OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.

Objective To establish quality control of Xinnaoling granules .Methods Thin layer chromatography (TLC) methods were used to identify Rhizoma Corydalis ,Radix Angelicae dahuricae ,Cortex Magnoliae officinalis ,Radix Glycyrrhi-zae ,Rhizoma Ligusticum ,Ligusticum wallichii ,Schisandra Chinensis ,Dendranthema morifolium .The concentration of salvi-anolic acid B was determined by high performance liquid chromatography (HPLC) .The method employed a column of Agilent Eclipse XDB-C18 (4 .6 mm × 250 mm ,5 μm) with a mobile phase of 0 .5% formic acid (A)-acetonitrile (B) at a temperature 25℃ .The gradient elution program was as follow :0~5 min 24% B ,5~6 min 24% ~19% B ,6~25 min 19~20% B .The flow rate was 1 .0 ml/min ,and the injection volume was 10μl and the detection wavelength was 286 nm .Results The spots in TLC plates were clear and specific .As for salvianolic acid B ,the linear range was 20-1 280 μg/ml and the equation of linear regres-sion of salvianolic acid B was Y=13 .304 X -117 .50 (r=0 .999 9 ,n=7) .The average recovery rate was 96 .17% (RSD=1 . 10% ) .Conclusion The method was proved to be simple ,reliable ,reproducible ,and could be used in the quality control of Xinnaoling g ranules .

Human cytochrome P450 (CYP) 3A ,which is widely involved in the various drug metabolism ,is most abun-dant in liver and intestine .The activity of CYP3A enzyme may be induced or inhibited in the process of drug metabolisms ,and affect the metabolism of other CYP3A substrates and modulators vice versa .At present ,in vitro probe drugs and in vivo bio-markers are both available to evaluate the activity of CYP 3A enzyme .The former requires oral probe drugs ,the latter does not need for those drugs and just allows laboratory technicians to detect endogenous substrates ,such as 4β-hydroxycholesterol and 6β-hydroxycortisol .As reported ,studies on CYP3A help to explain the inter-individually variability in drug metabolism ,to in-dicate dose adjustments in combination regimens when drug interactions exist ,to predict drug efficacy and toxicity reaction for providing theoretical guidance for individualized medication ,and to reduce market risk of new drugs for the potential drug inter-actions .We summarized these two kinds of endogenous biomarkers and their clinical application in this review .

Objective To establish a quality control standard for Qingre Baidu granules .Methods Isatidis Radix ,Fruc-tus Forsythiae ,Herba Violae ,and Glycyrrhizae were identified by TLC ,and the concentration of chlorogenic acid was deter-minedbyHPLC.ThismethodwasemployedonanAgilentZORBAXSB-C18column(4.6mm×250mm,5μm)at30℃ witha mobile phase of acetonitrile (A) and 0 .2% formic acid (B) using the gradient elution program shown as follows :0-12 min , 11%-12% A run at the flow rate of 1 .0 ml/min .The injection volume was 20 μl and the detection wavelength was 327 nm . Results Characteristic spots could be detected by TLC and the specificity of the method was satisfactory .As for chlorogenic acid ,the equation of linear regression of chlorogenic acid was Y=60 .239 4X+9 .096 3 (r=0 .999 9) with the linear range of 6.19-396 .00 μg/ml .The average recovery was 99 .66% (RSD=2 .82% ) .Conclusion The established method is simple ,reli-able ,reproducible ,and can be used for the quantitative determination and quality control of Qingre Baidu granules .

Objective To confirm the hepatotoxicity of valproic acid (VPA ) and its metabolites (2-propyl-4-pentenoic acid ,3-hydroxy valproic acid ,5-hydroxy valproic acid) on human liver cells .Methods Cells were divided into control group and VPA-treated group .The control group was conventionally cultured while the VPA-treated group was treated with valproic acid and its metabolites . The rate of cell proliferation was assayed by CCK 8 protocol . The mRNA levels of CYP1A1 , CYP1A2 ,PCNA ,Bax and Bcl-2 were measured by real time PCR .The correlated protein levels were measured by Western Blotting .The activity of LDH ,AST and ALT were also detected .Results Compared to the control group ,with the increases of concentrations and reaction time of VPA and its metabolites ,the proliferation rate of L02-cell was reduced ,the mRNA and protein levels of CYP1A1 ,CYP1A2 ,and Bax was increased ,the mRNA and protein level of PCNA and Bcl-2 was decreased , AST ,ALT ,and LDH were also elevated in the treated group .Conclusion Valproic acid and its metabolites were positively re-lated to hepatotoxicity .

Objective To establish the quality standard of Santeng oral solution.Method Sargentodoxa Caulis and Spatholobi Caulis were identified by thin layer chromatography.Chlorogenic acid was assayed by high performance liquid chromatography.The chromatographic column is Agilent Zorbax SB C18 (4.6 mm×250 mm, 5 μm) with a stable temperature of 35 ℃.The mobile phase in isocratic elution consists of acetonitrile and 0.1% folic acid aqueous solution with a preliminary volume ratio of 9∶91.The flow rate is 1.0 ml/min with an injection volume of 20 μl.Results Thin layer chromatography showed distinct spots of Sargentodoxa Caulis and Spatholobi Caulis with a great specificity.A regression formula Y=60.14X-6.37(r>0.999 9) was obtained with a good linearity in concentration range of 2.70~202.50 μg/ml.Conclusion A simple, stable and repeatable method was established for the quality control of Santeng oral solution.

OBJECTIVE: To study the effects of Wuzhi capsule/schisantherin A (SchA) combined with cyclophosphamide on the pharmacokinetics of cyclophosphamide (CTX) in rats. METHODS: A total of 36 rats were randomly divided into CTX group (via tail vein, iv, CTX solution 300 mg/kg), CTX+WZC group (ig, Wuzhi capsule 300 mg/kg+via tail vein, iv, CTX solution 300 mg/kg), CTX + SchA low-dose, medium-dose, high-dose and excessive high-dose groups (ig, SchA 30, 300, 3 000, 30 000 μg/kg+via tail vein, iv, CTX solution 300 mg/kg) with 6 rats in each group. Blood samples were collected from orbital venous plexus of rats before medication and 0.083, 0.25, 0.5, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48 h after medication.UPLC-MS/MS method was applied for concentration determination of CTX and its metabolites [de-chloroethyl CTX (DC-CTX), 4-ketone CTX (4-keto CTX), carboxyl phosphamide (CPM)] in plasma of rats. The plasma concentration-time curve was obtained. The pharmacokinetic parameters were fitted by using DAS 2. 0 software. RESULTS: The maximum plasma concentration (cmax) of DC-CTX in CTX group, CTX+WZC group, CTX+SchA low-dose, medium-dose, high-dose and excessive high-dose groups were (22 167. 85 ±2 844. 93), (10 920. 53 ± 1 490. 89), (18 951. 29 ± 1 558. 81), (18 622. 08 ± 791. 19), (18 515. 20 ± 2 560. 61), (15 133. 21 ± 1 305. 07) μg/mL, respectively; the area under the curves (AUCo-48 h) were (173 864. 01 ± 65 342. 21), (100 996. 98 ± 33 530. 02), (137 028. 16 ± 45 975. 19), (131 650. 18 ± 53 196. 41), (113 699. 40 ± 34 131. 36), (110 773. 27 ± 30 307. 15) μg·mL/h, respectively. Compared with CTX group, cmax of DC-CTX in CTX group, CTX+SchA low-dose, medium-dose, high-dose and excessive high-dose groups were decreased by 50. 74％, 14. 51％, 16. 10％, 16. 48％, 31. 73％, respectively. AUC0-48 h were decreased by about 42. 23％, 21. 45％, 24. 63％, 33. 37％, 36. 55％, respectively; with statistical significance (P<0. 05). The pharmacokinetic indexes as t1/2, tmax had no significant change. CONCLUSIONS: To some degree, both WZC and SchA can reduce the generation of DC-CTX, which indicates both of them can inhibit CTX toxicity metabolism pathway so as to reduce the generation of toxic metabolite chloroacetaldehyde. The inhibitory effect of SchA on toxicity metabolism pathway is weaker than that of WZC, and does not have a dose-dependent inhibitory effect.

In recent years , many studies have found that vitamin D insufficiency is associated with osteomalacia , hyperten-sion, diabetes, metabolic syndrome and other diseases .Vitamin D must be hydroxylated in the liver by a 25-hydroxylase for the first time, and then in the kidney by a 1α-hydroxylase for the second time to form the active metabolite 1,25-dihydroxy vitamin D ,which binds to the intracellular vitamin D receptor and exerts its effects .This paper reviewed the relationship between vitamin D and disease , present research situation of gene polymorphism of vitamin D receptor , and the advantages and limitations of several methods of vitamin D detection, and proposed the best method for detecting vitamin D receptor gene polymorphism and the importance of the detection state to guide clinicians to use drug rationally .

Objective To establish the fingerprint spectrum and assay three active components (hesperidin, salvianolic acid B and chrysophanol) in Shenshuaining granule by HPLC method. Methods The chromatographic separation was achieved on SunFireTM C₁₈ column with acetonitrile-0.1% formic acid aqueous solution as mobile phase. Gradient elution program was applied with flow rate of 1.0 ml/min, detection wavelength at 254 nm and the column temperature at 25 ℃. The fingerprint spectrum was established and three active components in Shenshuaining granule were assayed. Results There were 22 common peaks on the fingerprints after analyzing chromatograms from 10 batches of Shenshuaining granules. Good fingerprint similarities (≥0.9) between different batches and the control chromatogram were found. This method has great repeatability, stability and precision, which meets all the assay requirements. Conclusion A simple and reliable HPLC method was developed, which is suitable for the fingerprint establishment of Shenshuaining granules. It provides a method for the quality control of Shenshuaining granules.

Objective To analyze the correlation of valproic acid and its metabolites (2-propyl-4-pentenoic acid, 3-hydroxy valproic acid,5-hydroxy valproic acid) with liver injury reference index. Methods 328 plasma samples from epilepsy patients were collected and divided into two groups（123 samples from patients with abnormal liver function, experimental group; 205 samples from patients with normal liver function, control group）.The plasma concentrations of valproic acid and its metabolites in the two groups were determined by LC-MS/MS method and the diagnostic value of the concentrations to liver disfunction was analyzed by ROC curve. Results The mean plasma concentration of valproic acid and its three metabolites in the patients with abnormal liver function was higher than that in the control group with was statistically difference（P<0.05）.The concentration of valproic acid and its metabolites could be used as a reference for the diagnosis of liver injury,5-hydroxy valproic acid had better diagnostic value than valproic acid. Conclusion The metabolites of valproic acid were associated with hepatotoxicity, which could be used as a diagnostic index of liver injury and could be a reference for clinical safe application of valproic acid.