Simultaneous quantification of some pharmaceutical Catharanthus roseus leaf and root terpenoid indole alkaloids and their precursors in single runs by reversed-phase liquid chromatography.
A new, rapid, sensitive, and reproducible reversed-phase liquid chromatographic (LC) method with photodiode array detection is described. It allows, in a single run of 30 min, simultaneous separation of 6 pharmaceutically and biologically important Catharanthus roseus leaf and root terpenoid indole alkaloids (TIAs) and 3 of their precursors: TIA precursors tryptophan, tryptamine, and loganine; and TIAs serpentine, catharanthine, ajmalicine, vincristine, vinblastine, and vindoline. The method involves the use of a Phenomenex Luna 5 microm, C18 column (250 mm x 4.6 mm id) and a linear binary gradient mobile phase profile. Detection is performed at 220 and 254 nm, which provided good absorptivity for all of the roseus compounds listed above and gave a minimum detection limit of 0.02 microg/mL. The extraction efficiency, peak purity, and homogeneity parameters of the profiles could be validated using a photodiode array detector. The method was successfully used to quantify major components of leaf and root extracts of C. roseus accessions. The new method thus provides a reliable tool for rapid screening of C. roseus samples in large numbers, which is needed in breeding/genetic engineering and genetic mapping experiments and for monitoring the reaction products, in the in vitro/in vivo conversions of precursors into products, and vice versa.
Digvijay
Singh
S
National Center for Plant Genome Research, Jawaharlal Nehru University Campus, New Delhi-110 067, India.
Shashi
Pandey-Rai
PR
Suchi
Srivastava
S
Kumar
Rai Sanjay
RS
Raghavendra
Mishra
M
Sushil
Kumar
K
D000470:Alkaloids / Q000032:analysis*; D030008:Catharanthus / Q000737:chemistry*; D002853:Chromatography, Liquid; D007202:Indicators and Reagents; D007211:Indoles / Q000032:analysis*; D010936:Plant Extracts / Q000032:analysis; D018515:Plant Leaves / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D012996:Solutions; D013729:Terpenes / Q000032:analysis*
D000470:Alkaloids; D007202:Indicators and Reagents; D007211:Indoles; D010936:Plant Extracts; D012996:Solutions; D013729:Terpenes