This paper describes the development of an HPLC-UV-MS method for quantitative determination of andrographolide and dehydroandrographolide in Andrographis Herba and establishment of its chromatographic fingerprint. The method was validated for linearity, limit of detection and quantification, inter- and intra-day precisions, repeatability, stability and recovery. All the validation results of quantitative determination and fingerprinting methods were satisfactory. The developed method was then applied to assay the contents of andrographolide and dehydroandrographolide and to acquire the fingerprints of all the collected Andrographis Herba samples. Furthermore, similarity analysis and principal component analysis were used to reveal the similarities and differences between the samples on the basis of the characteristic peaks. More importantly, the DPPH free radical-scavenging and ferric reducing capacities of the Andrographis Herba samples were assayed. By bivariate correlation analysis, we found that six compounds are positively correlated to DPPH free radical scavenging and ferric reducing capacities, and four compounds are negatively correlated to DPPH free radical scavenging and ferric reducing capacities.

In the present study, we investigated the tissue distribution and urinary excretion of gallic acid (GA) and protocatechuic acid (PCA) after rat oral administration of aqueous extract of Polygonum capitatum (P. capitatum, named Herba Polygoni Capitati in China). An UHPLC-MS/MS analytical method was developed and adopted for quantification of GA and PCA in different tissue homogenate and urine samples. Interestingly, we found that GA and PCA showed a relatively targeted distribution in kidney tissue after dosing 60 mg/kg P. capitatum extract (equivalent to 12 mg/kg of GA and 0.9 mg/kg of PCA). The concentrations of GA and PCA in the kidney tissue reached 1218.62 ng/g and 43.98 ng/g, respectively, at one hour after oral administration. The results helped explain the empirical use of P. capitatum for kidney diseases in folk medicine. Further studies on urinary excretion of P. capitatum extract indicated that GA and PCA followed a concentrated elimination over a 4-h period. The predominant metabolites were putatively identified to be 4-methylgallic acid (4-OMeGA) and 4-methylprotocatechuic acid (4-OMePCA) by analyzing their precursor ions and characteristic fragment ions using tandem mass spectrometry. However, the amount of unchanged GA and PCA that survived the metabolism were about 14.60% and 15.72% of the total intake, respectively, which is reported for the first time in this study.

Citri Exocarpium Rubrum (CER), which is known as Juhong in Chinese, is the dried exocarp of Citrus reticulata Blanco and its cultivars (Fam. Rutaceae) and is currently used in Chinese medicine to protect the stomach and eliminate dampness and phlegm. The main aim of this study was to develop a high-performance liquid chromatography ultraviolet mass spectrometry (HPLC-UV-MS) method for determining the chemical compositions and fingerprint of CER. We also evaluated the antioxidant properties of CER based on its 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, ferric ion reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC) assays. Ten CER samples were collected from Hong Kong and mainland China. Each CER sample was extracted using an ultrasonic extraction method. Chromatographic separation was achieved using a conventional Dikma Inspire C18 column with photo diode array detection (190–400 nm). Hesperidin, nobiletin and tangeretin were quantified based on the UV signal observed at 330 nm. The column was eluted with a mobile phase consisting of water and acetonitrile (15–55%) over 55 min. Fingerprints combined with similarity and principal component analyses were used to classify the herbs. The DPPH free radical scavenging activity, FRAP and ABTS properties of the different CER samples were assayed. Bivariate correlation analysis was performed to investigate the correlation between the characteristic peaks and their antioxidant capacities. Limit of detection (LOD), limit of quantification (LOQ), linearity, inter-day precision, intra-day precision, repeatability, stability and recovery of the developed method were validated, and the method was subsequently used to determine the contents of hesperidin, nobiletin and tangeretin, and to acquire the fingerprints of the CER samples. Seventeen characteristic peaks were found in the fingerprints, and eleven of them were identified. Bivariate correlation analysis revealed correlations between the characteristic peaks and the antioxidant activities of the samples. An HPLC-UV-MS method was developed and validated after a detailed investigation on extraction of chemical compounds from CER using different solvents and extraction times. None of the peaks was correlated with the DPPH free radical scavenging activity or ferric reducing capacity. Most of the peaks were correlated well with the ABTS radical scavenging capacity.

Background: Polygoni Multiflori Caulis, the dried caulis of Polygonum multiflorum Thunb., is one of the commonly used traditional Chinese medicines having antioxidant, anti-obesity, anti-inflammatory and antibacterial effects. Polygoni Multiflori Caulis used clinically or circulated on market have great differences in their diameters. However, to the best of our knowledge, no study has been reported on the qualities of Polygoni Multiflori Caulis with different diameters. Results: Systematic HPLC/UV/MS chromatographic fingerprinting and quantitative analytical methods combined with principal component analysis were developed and applied to analyze different Polygoni Multiflori Caulis samples. The contents of 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside, the chemical marker for quality control on Polygoni Multiflori Caulis specified in Chinese Pharmacopoeia (2010 edition), were found to have surprising relevance with the samples’ diameters for the first time. Conclusion: The finding provides a scientific basis for collecting Polygoni Multiflori Caulis in the best time. Moreover, the diameter can be used as the criterion for quality control on Polygoni Multiflori Caulis as a preliminary step in the future. In addition, scores plot obtained from principal component analysis shows the obvious differences between unqualified Polygoni Multiflori Caulis samples and qualified ones visually, which can be used to single out the unqualified ones with qualified ones efficiently and immediately.

The present paper describes the development of a high performance liquid chromatography-ultraviolet (HPLC-UV) detection method for quantitative determination of peimine and peiminine in Fritillariae Thunbergii Bulbus (FTB). Separation was achieved using a conventional XBridge™ Shield RP 18 column (250 mm × 4.6 mm, internal diameter 3.5 μm) with photodiode array detection at 190-400 nm for UV spectra and 220 nm for quantification. The mobile phase consisted of (A) 0.03% diethylamine aqueous solution and (B) acetonitrile eluted by an isocratic procedure at 45:55 (A:B) over 25 minutes. The method was validated for linearity, limits of detection (LOD) and quantification (LOQ), inter- and intra-day precisions, repeatability, stability, and recovery. All the validation results were satisfactory. The developed method was then applied to assay the contents of the two chemical markers in all the FTB samples collected. Based on the contents of the two analytes, hierarchical clustering analysis (HCA) was performed to reveal the similarities and differences of the samples.