RESUMEN
Zhi-zi-chi decoction (ZZCD) is a classical formula widely used in Chinese clinical application. In the present study, a novel and efficient strategy has been developed for screening and identification of multiple constituents and their metabolites of ZZCD using ultra-high-performance liquid chromatography combined with triple time-of-flight mass spectrometry. The novel approach of an online data acquisition method dependent on multiple mass defect filter and dynamic background subtraction is combined with multiple data processing techniques. First, a total of 109 potential bioactive compounds were detected in ZZCD. Based on the same instrumental conditions, 100 compounds were found in rat biofluids after oral administration of ZZCD, including 61 original compounds of ZZCD as well as 39 metabolites. Conjugations with sulfate, glucuronate and amino acids were found as the predominant metabolic reaction of ZZCD. As more xenobiotics were detected in urine than those in bile were, it demonstrated that multiple components of ZZCD have undergone comprehensive renal excretion. This study reported the urinary and biliary excretion in rats after oral administration of ZZCD for the first time. The present study expands our knowledge about the constituents and metabolism of ZZCD, which could be very useful for further pharmacological and clinical studies of ZZCD.
Asunto(s)
Bilis/química , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/metabolismo , Espectrometría de Masas en Tándem/métodos , Animales , Medicamentos Herbarios Chinos/química , Glicósidos Iridoides/análisis , Glicósidos Iridoides/química , Glicósidos Iridoides/metabolismo , Glicósidos Iridoides/orina , Ratas , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
There is a growing concern for the sensitive quantification of multiple components using advanced data acquisition method in herbal medicines (HMs). An improved and rugged UPLC-MS/MS method has been developed and validated for sensitive and rapid determination of multiply analytes from Tong-Xie-Yao-Fang (TXYF) decoction in three biological matrices (plasma/brain tissue/urine) using geniposide and formononetin as internal standards. After solid-phase extraction, chromatographic separation was performed on a C18 column using gradient elution. Quantifier and qualifier transitions were monitored using novel Triggered Dynamic multiple reaction monitoring (TdMRM) in the positive ionization mode. A significant peak symmetry and sensitivity improvement in the TdMRM mode was achieved as compared to conventional MRM. The reproducibility (RSD%) was ≤7.9% by applying TdMRM transition while the values were 6.8-20.6% for MRM. Excellent linear calibration curves were obtained under TdMRM transitions over the tested concentration ranges. Intra- and inter-day precisions (RSD%) were ≤14.2% and accuracies (RE%) ranged from -9.6% to 10.6%. The validation data of specificity, carryover, recovery, matrix effect and stability were within the required limits. The method was effectively applied to simultaneously detect and quantify 1 lactone, 2 monoterpene glucosides, 1 alkaloid, 5 flavonoids and 2 chromones in plasma, brain tissue and urine after oral administration of TXYF decoction. In conclusion, this new and reliable method is beneficial for quantification and confirmation assays of multiply components in complex biological samples.