Abstract：
BACKGROUND: Cassia (Family: Fabaceae) species are a large group of flowering plants rich in bioactive anthraquinone and flavonoids used in botanical supplements and nutraceuticals.
OBJECTIVE: A simple and reliable high-performance liquid chromatography-photodiode array (HPLC-PDA) method was developed and validated for separating and quantifying 13 anthraquinone and flavonoids. These compounds were further confirmed using an LC-based electrospray ionization mass spectrometry (ESI-MS/MS) method in the leaves and flowers of selected Cassia species. A simple and rapid HPTLC method was developed for chemical fingerprint analysis of all Cassia species.
METHODS: All 13 compounds were chromatographically separated on a Zorbax TC18 (4.6 × 250, 5 μm particle size) analytical column, and 0.1% formic acid and acetonitrile as elution solvents at a flow rate of 0.8 mL/min with detection at 259 nm. For HPTLC fingerprinting, the mobile phase compositions of toluene, ethyl acetate, and formic acid (5.5:4.2:0.6, v/v/v) were optimized to separate and identify all compounds using silica gel 60F254 aluminum plates.
RESULTS: The validation data for the developed HPLC-PDA method for 13 compounds showed good linearity (r2 >0.99) with a sensitive LOD (0.082-1.969 μg/mL), LOQ (0.250-5.967 μg/mL), and excellent recoveries (85.22-100.32%). The quantification results were found to be precise and accurate (<5.0% and relative error), -0.77-0.44 with ESI-MS/MS confirmation in the Cassia samples. The novel HPTLC method was excellent separation for 13 compounds, with Rf values ranging between 0.12 and 0.61.
CONCLUSIONS: The developed HPLC-PDA method was simple and precise and could separate and quantify anthraquinones and flavonoids along with confirmation, using a novel LC-based ESI-MS/MS. The HPTLC method was found to be simple and precise, with excellent separation capabilities for these compounds.
CONCLUSIONS: This novel multiplatform approach successfully identified and quantified 13 compounds simultaneously using an integration of data strategy in seven medicinally important Cassia species\' leaves and flowers.
OBJECTIVE: A simple and reliable high-performance liquid chromatography-photodiode array (HPLC-PDA) method was developed and validated for separating and quantifying 13 anthraquinone and flavonoids. These compounds were further confirmed using an LC-based electrospray ionization mass spectrometry (ESI-MS/MS) method in the leaves and flowers of selected Cassia species. A simple and rapid HPTLC method was developed for chemical fingerprint analysis of all Cassia species.
METHODS: All 13 compounds were chromatographically separated on a Zorbax TC18 (4.6 × 250, 5 μm particle size) analytical column, and 0.1% formic acid and acetonitrile as elution solvents at a flow rate of 0.8 mL/min with detection at 259 nm. For HPTLC fingerprinting, the mobile phase compositions of toluene, ethyl acetate, and formic acid (5.5:4.2:0.6, v/v/v) were optimized to separate and identify all compounds using silica gel 60F254 aluminum plates.
RESULTS: The validation data for the developed HPLC-PDA method for 13 compounds showed good linearity (r2 >0.99) with a sensitive LOD (0.082-1.969 μg/mL), LOQ (0.250-5.967 μg/mL), and excellent recoveries (85.22-100.32%). The quantification results were found to be precise and accurate (<5.0% and relative error), -0.77-0.44 with ESI-MS/MS confirmation in the Cassia samples. The novel HPTLC method was excellent separation for 13 compounds, with Rf values ranging between 0.12 and 0.61.
CONCLUSIONS: The developed HPLC-PDA method was simple and precise and could separate and quantify anthraquinones and flavonoids along with confirmation, using a novel LC-based ESI-MS/MS. The HPTLC method was found to be simple and precise, with excellent separation capabilities for these compounds.
CONCLUSIONS: This novel multiplatform approach successfully identified and quantified 13 compounds simultaneously using an integration of data strategy in seven medicinally important Cassia species\' leaves and flowers.
摘要：
背景：决明（家族：豆科）物种是一类富含生物活性蒽醌和类黄酮的开花植物，用于植物补充剂和营养食品。
目的：开发并验证了一种简单可靠的高效液相色谱-光电二极管阵列（HPLC-PDA）方法，用于分离和定量13种蒽醌和黄酮类化合物。使用基于LC的电喷雾电离质谱（ESI-MS/MS）方法在选定的决明子物种的叶和花中进一步确认了这些化合物。开发了一种简单快速的HPTLC方法，用于所有决明子的化学指纹图谱分析。
方法：在ZorbaxTC18（4.6x250，5μm粒径）分析柱上对所有13种化合物进行色谱分离，和0.1%甲酸和乙腈作为洗脱溶剂,流速为0.8mL/min,在259nm处检测。对于HPTLC指纹，甲苯的流动相组成，乙酸乙酯,和甲酸(5.5:4.2:0.6,v/v/v)被优化以使用硅胶60F254铝板分离和鉴定所有化合物。
结果：13种化合物的已开发HPLC-PDA方法的验证数据显示出良好的线性（r2>0.99），具有灵敏的LOD（0.082-1.969mg/mL），LOQ（0.250-5.967mg/mL），和优良的回收率（85.22-100.32%）。定量结果准确准确(<5.0%,相对误差),-0.77-0.44,在决明子样品中具有ESI-MS/MS确认。新的HPTLC方法对13种化合物具有良好的分离效果，Rf值范围在0.12-0.61之间。
结论：开发的HPLC-PDA方法简单，并且精确，可以分离和定量蒽醌和类黄酮以及确认，使用基于LC的新型ESI-MS/MS。发现HPTLC方法简单而精确，对这些化合物具有优异的分离能力。
结论：这种新颖的多平台方法成功地同时鉴定和定量了13种化合物，使用数据策略整合在7种重要的决明子叶和花中。
目的：开发并验证了一种简单可靠的高效液相色谱-光电二极管阵列（HPLC-PDA）方法，用于分离和定量13种蒽醌和黄酮类化合物。使用基于LC的电喷雾电离质谱（ESI-MS/MS）方法在选定的决明子物种的叶和花中进一步确认了这些化合物。开发了一种简单快速的HPTLC方法，用于所有决明子的化学指纹图谱分析。
方法：在ZorbaxTC18（4.6x250，5μm粒径）分析柱上对所有13种化合物进行色谱分离，和0.1%甲酸和乙腈作为洗脱溶剂,流速为0.8mL/min,在259nm处检测。对于HPTLC指纹，甲苯的流动相组成，乙酸乙酯,和甲酸(5.5:4.2:0.6,v/v/v)被优化以使用硅胶60F254铝板分离和鉴定所有化合物。
结果：13种化合物的已开发HPLC-PDA方法的验证数据显示出良好的线性（r2>0.99），具有灵敏的LOD（0.082-1.969mg/mL），LOQ（0.250-5.967mg/mL），和优良的回收率（85.22-100.32%）。定量结果准确准确(<5.0%,相对误差),-0.77-0.44,在决明子样品中具有ESI-MS/MS确认。新的HPTLC方法对13种化合物具有良好的分离效果，Rf值范围在0.12-0.61之间。
结论：开发的HPLC-PDA方法简单，并且精确，可以分离和定量蒽醌和类黄酮以及确认，使用基于LC的新型ESI-MS/MS。发现HPTLC方法简单而精确，对这些化合物具有优异的分离能力。
结论：这种新颖的多平台方法成功地同时鉴定和定量了13种化合物，使用数据策略整合在7种重要的决明子叶和花中。
