Method development for Lawsone estimation in Trichup herbal hair powder by high-performance thin layer chromatography

Abstract

A simple, specific, accurate, precise and robust high-performance thin-layer chromatographic method has been developed and validated for estimation of Lawsone in Trichup herbal hair powder (coded as a THHP), polyherbal formulation. The chromatographic development was carried out on aluminum plates pre-coated with silica gel 60F254 and good resolution was achieved with Toluene: Ethyl acetate: Glacial acetic acid (8:1:1 v/v/v) as mobile phase. Lawsone detection was carried out densitometrically at 277 nm and obtained retardation factor value was 0.46 ± 0.02. The method was validated with respect to specificity, linearity, accuracy, precision and robustness. The calibration curve was achieved to be linear over a range of 5-60 μg/ml and regression coefficient was obtained 0.998. Accuracy of chromatographic method was evaluated by standard addition method; recovery was obtained 99.25 ± 0.61%. The peak purity of Lawsone was achieved 0.999 r. Relative standard deviation for intraday and inter-day precision was 0.37-0.56% and 0.42-0.55%, respectively. The limit of detection and limit of quantification of the Lawsone were found to be 1.08 μg/m land 3.28 μg/ml, respectively. This result shows that the method was well validated. In the present study, the Lawsone content was found 0.322 ± 0.014% in THHP. This study reveals that the proposed high performance thin layer chromatography method is accurate, fast and cost- effective for routine estimation of Lawsone in polyherbal formulation.

Keywords: Henna; Lawsone; Trichup herbal hair powder; high-performance thin-layer chromatography; validation.

Figure 1

Chemical structure of Lawsone

Figure 2

Chromatogram of 50 μg/ml Lawsone Standard

Figure 3

Chromatogram of Trichup herbal hair powder sample

Figure 4

3D graph for specificity. 1 - Lawsone std. 50 μg/ml, 2 - Henna, 3 - Henna, 4 - THHP, 5 - THHP, 6 - Placebo, 7 - Placebo, 8 - Placebo, 9 - Placebo

Figure 5

Overlay spectrum of standard Lawsone and sample

Figure 6

Overlay spectrum of standard Lawsone and placebo

Figure 7

Peak purity of Lawsone standard and Trichup herbal hair powder

Figure 8

3D graph for linearity. 1-5 μg/ml, 2-10 μg/ml, 3-20 μg/ ml, 4-30 μg/ml, 5-40 μg/ml, 6-50 μg/ml, 7-60 μg/ml

Figure 9

Calibration curve for Lawsone