Malaysian Journal of Analytical Sciences Vol 19 No 3 (2015): 557 – 564

 

 

 

SPECTROFLUORIMETRIC DETERMINATION OF BRASSINOSTEROIDS PLANT HORMONES IN BIO-EXTRACT SAMPLES

 

(Penentuan spektrofluorimetrik bagi Hormon Tumbuhan Brassinosteroids dalam Sampel

Bio-Ekstrak)

 

Napa Tangtreamjitmun* and Kanokporn Chindaphan

 

Department of Chemistry,

Faculty of Science,

Burapha University, Mueang, Chonburi, 20131 Thailand

 

*Corresponding author: napa@buu.ac.th

 

 

Received: 13 April 2015; Accepted: 19 May 2015

 

 

Abstract

A spectrofluorometric method was developed for determination of brassinosteroids plant hormones based on the reaction between brassinolide and m-dansylaminophenylboronic acid. 90 µM of m-dansylaminophenylboronic acid solution was chosen as the optimal concentration of the reagent. The temperature of the reaction was 30°C (room temperature) for 20 min duration in 5 mM phosphate buffer solution, pH 7. The excitation and emission wavelengths were 380 nm and 505 nm, respectively. Under optimal condition, linearity was found in the range 1-50 µM. Standard addition was used for the brassinosteroids determination in bio-extract samples, without any sample preparation. Limit of detection (3SD, n=10) of 0.24 µM and limit of quantitation (10SD, n=10) of 0.48 µM were obtained using sample blank with this standard addition method. Result was demonstrated to be a suitable semi-quantitative technique and was applied successfully for the determination of total brassinosteroids plant hormones equivalent to brassinolide. 

 

Keywords: brassinolide, brassinosteroids, m-dansylaminophenylboronic acid, bio-extract, spectrofluorometry

 

Abstrak

Kaedah spektroflourometrik telah dibangunkan bagi penentuan hormon tumbuhan brassinosteroids berdasarkan tindak balas antara brassinolide dan asid m-dansylaminofenilboronik. 90 μM larutan asid m-dansylaminofenilboronik dipilih sebagai reagen pada kepekatan optimum. Suhu tindak balas adalah 30 °C (suhu bilik) untuk tempoh 20 min di dalam 5 mM larutan penimbal fosfat, pH 7. Panjang gelombang pengujaan dan pancaran masing – masing adalah 380 nm dan 505 nm. Di bawah keadaan optimum, kelinearan diperolehi di dalam julat 1-50 μM. Penambahan larutan piawai digunakan bagi penentuan brassinosteroids di dalam sampel bio-ekstrak, tanpa sebarang penyediaan sampel. Had pengesanan (3SD, n = 10) iaitu 0.24 μM dan had kuatifikasi (10SD, n = 10) iaitu 0.48 μM telah diperolehi dengan menggunakan sampel pengosong dan larutan piawai yang ditambah. Keputusan menunjukkan bahawa kaedah semi kuantitatif ini sesuai dan boleh diaplikasikan bagi penentuan jumlah hormon brassinosteroids di dalam tumbuhan yang bersamaan dengan kandungan brassinolide.

 

Kata kunci: brassinolide, brassinosteroids, asid m-dansylaminofenilboronik, bio-ekstrak, spektroflourometri

 

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