Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 2, February 2008, pp. 150-156
Simultaneous Determination of Catechins, Rutin,
and Gallic Acid in Cistus Species Extracts by HPLC with Diode
Array Detection
Natale Alfredo Santagati[1], Loredana Salerno[1],
Giuseppina Attaguile[2], Francesca Savoca[3], and Giuseppe Ronsisvalle[1]
[1]Dipartimento di Scienze Farmaceutiche, Università di
Catania. Viale A. Doria 6, 95125 Catania, Italy;
[2]Dipartimento
di Farmacologia Clinica Sperimentale, Università di Catania.
Viale A. Doria 6, 95125 Catania, Italy; and
[3]Dipartimento di
Botanica, Università di Catania. Via A. Longo 19, 95100
Catania, Italy
A simple high-performance liquid chromatography
method using a diode array detector (DAD) is developed for the
simultaneous analysis of five major catechins: (+)-catechin (C),
(–)-epicatechin (EC), (–)-gallocatechin (GCT), (–)-epigallocatechin
(EGC), (–)-epigallocatechin gallate (EGCG), and the phenolic
plant metabolites gallic acid (GA) and rutin (RT) in lyophilized
extracts of Cistus species. The optimal analytical conditions
are investigated to obtain the best resolution and the highest
UV sensitivity for the quantitative detection of catechins. The
optimized conditions (acetonitrile–phosphate buffer 50mM,
pH 2.5, gradient elution system on a C18 reversed-phase column
with a flow rate of 1 mL/min and UV absorbance at 210 nm) allowed
a specific and repeatable separation of the studied analytes
to be achieved. All compounds are successfully separated within
32 min. Calibration curves are linear in the 2–50 µg/mL
range for GCT, C, and EGCG and in the 5–50 µg/mL
range for GA, EGC, EC, and RT. The limit of detection values
ranged from 0.24 to 0.74 µg/mL. The limit of quantitation
limit values ranged from 0.77 to 1.94 µg/mL. The validated
method is applied to the determination of the specific phytochemical
markers GA, GCT, C, and RT in Cistus incanus and Cistus monspeliensis
lyophilised extracts. The recovery values ranged between 78.7%
and 98.2%. The described HPLC method appears suitable for the
differentiation and determination of the most common catechins
together with the glycoside rutin and the phenolic compound gallic
acid and can be considered an effective and alternative procedure
for the analyses of this important class of natural compounds.
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