Simultaneous Determination of Paclitaxel and 10-Deacetyl Baccatin in Taxus Baccata Bark by HPTLC

Background and Aim:The aim of this study was to develop an HPTLC method for the simultaneous estimation of paclitaxel (PAC) and 10-deacetyl baccatin (DAB) in Taxus baccata bark.

Methods: Quantitative estimation of PAC and DAB in the methanolic extract of the bark of T. baccata and its dichloromethane fraction was aimed in the study. Pre-coated silica gel 60 F₂₅₄ plates were used as 254 stationary phase and chloroform: acetonitrile (7:3 v/v) as the mobile phase. Standards were dissolved in analytical grade methanol separately to prepare stock solutions (1mg/ml, each). Calibration plots ranging from 1μl to 7μl were prepared. Densitometric detection and quantification of PAC and DAB was carried out at 254 nm.

Result: The HPTLC analysis showed the presence of PAC and DAB as compact spots at Rf of 0.48 ± 0.01 and 0.22 ± 0.01 respectively. The calibration plots were found to be linear for PAC and DAB with a regression coefficient of 0.98. The results indicated sufficient peak linearity and purity. The method was found to be precise, robust, specific and accurate. The results are compiled in Table 1 and the HPTLC plates are shown in Figure 1.

Conclusion: An efficient HPTLC method for the simultaneous estimation of PAC and DAB in T. baccata bark was developed. The present HPTLC method can be used for their simultaneous quantification in plant extracts and fermented products.

Fig. 1:

HPTLC fingerprints of Taxus baccata leaves visualized at 254 nm [Mobile phase Chloroform-Acetonitrile (7:3)]

Table 1:

Content of DAB and taxol as estimated by HPTLC

DAB	Volume (μl)	Conc. (μg)	RF	Taxol	Volume (μl)	Conc. (μg)	RF
Extract	6	0.761	0.23	Extract	5	0.253	0.49
DCM fraction	6	1.169	0.23	DCM fraction	5	0.430	0.47