Metabolic profile of salidroside in rats using high-performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry.

A high-performance liquid chromatography coupled to Fourier transform ion cyclotron resonance mass spectrometry (HPLC-FT-ICR MS) method was developed to study the in vivo metabolism of salidroside for the first time. Plasma, urine, bile, and feces samples were collected from male rats after a single intragastric gavage of salidroside at a dose of 50 mg/kg. Besides the parent drug, a total of seven metabolites (three phase I and four phase II metabolites) were detected and tentatively identified by comparing their mass spectrometry profiles with those of salidroside. Results indicated that metabolic pathways of salidroside in male rats included hydroxylation, dehydrogenation, glucuronidation, and sulfate conjugation. Among them, glucuronidation and sulfate conjugation were the major metabolic reactions. And most important, the detection of the sulfation metabolite of p-tyrosol provides a clue for whether the deglycosylation of salidroside occurs in vivo after intragastric gavage. In summary, results obtained in this study may contribute to the better understanding of the safety and mechanism of action of salidroside.

Effect of liquid-to-solid lipid ratio on characterizations of flurbiprofen-loaded solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) for transdermal administration.

The aim of this study is to evaluate the effect of liquid-to-solid lipid ratio on properties of flurbiprofen-loaded solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs), and to clarify the superiority of NLCs over SLNs for transdermal administration. Particle size, zeta potential, drug encapsulation efficiency, in vitro occlusion factor, differential scanning calorimetry, X-ray diffractometry, in vitro percutaneous permeation profile, and stability of SLNs and NLCs were compared. Particle size, zeta potential, drug encapsulation efficiency, in vitro occlusion factor, and in vitro percutaneous permeation amount of the developed NLCs were all <200 nm, < -20 mV, >78%, >35, and >240 µg/cm(2), respectively, however, for SLNs were 280 nm, -29.11 mV, 63.2%, 32.54, and 225.9 µg/cm(2), respectively. After 3 months storage at 4 °C and 25 °C, almost no significant differences between the evaluated parameters of NLCs were observed. However, for SLNs, particle size was increased to higher than 300 nm (4 °C and 25 °C), drug encapsulation efficiency was decreased to 51.2 (25 °C), in vitro occlusion factor was also decreased to lower than 25 (4 °C and 25 °C), and the cumulative amount was decreased to 148.9 µg/cm(2) (25 °C) and 184.4 µg/cm(2) (4 °C), respectively. And DSC and XRD studies indicated that not only the crystalline peaks of the encapsulated flurbiprofen disappeared but also obvious difference between samples and bulk Compritol® ATO 888 was seen. It could be concluded that liquid-to-solid lipid ratio has significant impact on the properties of SLNs and NLCs, and NLCs showed better stability than SLNs. Therefore, NLCs might be a better option than SLNs for transdermal administration.

Characterization of chemical constituents in Zhi-Zi-Da-Huang decoction by ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

A sensitive and reliable ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry method was established to separate and identify the chemical constituents of Zhi-Zi-Da-Huang decoction, a classic traditional Chinese medicine formula. The chromatographic separation was achieved on a Shim-pack XR-ODS C18 column (75 × 3.0 mm, 2.2 µm) using a gradient elution program. The detection was performed on a Waters Xevo G2 Q-TOF mass spectrometer equipped with electrospray ionization source in both positive and negative modes. With the optimized conditions, a total of 82 compounds were identified or tentatively characterized. Of the 82 compounds, 21 compounds were identified by comparing the retention time and MS data with reference standards, the rest were characterized by analyzing MS data and retrieving the reference literature. In addition, 31 compounds were identified from Gardenia jasminoides Ellis, ten compounds were identified from Rheum palmatum L., 33 compounds were identified from Citrus aurantium L., and eight compounds were identified from Sojae Semen Praeparatum. Results indicated that iridoids, anthraquinones, flavonoids, isoflavonoids, coumarins, glycosides of crocetin, monoterpenoids, and organic acids were major constituents in Zhi-Zi-Da-Huang decoction. It is concluded that the developed ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry method with high sensitivity and resolution is suitable for identifying and characterizing the chemical constituents of Zhi-Zi-Da-Huang decoction, and the analysis provides a helpful chemical basis for further research on Zhi-Zi-Da-Huang decoction.

Characterization of chemical constituents in Rhodiola Crenulate by high-performance liquid chromatography coupled with Fourier-transform ion cyclotron resonance mass spectrometer (HPLC-FT-ICR MS).

In this work, an approach using high-performance liquid chromatography coupled with diode-array detection and Fourier-transform ion cyclotron resonance mass spectrometer (HPLC-FT-ICR MS) for the identification and profiling of chemical constituents in Rhodiola crenulata was developed for the first time. The chromatographic separation was achieved on an Inertsil ODS-3 column (150 mm × 4.6 mm,3 µm) using a gradient elution program, and the detection was performed on a Bruker Solarix 7.0 T mass spectrometer equipped with electrospray ionization source in both positive and negative modes. Under the optimized conditions, a total of 48 chemical compounds, including 26 alcohols and their glycosides, 12 flavonoids and their glycosides, 5 flavanols and gallic acid derivatives, 4 organic acids and 1 cyanogenic glycoside were identified or tentatively characterized. The results indicated that the developed HPLC-FT-ICR MS method with ultra-high sensitivity and resolution is suitable for identifying and characterizing the chemical constituents in R. crenulata. And it provides a helpful chemical basis for further research on R. crenulata. Copyright © 2016 John Wiley & Sons, Ltd.

Evaluation of the protective effect of Zhi-Zi-da-Huang decoction on acute liver injury with cholestasis induced by α-naphthylisothiocyanate in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Zhi-Zi-Da-Huang decoction (ZZDHD), a classic traditional Chinese medicine (TCM) formula composed of four herbal medicines, has been widely used to treat various hepatobiliary disorders for a long time in China. However, the pharmacological effect of ZZDHD on liver injury with cholestasis is unrevealed. AIM OF THE STUDY: To investigate the hepatoprotective effect of ZZDHD against α-naphthylisothiocyanate (ANIT)-induced liver injury with cholestasis in rats. MATERIALS AND METHODS: The rats were intragastrically (i.g.) given ZZDHD at doses of 1, 2 and 4 g/kg (crude drug/body weight) once a day for seven days and treated with ANIT (75 mg/kg via i.g.) to cause liver injury at 12h after the fifth administration. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyltranspeptidase (γ-GTP), total bilirubin (TBIL), direct bilirubin (DBIL) and total bile acid (TBA), as well as bile flow were measured at 48 h after ANIT treatment to evaluate the protective effect of ZZDHD. Moreover, the possible protective mechanisms were elucidated by assays of liver enzyme activities and component contents including malondialdehyde (MDA), myeloperoxidase (MPO), lipid peroxide (LPO), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). The biochemical observations were supplemented by histopathological examination. Ultra fast liquid chromatography-mass spectrometry (UFLC-MS) was used for the phytochemical analysis of ZZDHD. RESULTS: The high dose (4 g/kg) and middle dose (2g/kg) of ZZDHD exhibited significant and dose-dependent protective effect on ANIT-induced liver injury with cholestasis by reversing the changes in bile flow, the serum and hepatic enzymes, and histopathology of the liver tissue. Meanwhile, it was found that the low dose (1g/kg) of ZZDHD did not improve the biochemical indexes except serum TBIL, DBIL and TBA, which showed little protective effect. Phytochemical analysis revealed the presence of sixteen compounds in ZZDHD. CONCLUSIONS: This study indicates that ZZDHD exerted a hepatoprotective effect on ANIT-induced liver injury with cholestasis in rats, and the mechanism of this activity is possibly related to its antioxidant properties.

Identification of the absorbed components and metabolites of Zhi-Zi-Da-Huang decoction in rat plasma by ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry.

Zhi-Zi-Da-Huang decoction (ZZDHD), consisting of Gardenia jasminoides Ellis, Rheum palmatum L., Citrus aurantium L. and Sojae Semen Praeparatum, is a widely used traditional Chinese medicine preparation for the treatment of acute or chronic hepatic diseases. In the present study, a sensitive and selective ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) method was developed to separate and identify the absorbed components and metabolites in rat plasma after oral administration of ZZDHD. The plasma samples were pretreated by protein precipitation and separated on a Shim-pack XR-ODS C18 column (75 mm × 3.0 mm, 2.2 µm) using a gradient elution program. Mass spectrometric detection was performed on an Agilent 6520 Q-TOF mass spectrometer equipped with electrospray ionization (ESI) source in positive and negative ion modes. By comparing the retention time, high resolution mass data of blank plasma and dosed plasma, a total of 43 constituents, including 21 prototype compounds and 22 metabolites were identified or tentatively characterized. Results indicated that glucuronidation and sulfation were the main metabolic pathways of iridoid glycosides and anthraquinones, glucuronidation was the main metabolic pathways of flavanone-related compounds. It is concluded the developed UHPLC-Q-TOF-MS method with high sensitivity and resolution is suitable for identifying and characterizing the absorbed components and metabolites of ZZDHD, and the results will provide essential data for further studying the relationship between the chemical components and pharmacological activity of ZZDHD.