• Natural Product Sciences
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• 제25권4호
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• pp.334-340
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• 2019

Cuscuta chinensis Lam. and Cuscuta japonica Choisy are parasitic plants. C. chinensis seeds were traditionally used for treatment of kidney and liver deficiencies. C. japonica seeds were used as tonic medicine to improve liver function and strengthen kidneys, treatment of high blood pressure, chronic diarrhea, and sore eyes. Cuscutae Semen are seeds of only C. chinensis in Korean Herbal Pharmacopoeia (K.H.P.). The developed HPLC-PDA method easily, accurately, and sensitively quantified using eight marker compounds [hyperoside (1), astragalin, (2), quercetin (3), kaempferol (4), chlorogenic acid (5), 3,4-di-O-caffeoylquinic acid (6), 1,5-di-O-caffeoylquinic acid (7), and 4,5-di-O-caffeoylquinic acid (8)]. In addition, the method may be used to distinguish seeds between C. chinensis Lam. and C. japonica Choisy. Furthermore, the result from the current study was applied to clarify samples between steam processed and unprocessed samples of C. chinensis by pattern analysis.

• 한국산학기술학회논문지
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• 제18권12호
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• pp.755-761
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• 2017

푸코잔틴을 함유한 원료 및 이를 유효성분으로 처방된 기능성 화장품의 품질관리를 위하여 photodiode array (PDA) 검출기를 갖춘 HPLC를 이용한 분석법을 확립하고 원료 및 제품 함량 분석에 응용하고자 본 연구를 수행하였다. 컬럼은 옥타데실화한 실리카 겔을 충진한 것을 사용하였으며 PDA의 측정파장은 499 nm로 설정하였다. 분석법을 밸리데이션하기 위하여 검량선의 직선성, 검출 및 정량 한계, 재현성, 회수율을 조사한 결과 양호한 결과를 얻었다. 검량선의 상관계수는 1.000로 0.5 ~ 100 ppm의 농도에서 직선성이 양호하였다. 또한 검출 한계는 0.1 ppm이었으며 정량 한계는 0.5 ppm이었다. 시스템의 적합성을 확인하기 위하여 피크의 재현성을 평가한 결과 피크 면적 값의 RSD (n=5) 값은 2.0%이었으며 머무름 시간의 RSD는 0.09% 이었다. 표준물질 첨가법의 실험 결과 회수율은 $101.6{\pm}0.77%$이었다. 끝으로 확립된 분석법을 원료 및 제품 분석에 적용하였다. 두 종류의 푸코잔틴 함유 원료의 푸코잔틴 함량은 각각 $49.6{\pm}3.3%$와 $1.03{\pm}0.016%$이었다. 또한 푸코잔틴이 150 ppm이 되도록 처방한 제품의 푸코잔틴의 함량은 $156.7{\pm}4.7ppm$으로 확인되었다. 위 실험결과로 볼 때 본 실험방법은 기능성 화장품에서 푸코잔틴을 정량분석하는 것에 적용될 수 있을 것으로 판단되었다.

• 대한한의학방제학회지
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• 제18권1호
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• pp.95-103
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• 2010

Objectives : To develop and validate HPLC-PDA methods for simultaneous determination of seven constituents in Samsoeum(SSE). Methods : Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array(PDA) detection at 254 and 280 nm, were used for quantification of the seven marker components of SSE. 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. Results : Calibration curves were acquired with $r^2$>0.9997, and the relative standard deviation (RSD) values (%) for intra- and inter-day precision were less than 3.0%. The recovery rate of each compound was in the range of 100.07-112.65%, with an RSD less than 4.0%. The contents of seven compounds in SSE were 1.24-10.53 mg/g. Conclusions : The established method will be helpful to improve quality control of SSE.

• 대한본초학회지
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• 제25권3호
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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.

• Natural Product Sciences
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• 제25권2호
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• pp.122-129
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• 2019

The roots of Phlomis umbrosa (Turcz.) (Phlomidis Radix) have been traditionally used to treat cold, reduce swelling and staunch bleeding. Four iridoids (1 - 3 and 5) and six phenylethanoid derivatives (4, and 6 - 10) were isolated from the roots of P. umbrosa. A simple, sensitive, and reliable analytical HPLC/PDA method was developed, validated, and applied to determine 10 marker compounds in Phlomidis Radix. Furthermore, the isolates were evaluated for cytotoxic and anti-oxidant activities as well as DPPH-HPLC method. Among them, compounds 4 and 6 - 9 displayed potent anti-oxidant capacities using DPPH assay with $IC_{50}$ values of $27.7{\pm}2.4$, $10.2{\pm}1.1$, $18.0{\pm}0.8$, $19.1{\pm}0.3$, and $19.9{\pm}0.6{\mu}M$, and compounds 6, 8, and 9 displayed significant cytotoxic activity against HL-60 with $IC_{50}$ values of $35.4{\pm}3.1$, $18.6{\pm}2.0$, and $42.9{\pm}3.0{\mu}M$, respectively.

• 한국식품영양과학회지
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• 제46권2호
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• pp.210-219
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• 2017

손바닥선인장으로부터 기능성식품 개발을 위하여 최적의 에탄올 추출물 탐색과 HPLC-PDA 분석방법에 의한 validation을 실시하였다. 지표성분으로 dihydrokaempferol (DHK)과 3-O-methylquercetin(3-MeQ)을 선정하여 표준화를 실시하였으며 검출법 확립을 위한 정량분석은 Luna RP-18 칼럼($4.6{\times}250mm$, $5{\mu}m$)을 이용하여 1% 인산용액과 아세토니트릴을 전개 용매로 사용하였다. 용출은 1.0 mL/min의 유속으로 기울기 용출(gradient elution) 방법을 이용하였으며, 280 nm 파장에서는 DHK를, 360 nm 파장에서는 3-MeQ를 검출한 피크 면적을 이용하여 검량곡선을 작성하여 분석하였다. 본 연구에서 확립한 분석법으로 특이성, 직선성, 정밀성, 정확성, 회수율을 검색하였다. DHK의 검량선으로부터 상관계수($R^2$) 0.9998의 우수한 직선성과 intra-day와 inter-day 분석에서 97% 이상의 회수율과 3% 미만의 RSD를 나타내 정밀성과 정확성을 입증하였다. 3-MeQ의 검량선으로부터 상관계수($R^2$) 1의 우수한 직선성과 intra-day와 inter-day 분석에서 95% 이상의 회수율과 7% 미만의 RSD를 나타내 정밀성과 정확성을 입증하였다. DHK의 검출한계는 $1.38{\mu}g/mL$, 정량한계는 $4.18{\mu}g/mL$로 나타났으며, 3-MeQ의 검출한계는 $3.49{\mu}g/mL$, 정량한계는 $10.6{\mu}g/mL$로 나타났다. 손바닥선인장 에탄올 추출물(OFSEs)은 70과 $80^{\circ}C$에서 50, 70, 80% 에탄올로 3, 4, 5, 6시간 동안 각각 추출하였으며, 지표물질의 검량곡선을 활용하여 각각의 OFSEs로부터 두 종의 지표물질 함량을 분석하였다. 본 시험법으로 분석한 지표물질의 함량은 $80^{\circ}C$에서 추출한 70% OFSE가 DHK $26.42{\pm}0.65mg/OFS100g$, 3-MeQ $3.88{\pm}0.29mg/OFS100g$의 함량을 나타내 가장 우수하게 나타났다. 다양한 OFSEs에 대하여 DPPH 자유 라디칼 소거효능과 쥐의 간 균질액을 이용한 지질과산화 저해 효능에 대한 항산화 효능은 지표물질의 함량이 가장 높은 70% OFSE에서 우수한 효능을 나타내 본 연구에서 확립한 원료 표준화를 위한 적합한 분석법임이 검증되었다. 따라서 본 연구를 통하여 HPLC-PDA를 이용한 손바닥선인장 에탄올 추출물의 DHK와 3-MeQ의 분석법은 개별인정형 건강 기능식품 기능성 원료 개발을 위한 유용한 자료로 활용될 것으로 생각한다.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.67-72
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• 2023

A simple and accurate method was developed for the quantitative analysis of oleanonic acid (OA) from mastic gum. The analysis was carried out using reverse-phase high-performance liquid chromatography combined with a photodiode array detector (HPLC/PDA). Our optimized method was validated by measuring various parameters, using an INNO C18 column fitted with a gradient elution system. The results revealed limits of detection and quantification of 0.34 and 1.042 ㎍/mL, respectively. The OA calibration curve exhibited excellent linearity over the concentration range of 0.0625 to 2.0 mg/mL, with r² =0.9996. Accuracy tests revealed a high recovery rate of 99.44-103.66%, with precision values below 0.15%. These results suggest that the present analytical method can identify and quantify OA in mastic gum with high precision. The HPLC approach developed in this study might be applied to routine analyses and large-scale extraction procedures for OA content quantification.

• 대한한의학회지
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• 제31권6호
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• pp.8-15
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• 2010

Objectives: We investigated to develop and validate HPLC-PDA methods for simultaneous determination of eight constituents in Galgeun-tang (GGT). Methods: Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 230 nm, 254 nm, and 280 nm, were used for quantification of the eight marker components of GGT. 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. 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 3.0%. The recovery rate of each component was in the range of 87.33-101.38%, with an RSD less than 7.0%. The contents of the eight components in GGT were 1.98-12.17 mg/g. Conclusions: The established method will be applied for the quantification of marker components in GGT.

• Natural Product Sciences
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• 제17권4호
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• pp.337-341
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• 2011

An HPLC (high-performance liquid chromatography)-PDA (photodiode array) detection method was established for the determination of naringin, hesperidin and neohesperidin in the fruit of Citrus paradisi and C. grandis. The flavonoids were separated in less than 20 min using an YMC RP 18 column with isocratic elution using acetonitrile and water (23 : 77, v/v) at a flow rate of 1 ml/min, and a PDA detector. The levels of naringin, hesperidin and neohesperidin were 1345.92, 950.62, and 2078.82 ${\mu}g/g$, respectively, in the peel, and 102.43, 59.13, and 86.68 ${\mu}g/g$, respectively, in the flesh of C. paradisi. In C. grandis, the levels of naringin, hesperidin and neohesperidin were 3530.56, 80.00, and 5.26 ${\mu}g/g$, respectively, in the peel, and 59.59, 7.43, and 38.41 ${\mu}g/g$, respectively, in the flesh. The total content was highest in the peel, reaching 0.44% and 0.36% in C. paradisi and C. grandis, respectively, while the flesh contained only 0.025% and 0.011%, respectively. Therefore, the peels of C. paradisi and C. grandis are necessary for the processing and utilization of flavonoids.

• 대한한의학방제학회지
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• 제21권1호
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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.