ABSTRACT
Fifteen compounds were isolated from the toad skin by a combination of various chromatographic methods including macroporous resin, silica gel, ODS and semi-preparative HPLC. Their structures were identified as 4,5-dimethyl-1,3,4,5-tetrahydropyrrolo[4,3,2-de]quinolin-6-ol(1), serotonin(2), N-methyl serotonin(3), O-methyl bufotenine(4), 1,2,3,4-tetrahydro-6-hydroxy-β-carboline(5), O-methylserotonin(6), glycinebetaine(7), caffeine(8), bufotenine(9), shepherdine(10), tryptophan(11), (5-hydroxy-1H-indol-3-yl)acetic acid(12), 5-hydroxy tryptophol(13), 2-methyl-6-hydroxy-1,2,3,4-tetrahydro-β-carboline(14), bufothionine(15). Among them, compound 1 was a new compound,compound 5 was a new natural product. Compounds 4-8 and 10-14 were separated from toad skin for the first time.
ABSTRACT
AIM:To establish a simple and accurate HPLC method for quality evaluation and determination of 8 active constituents, i.e. Danshensu, Protocatechualdehyde,rosmarinic acid,Salvianolic acid B, Dihydrotanshinone, Cryptotanshinone, Tanshinone Ⅰand Tanshinone ⅡA in Radix Salviae Miltiorrhizae, at the same time, in order to study the correlations of the contents of the 8 constituents. METHODS: Detected at 286 nm, analytes were separated on a ODS C_ 18 column(150 mm?4.6 mm, 10 ?m) eluted gradually with methanol and 1% tetrafuran in 1% formic acid water solution(3∶97)on the increasement of methanol from 3% to 70% in the whole analysis. After determination, SPSS 13.0 software was used to calculate the correlation between the 8 constituents. RESULTS: Under the above conditions, HPLC method was constructed to quantify 8 constituents,and their correlation coefficients were calculated. CONCLUSION: The result shows that the contents of the 8 constituents in Salvia miltiorrhiza Bge. vary greatly from different origins; and there are significant correlations between the contents of hydrophilic constituents such as Salvianolic acid B,Danshensu,rosmarinic acid, so do hydrophobic constituents like Cryptotanshinone,Tanshinone Ⅰ,Tanshinone ⅡA,but there is no correlation between the hydrophilic constituents and hydrophobic constituents.