Biosynthesis of antitumoral and bactericidal sanguinarine

Abstract

A simple, rapid, and reliable TLC method for the separation and determination of sanguinarine has been established. This intensively studied biologically active alkaloid has a wide range of potentially useful medicinal properties, such as antimicrobial, antiinflammatory, and antitumoral activities. Sanguinarine has also been incorporated into expectorant mixtures and has a strong bactericidal effect upon gram-positive bacteria, particularly Bacillus anthracis and staphylococci. These medicinal properties are due to the interaction of sanguinarine with DNA. A fibre-optic-based fluorescence instrument for in situ scanning was used for quantitative measurements. The sanguinarine was determined over the range 5-40 ng and a detection limit of 1.60 ng. The method was applied to the quantification of sanguinarine in tissue culture extracts of Chelidonium majus L.

Figure 1

Excitation (1,2) and emission (1′,2′) spectra of sanguinarine. (1,1′) 3 mg/L of sanguinarine in aqueous medium, slits 5 nm, λ_exc = 283 nm, and λ_em = 415 nm. (2,2′) 1 mg/L of sanguinarine in ethanol medium, slits 2.5 nm, λ_exc = 283 nm, and λ_em = 412 nm. In situ excitation (3,4) and emission (3′,4′) spectra of sanguinarine, obtained with the fibre-optic sensor after elution on a silica gel plate. (3,3′) 20 ng and (4,4′) 100 ng of sanguinarine from ethanol and aqueous solutions, respectively. Slits 10 nm, λ_exc = 326 nm, and λ_em = 545 nm.

Figure 2

Contour map 2(a) and three-dimensional chromatogram 2(b) scanned at λ_exc = 326 nm and λ_em = 545 nm (slits 10 nm) for seven samples of sanguinarine ranged from 5 to 40 ng, after they were eluted with hexane : ethyl acetate : ammonia (25%) (6 : 4 : 0.1) (v : v : v) on a silica gel plate.

Figure 3

Shewhart chart for control of the chromatographic analysis of sanguinarine. UCL and LCL: upper and lower control limits. UWL and LWL: upper and lower warning limits. Mean = 19.72, S = 1.66.

Figure 4

Molecular structure of sanguinarine. One of the best targets for testing interaction of a drug with its receptor is DNA. Antitumor antibiotics are known to act at the DNA level. Due to DNA's helical and relatively stable conformation in solution, DNA-drug complexes can conveniently be studied by high-resolution NMR-spectroscopy.