within the ranges of 0.02-0.23 and $0.07-0.76{\mu}g/mL$, respectively. The relative standard deviations for intraday analyses of retention time and peak area were 0.00-0.23 and 0.03-2.93%, respectively, whereas, those for interday analyses were 0.67-2.30 and 2.12-5.37%, respectively. Of the nine preservatives spiked into six different animal products, dehydroacetic acid spiked into soft cheese exhibited the lowest recovery rate of $72.1{\pm}0.36%$ at the lowest concentration (0.25 g/kg). Comparing data between UPLC and high-performance liquid chromatography with a 5% significance level, the t-statistic was 1.42. Moreover, sorbic acid was detected in 16 animal products (0.11-2.49 g/kg) when 278 products were analyzed for preservatives.

Simultaneous and rapid analysis of chemical preservatives in processed animal products by ultra-performance liquid chromatography

박좌행,최순주,오덕환,마재형 한국식품과학회 2018 Food Science and Biotechnology Vol.27 No.1

An ultra-performance liquid chromatographytunable ultraviolet method was optimized and validated for the simultaneous analysis of nine chemical preservatives in processed animal products. The limits of detection and quantification for the preservatives were within the ranges of 0.02–0.23 and 0.07–0.76 lg/mL, respectively. The relative standard deviations for intraday analyses of retention time and peak area were 0.00–0.23 and 0.03–2.93%, respectively, whereas, those for interday analyses were 0.67–2.30 and 2.12–5.37%, respectively. Of the nine preservatives spiked into six different animal products, dehydroacetic acid spiked into soft cheese exhibited the lowest recovery rate of 72.1 ± 0.36% at the lowest concentration (0.25 g/kg). Comparing data between UPLC and high-performance liquid chromatography with a 5% significance level, the t-statistic was 1.42. Moreover, sorbic acid was detected in 16 animal products (0.11–2.49 g/kg) when 278 products were analyzed for preservatives.

Simultaneous Determination of Four Marker Components in Yukmijihwang Tang by High Performance Liquid Chromatography/Diode Array Detector

Won, Jin-Bae,Ma, Jin-Yeul,Ma, Choong-Je 대한약학회 2010 Archives of Pharmacal Research Vol.33 No.4

Simultaneous determination method of four marker components, paeoniflorin, loganin, 5-hydroxymethyl-2-furaldehyde and paeonol in traditional herbal medicine, Yukmijihwang tang, was developed to achieve rapidly and systematic quality control by high performance liquid chromatography coupled with diode array detection. To develop this high performance liquid chromatography method, $C_{18}$ column ($5\;{\mu}m$, $120\;{\AA}$, $4.6\;mm{\times}150\;mm$) was used with gradient elution of water and methanol as mobile phase. Validation of the chromatography method was evaluated by linearity, recovery, and precision test. Calibration curves of standard components showed excellent linearity with $R^2$ > 0.9990. Limits of detection and limits of quantification were in the ranges 0.01-0.02 and $0.04-0.07\;{\mu}g$/mL, respectively. The relative standard deviations of data of the intra-day and inter-day experiments were less than 1.15% and 3.76%, respectively. The results of recovery test were found to range from 94.88 to 107.43% with relative standard deviation values 0.07-2.66%. The results of validation suggested that this method was very accurate and stabilized.