title: Determination of total retronecine esters-type hepatotoxic pyrrolizidine alkaloids in plant materials by pre-column derivatization high-performance liquid chromatography.
abstract: A pre-column derivatization high-performance liquid chromatography method with diode array detection was developed and validated to determine the total retronecine esters-type hepatotoxic pyrrolizidine alkaloids (RET-HPAs) in herbs. The RET-HPAs reacted with o-chloranil in methanolic solution heated for 3 h, and an oxidative derivative was produced that could be detected at a maximal absorption of 223 nm. The analysis was performed using a C18 column with an isocratic elution of methanol and aqueous 0.01% triethylamine (adjusted to pH 4 with formic acid), and the detection was carried out with DAD at 223 nm. The validation of the method included linearity, sensitivity, recovery and stability. It showed a good linear regression (r(2) > 0.9900) in the range of 2.5-250 microM with a limit of detection (S/N = 3) of 0.5 microM. The method provided desirable repeatability with overall intra- and inter-day variations of less than 4.6%. The obtained recoveries for both of the extraction and derivatization process were between 94.6 and 100.7% (n = 3).
authors.affiliation: Key Laboratory of Standardization of Chinese Medicines of Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, People's Republic of China.
authors.forename: Ai-Zhen
authors.initials: AZ
authors.lastname: Xiong
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Fang
authors.initials: F
authors.lastname: Zhang
authors.forename: Xue-Jing
authors.initials: XJ
authors.lastname: Yang
authors.forename: Chang-Hong
authors.initials: CH
authors.lastname: Wang
authors.forename: Zheng-Tao
authors.initials: ZT
authors.lastname: Wang
mesh_terms: D002703:Chloranil / Q000031:analogs & derivatives / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D006358:Hot Temperature; D006801:Humans; D000432:Methanol / Q000737:chemistry; D010084:Oxidation-Reduction; D010936:Plant Extracts / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D010946:Plants, Medicinal / Q000737:chemistry*; D011763:Pyrrolizidine Alkaloids / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014118:Toxins, Biological / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D010936:Plant Extracts; D011763:Pyrrolizidine Alkaloids; D014118:Toxins, Biological; D002703:Chloranil; C014795:retronecine; C039499:2-chloranil; D000432:Methanol