| Niuhuang Shang Qing tablet (NHSQ) which has been listed as Nation Essential Drug and incorporated in the 2010 edition of Chinese Pharmacopoeia is a widely used Traditional Chinese Medicine (TCMs) preparation for the treatment of clearing heat,relieving pain and dispelling wind.[2] It is composed of nineteen kinds of herbs,including Bov is Calculus Artifactus, Mentha haplocalyx Briq., Chrysanthemum morifolium, Schizonepetae Spica, Angelicae Dahuricae Radix, Ligusticum Chuanxiong Hort., Gardenia jasminoides Ellis., Coptis chinensis Franch.,Phellodendri chinensis Cortex., Scutellaria baicalensis Georgi, Rhei Radix et Rhizoma, Forsythia suspensa Vahl., Paeoniae Rubra Radix, Angelicae Sinensis Radix,Rehmannia glutinosa Libosch., Platycodon grandiflorum, Glycyrrhizae Radix et Rhizoma, Gypsum Fibrosum and Borneolum Syntheticum. It is a kind of Over-The-Counter (OTC) drug which is being produced on a large scale by many pharmaceutical companies in China. Up to now, however, there were no any reports about the pharmacokinetic studies of NHSQ. What the pharmacokinetic properties of rhein in NHSQ would be? Whether the different ingredients of NHSQ affect the metabolism and absorption of the major primsry in rats or not? Research should not stop there, the information of comparative study concerning the pharmacokinetic characteristics of rhein in pure chemical, single herb and different Chinese herbal formula as experimental drugs in vivo remain undiscovered. Therefore, the final goal of this project is to quantify the concentration of rhein in rat plasma and compare its pharmacokinetic parameters by oral administration of rhein, rhubarb extract and NHSQ, respectively.In this study, a new, simple, rapid, sensitive, convenient and economic high performance liquid chromatography with diode array detection (HPLC) method was established to determinate rhein in rats plasma after oral administration of NHSQ,rhubarb (Rheum palmatum L) extract and rhein reference substance, respectively with a linear range of 0.05-5.0 ?g/mL for rhein (r2=0.9991). 1, 8 - Dihydroxyanth-raquinone was used as an internal standard. The plasma samples were extracted byethyl acetate after acidification. Chromatographic separation was performed on an Inertsil(?)ODS-3 column (150 mm × 4.6 mm, 5 um) with a mobile phase consisting of methanol/ 0.2% phosphoric acid (80/20; v/v) at a flow rate of 1.0 mL/min. The method was validated and successfully applied to compare the pharmacokinetic characteristics of rhein in NHSQ with HLSQ, the rhubarb extract and rhein reference substance in rats after orally administration. This is the first investigation of pharmacokinetic data for rhein in NHSQ. The result of comparative pharmacokinetics of rhein instructed that the main parameters of the herbal formule (NHSQ) were significantly (p<0.05) different than that of the single herb (rhubarb extract) and the pure compound (rhein). The Cmax of the herbal formule (NHSQA: 2.462 mg/L; NHSQ B: 1.903 mg/L; NHSQc: 2.922 mg/L) were significantly (p<0.05) higher than that of the single herb (rhubarb extract: 1.116 mg/L) and the pure compound (rhein: 0.662 mg/L ). The Tmax of the herbal formule (NHSQA: 0.236 h; NHSQB: 0.333 h;NHSQc: 0.264 h) were significantly (p<0.05) less than that of the single herb(rhubarb extract: 0.542 h) and the pure compound (rhein: 0.958 h). Besides, the AUC of the herbal formule (NHSQA: 7.105 mg/L*h; NHSQB : 8.616 mg/L*h; NHSQC:8.385 mg/L*h) were significantly (p<0.05) higher than that of the single herb(rhubarb extract: 4.518 mg/L*h) and the pure compound (rhein: 2.395 mg/L*h). This conclusion revealed that the other herbal ingredients of NHSQ and HLSQ significantly enhance the absorption of rhein in rat plasma and different prescription makes unconspicuously different pharmacokinetic behavior of rhein in vivo, which makes NHSQ more efficient than the single herb and the pure compound in clinical applications of rhein.Physiological disposition of drug is significant work in drug research and development. The identification of bio active components and metabolites are conducive to interpret the action mechanism. In this study, an ultra high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) method was applied to identify the chemical constituents in NHSQ extracts and its absorbed components in rat plasma, urine and various tissues (brain,heart, liver and kidney) after oral administration of NHSQ. As a result, a total of 211 components of NHSQ extracts were identified in vitro, 135 prototype components and 154 metabolites of NHSQ in rat multiple biosamples were detected and identified by comparing with the NHSQ extracts and blank biosamples. Among the metabolites,five (M50, M79, M115, M128 and M154) of which were unreported previously. In addition, the metabolic pathways are proposed, including methylation, sulfation,oxidation, reduction, glucoside and glucuronidation. Among of them, glucuronide conjugates was the main metabolic pathway of the absorbed components in multi-biosamples. The results proved that the experimental strategy established is simple and reliable, which could provide a better comprehension of the ADME of TCM in vivo. |
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