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Development and validation studies for determination of phenylpropanoid‐substituted flavan‐3‐ols in semipurified extract of T richilia catigua by high‐performance liquid chromatography with photodiode array detection
Author(s) -
Longhini Renata,
Klein Traudi,
Luciano Bruschi Marcos,
Silva Waldir Veríssimo,
Rodrigues Juliana,
Lopes Norberto Peporine,
Mello João Carlos Palazzo
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201200911
Subject(s) - chromatography , chemistry , repeatability , high performance liquid chromatography , detection limit , catechin , cinnamic acid , elution , polyphenol , organic chemistry , antioxidant
The phenolic compounds are the main phytochemical constituents of the bark of T richilia catigua and are commonly used for medicinal purposes. An HPLC method for the simultaneous quantification of phenolic compounds (procyanidin B2 ( PB 2), epicatechin ( EPC ), chinchonains Ia, Ib, IIa, IIb, catechin, and chrologenic acid) in T. catigua extract was developed and validated. A suitable chromatographic system was selected, which uses a gradient elution with methanol/ ACN (75:25), and water both with 0.05% TFA , as mobile phase, column Luna, 280 nm, and flow 0.4 mL/min. Validation of the analytical method was based on the parameters: linearity, precision, LOD s and LOQ s, accuracy, robustness, and stability. The method showed linearity for PB 2 and EPC , in the range 10–120 μg/mL with good correlation coefficients (>0.996). For precision, the repeatability ranged from 1.89 to 3.23%, and the values for accuracy for PB 2 and EPC were 95 and 89%, respectively. The LOD s and LOQ s for PB 2 were 1.36 and 4.12 μg/mL, and for EPC were 2.18 and 6.61 μg/mL, respectively. The method was robust under the conditions employed. The proposed method could be employed for quality assessment of T . catigua , as well as pharmaceutical products.