고성능 액체 크로마토그래피 (HPLC) 및 초고성능 액체 크로마토그래피 (UPLC)와 chaotropic mobile phase additive를 이용하여 현호색 (Corydalis Tuber)의 tertiary 및 quaternary isoquinoline alkaloid의 동시 ...
고성능 액체 크로마토그래피 (HPLC) 및 초고성능 액체 크로마토그래피 (UPLC)와 chaotropic mobile phase additive를 이용하여 현호색 (Corydalis Tuber)의 tertiary 및 quaternary isoquinoline alkaloid의 동시 분석법을 개발하였다. 4종의 tertiary alkaloid (glaucine, protopine, tetrahydropalmatine 및 corydaline)와 4종의 quaternary alkaloid (coptisine, palmatine, berberine 및 dehydrocorydaline), 총 8종의 isoquinoline alkaloid를 분석 대상 성분으로 하였다. HPLC 분석법의 최적 이동상으로 NaBF4 30 mmol/L를 첨가한 10 mmol/L potassium dihydrogen phosphate (pH 3.0, phosphoric acid) 완충액과 acetonitrile을 선정하였고, 최적 칼럼으로 fused-core particle로 충전된 Ascentis Express C18 (4.6 mm x 150 mm, 2.7 μm) 칼럼을 선정하였다. UPLC 분석법은 NaPF6 30 mmol/L를 첨가한 10 mmol/L potassium dihydrogen phosphate (pH 2.5, phosphoric acid) 완충액과 acetonitrile이 최적의 이동상으로, Waters Acquity UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm)을 최적 칼럼으로 선정하였다. 칼럼 온도는 25°C로 유지하였고 검출 파장은 280 nm를 사용하였다. 직선성, 정밀성, 정확성, 정량한계 (LOQ) 및 회수율을 평가하여 개발된 UPLC 분석법의 적합성, 신뢰성을 입증하였다. 확립된 UPLC 분석법을 적용하여 현호색 시료 30종에 함유된 8가지 isoquinoline alkaloid의 함량을 분석하였다. 다변량 통계 분석 방법으로 군집 분석 (cluster analysis)과 대응 분석 (correspondance analysis)을 사용하여 30종 현호색 시료의 패턴을 확인하였다. 본 연구 결과가 현호색의 품질관리 및 alkaloid 분석에 유용하게 활용될 수 있기를 기대한다.
고성능 액체 크로마토그래피 (HPLC) 및 초고성능 액체 크로마토그래피 (UPLC)와 chaotropic mobile phase additive를 이용하여 현호색 (Corydalis Tuber)의 tertiary 및 quaternary isoquinoline alkaloid의 동시 분석법을 개발하였다. 4종의 tertiary alkaloid (glaucine, protopine, tetrahydropalmatine 및 corydaline)와 4종의 quaternary alkaloid (coptisine, palmatine, berberine 및 dehydrocorydaline), 총 8종의 isoquinoline alkaloid를 분석 대상 성분으로 하였다. HPLC 분석법의 최적 이동상으로 NaBF4 30 mmol/L를 첨가한 10 mmol/L potassium dihydrogen phosphate (pH 3.0, phosphoric acid) 완충액과 acetonitrile을 선정하였고, 최적 칼럼으로 fused-core particle로 충전된 Ascentis Express C18 (4.6 mm x 150 mm, 2.7 μm) 칼럼을 선정하였다. UPLC 분석법은 NaPF6 30 mmol/L를 첨가한 10 mmol/L potassium dihydrogen phosphate (pH 2.5, phosphoric acid) 완충액과 acetonitrile이 최적의 이동상으로, Waters Acquity UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm)을 최적 칼럼으로 선정하였다. 칼럼 온도는 25°C로 유지하였고 검출 파장은 280 nm를 사용하였다. 직선성, 정밀성, 정확성, 정량한계 (LOQ) 및 회수율을 평가하여 개발된 UPLC 분석법의 적합성, 신뢰성을 입증하였다. 확립된 UPLC 분석법을 적용하여 현호색 시료 30종에 함유된 8가지 isoquinoline alkaloid의 함량을 분석하였다. 다변량 통계 분석 방법으로 군집 분석 (cluster analysis)과 대응 분석 (correspondance analysis)을 사용하여 30종 현호색 시료의 패턴을 확인하였다. 본 연구 결과가 현호색의 품질관리 및 alkaloid 분석에 유용하게 활용될 수 있기를 기대한다.
High performance liquid chromatography (HPLC) and ultra performance liquid chromatography (UPLC) methods using chaotropic mobile phase additives were developed to simultaneously determine tertiary and quaternary isoquinoline alkaloids derived from Corydalis Tuber. Analytical target compounds were ei...
High performance liquid chromatography (HPLC) and ultra performance liquid chromatography (UPLC) methods using chaotropic mobile phase additives were developed to simultaneously determine tertiary and quaternary isoquinoline alkaloids derived from Corydalis Tuber. Analytical target compounds were eight isoquinoline alkaloids, including four tertiary alkaloids (glaucine, protopine, tetrahydropalmatine and corydaline) and four quaternary alkaloids (coptisine, palmatine, berberine and dehydrocorydaline). The optimal column for HPLC method was Ascentis Express C18 (4.6 mm x 150 mm, 2.7 μm) column which was filled with the fused-core particles and the optimal mobile phase consisted of 10 mmol/L potassium dihydrogen phosphate in 30 mmol/L NaBF4 (sodium tetrafluoroborate) and acetonitrile. The optimal column for UPLC method was Waters Acquity UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm) column and the optimal mobile phase consisted of 10 mmol/L potassium dihydrogen phosphate in 30 mmol/L NaPF6 (sodium hexafluorophosphate) and acetonitrile. The column temperature was set at 25°C and the detection wavelength was 280 nm. The developed UPLC method was successfully validated in terms of linearity, precision, accuracy, limit of quantitation (LOQ) and recovery. This UPLC method was applied to determine eight isoquinoline alkaloids in 30 Corydalis Tuber samples and multivariate analysis was performed using correspondance analysis and cluster analysis to differentiate the 30 samples. These analytical methods could be valuable for the quality control of Corydalis Tuber and its pharmaceutical preparations.
High performance liquid chromatography (HPLC) and ultra performance liquid chromatography (UPLC) methods using chaotropic mobile phase additives were developed to simultaneously determine tertiary and quaternary isoquinoline alkaloids derived from Corydalis Tuber. Analytical target compounds were eight isoquinoline alkaloids, including four tertiary alkaloids (glaucine, protopine, tetrahydropalmatine and corydaline) and four quaternary alkaloids (coptisine, palmatine, berberine and dehydrocorydaline). The optimal column for HPLC method was Ascentis Express C18 (4.6 mm x 150 mm, 2.7 μm) column which was filled with the fused-core particles and the optimal mobile phase consisted of 10 mmol/L potassium dihydrogen phosphate in 30 mmol/L NaBF4 (sodium tetrafluoroborate) and acetonitrile. The optimal column for UPLC method was Waters Acquity UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm) column and the optimal mobile phase consisted of 10 mmol/L potassium dihydrogen phosphate in 30 mmol/L NaPF6 (sodium hexafluorophosphate) and acetonitrile. The column temperature was set at 25°C and the detection wavelength was 280 nm. The developed UPLC method was successfully validated in terms of linearity, precision, accuracy, limit of quantitation (LOQ) and recovery. This UPLC method was applied to determine eight isoquinoline alkaloids in 30 Corydalis Tuber samples and multivariate analysis was performed using correspondance analysis and cluster analysis to differentiate the 30 samples. These analytical methods could be valuable for the quality control of Corydalis Tuber and its pharmaceutical preparations.
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