Validation of Rutin Analysis in Cassava (Manihot esculenta) Leaves Using TLC-densitometer

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Pornarong Intarakasem
Somsak Nualkaew
Tanit Padumanonda


Introduction: Cassava leaf is the lesser used part of cassava compared to the root, which
is a well known source of tapioca fl our. Cassava leaves contain several compounds with health
benefi ts, especially a fl avonoid called rutin. The aim of this study was to develop and validate method
for quantitative analysis of rutin in cassava leaf extract. Methods: All cassava leaves were extracted
with 80% methanol and applied semi-automatically in TLC aluminum sheets, precoated with 0.2 mm
thick layer of silica gel 60 F-254. The mobile phases were ethyl acetate-acetone – water [40:50:10]
and ethyl acetate – formic acid- water [60:10:10]. The extracts were analyzed for rutin content using
TLC densitometry. Results: The solvent system gave a compact spot for rutin with Rf value of 0.47.
Linear regression analysis data for the calibration curve of rutin showed good linear relationship with
r2 of 0.999 in the concentration range from 200 – 1000 ng/spot. The method was validated for interday
and intraday precision (overall % RSD < 6.91 %) and accuracy (% recovery = 97.69 -99.96 %). The
limit of detection and quantifi cation were 20 ng/spot and 100 ng/spot, respectively. The developed
method was applied to determine rutin contents in cassava leaf extract from Kasetsart 50 cultivar and
the contents was 12.30±0.61% dry weight. Conclusions: The developed TLC-densitometric methods
provided a good accuracy and selectivity for the quantitative determination of rutin in cassava leaves.
The rutin content analyzed by this method is effective and reliable.

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