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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (1): 109-113.doi: 10.3736/jcim20120116

• Original Experimental Research • Previous Articles     Next Articles

Isolation, characterization and thin-layer chromatography method development of clerosterol palmityl ester: a chemical marker for standardization of leaves of Clerodendrum phlomidis

Kumaradoss Mohan Maruga Raja Muthu1(),Hari Mishra Shri2   

  1. 1. Department of Pharmacognosy and Phytochemistry, Karnataka Kaveri Educational and Cultural Society College of Pharmacy, Bangalore 560103, Karnataka, India
    2. Department of Pharmacy, The Maharaja Sayajirao University of Baroda, Vadodara 390002, Gujarat, India
  • Received:2011-09-28 Accepted:2011-11-09 Online:2012-01-20 Published:2018-09-21
  • Contact: Kumaradoss Mohan Maruga Raja Muthu E-mail:mohanmarugaraja@gmail.com

OBJECTIVE: Clerosterol palmityl ester (CPE) is a unique clerosterol derivative isolated and characterized from the leaves of Clerodendrum phlomidis. Considering the uniqueness of this compound, the present study was planned to use CPE as a specific chemical marker and develop a new validated thin-layer chromatography (TLC) method for standardisation of C.phlomidis.METHODS: Separation and quantification of CPE were achieved by TLC using a mobile phase of petroleum ether (60 to 80 ℃) and ethyl acetate (95∶5, volume ratio) (Rf 0.64) on precoated silica gel 60F254 aluminium plates. Densitometric determination was carried out after derivatization with anisaldehyde sulphuric acid reagent in absorption mode at 527 nm.RESULTS: The calibration curve was linear in the concentration range of 100 to 500 ng/spot. The method was validated for precision, repeatability and accuracy. The proposed method was found to be simple, specific, precise, accurate, rapid and cost-effective.CONCLUSION: This TLC procedure may be used effectively for quantitative determination of CPE, identification of the plant and standardization of this plant or its derived products.

Key words: chromatography, thin-layer, Clerodendrum, plant extracts

Figure 1

Structure of clerosterol palmityl ester"

Figure 2

Densitogram of the ST and CPEST: sample track; CPE: clerosterol palmityl ester."

Figure 3

Spectral comparison for the peaks of ICPE and CPE in C. phlomidis leaf extractICPE: isolated clerosterol palmityl ester; CPE: clerosterol palmityl ester."

"

Parameter Result
Rf 0.64
Dynamic range (ng/spot) 100 to 500
Equation Y=480.006+10.106X
Slope 10.106
Intercept 480.006
Limit of detection 15.49 ng
Limit of quantification 46.95 ng
Linearity (correlation coefficient) 0.999

"

Method precision (%RSD)
Concentration
(ng/spot)
Instrumental precision
(%RSD)
Intra-day Inter-day
100 0.58 0.79 1.31
500 0.17 0.22 0.37

"

Amount of CPE
in the sample (μg)
Amount of CPE
added (μg)
Amount of
CPE found (μg)
Recovery
(%)
95.79 76.62 165.24 95.84
95.79 95.80 184.71 96.41
95.79 114.90 209.63 99.50
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