Malaysian Journal of Analytical Sciences Vol 19 No 3
(2015): 625 – 635
A SIMPLE THIN LAYER CHROMATOGRAPHY METHOD FOR SEPARATION OF SELECTED
NATURAL STEROID HORMONES
(Kaedah
Kromatografi Lapisan Nipis yang Mudah bagi Pemisahan Hormon Steroids Semulajadi
Terpilih)
*Corresponding author: kaja.kinga.rudnicka@gmail.com
Received: 14
April 2015; Accepted: 28 May 2015
Abstract
Chromatographic
properties of seven steroids: estrogens (β-estradiol
and estrone), androgens (testosterone, methyltestosterone, trans-androsterone),
progesterone and cholesterol have been studied by planar chromatography with
usage of High Performance Thin Layer Chromatography (HPTLC) and Thin Layer
Chromatography (TLC) plates. Normal, reversed and cyano-bonded silica
stationary phases were tested with five binary mobile phases (acetonitrile-water, acetonitrile-DMSO, acetonitrile-methanol,
acetone-petroleum ether, acetone-water) in which the concentration of organic
modifier varied from 0 to 100% (v/v).
This study reports the optimization of steroid hormones separation. Principal
Component Analysis (PCA) based on calculated molecular descriptors
quantitatively differentiating solutes was performed in order to investigate
the similarity and dissimilarity between tested compounds. The separation
abilities of mobile and stationary phases were compared based on separation factor
α. Chromatographic retention data and possible retention mechanisms also
were discussed.
Keywords: thin layer
chromatography, separation factor, binary phases, steroids
Abstrak
Sifat
kromatografi tujuh steroid: estrogen (β-estradiol dan estron), androgen
(testosteron, metiltestosteron, trans-androsteron), progesteron dan kolesterol
telah dikaji oleh kromatografi satah dengan pengunaan plat kromatografi lapisan
nipis (TLC) dan kromatografi lapisan nipis berprestasi tinggi (HPTLC). Fasa
pegun normal, terbalik dan silika terikat siano diuji dengan lima fasa bergerak
binari (asetonitril-air, asetonitril-DMSO, asetonitril-metanol,
petroleum-aseton eter, aseton-air) di mana kepekatan pengubahsuai organik
diubah daripada 0 hingga 100% (v/v).
Kajian ini melaporkan keadaan optimum bagi permisahan hormon steriod. Analisis
komponen prinsipal (PCA) berasaskan kiraan kuantitatif molekul yang membezakan
bahan larut dilakukan untuk menentukan persamaan dan perbezaan di antara
sebatian yang diuji. Keupayaan pemisahan fasa bergerak dan pegun dibandingkan
berdasarkan kepada faktor pemisahan α. Data tambatan kromatografi dan
kebarangkalian mekanisma tambatan juga turut dibincang.
Kata kunci: Kromatografi
lapisan nipis, faktor pemisahan, fasa binari, steroid
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