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Ivabradine French Servier company first developed antianginal new drugs, numerous studies have confirmed its safety and effectiveness, the European drug trial approved by the Board in 2005 11 menstrual listed 27 countries in Europe, for the treatment of β-blockers contraindicated or intolerant chronic stable angina. The drug is the first to select specific cardiac pacing current (If) inhibitors play alleviating the sinus node pacemaker activity speed, compared with traditional medicines, the mechanism does not affect blood pressure, hemodynamic myocardial contractility, atrioventricular or ventricular repolarization. Objective: domestic literature is not about hydrochloric ivabradine tablets of human pharmacokinetics reported, this study intends to explore and establish LC-MS/MS method (high performance liquid chromatography - tandem mass spectrometry) detected in human plasma in Iraq Laval to Bray given method, and the method is applied to the study of single and multiple doses ivabradine pharmacokinetic characteristics in healthy humans, first elucidated by hydrochloric ivabradine tablets in Chinese healthy try Chinese medicine pharmacokinetic characteristics of drugs in the human body absorption, distribution, metabolism and excretion of the dynamic changes in the characteristics and clarify its disposal law hydrochloric ivabradine given piece of the Phase II clinical trial study design and dosing scheme determined to provide a reference, to provide a theoretical basis for its clinical application. Methods: In this study, the following three parts, the first part of establishing good specificity, high sensitivity, reproducibility, the accuracy good LC-MS/MS detection methods. Detection steps: 1 mL, 10 mL centrifuge tube, precision plus blank plasma sample internal standard solution (2? G · mL -1 ) 50? L, and vortex 30 s mix. 4 mL of ethyl acetate, whirlpool 3 min, -1 centrifugation for 10 min at 4000 r · min . Absorb organic layer to another clean tube and blowing a nitrogen stream at 30 ° C in a water bath to near dryness. Residue 200? L mobile phase at 16000 r · min -1 centrifugation for 8 min, the supernatant is transferred to the autosampler sample tube, the injection volume 5μL by LC- MS / MS analysis. Hydrochloric acid after the second part of the study a single administration of ivabradine tablets in healthy subjects in vivo pharmacokinetic characteristics. 30 healthy subjects randomized parallel divided into three groups, each group of 10 healthy subjects (evenly divided between men and women), low, medium, high three doses of single oral dose of 2.5 mg, 5.0 mg, 7.5 mg hydrochloride Iraq Laval Bray given tablets, blood collection point: before administration, after administration of 8,15,30,45 min and 1,1.5,2,3,4,6,9,12,24,36,48 h, were collected blood 4 mL, opposed heparin tubes, centrifuged plasma set ordinary dry test tube and stored at -20 ℃, under test. LC-MS/MS method for determination of plasma ivabradine concentration DAS2.0 software to calculate the pharmacokinetic parameters. Of ivabradine after repeated administration of the third part of the research hydrochloride tablets in healthy subjects in vivo pharmacokinetic characteristics. Multiple dosing test is performed after the end of the single dose test, a single administration of low, medium, and high dose group continued administration, each dose is 2.5 mg, 5.0 mg, 7.5 mg, day 2 times for consecutive seven days. Multiple doses of blood collection: 4 to 7 days prior to administration in the morning, seven days after the administration 8,15,30,45 min and 1,1.5,2,3,4,6,9,12- 24,36,48 h blood 4 mL, placed in heparinized tubes, centrifuged plasma at -20 ℃ storage under test. LC-MS/MS method for determination of hydrochloric acid in the plasma ivabradine concentration, with DAS2.0 software calculate the pharmacokinetic parameters. Results: 1. Established in the test plasma ivabradine determination method specificity, high sensitivity, repeatability and good accuracy; ivabradine standard curve was linear over the range of 0.101 -1 01 ng · mL -1 , a good linear relationship; minimum detection limit of 0.101 ng · mL -1 ; batch of high, medium, and low concentration within and inter-assay precision RSD were less than 15.0%; extraction recoveries of the sample and the internal standard in more than 70%; medium effect measurement results; Stability of which adhere to the determination of biological samples. A ivabradine linear range of the standard curve 0.085-25.5 ng · mL -1 , a good linear relationship; minimum detection limit of 0.085 ng · mL -1 ; the high, medium, and low concentration intra and inter-assay precision RSD were less than 15.0%; samples and internal standard extraction recovery medium effect measurement results; stability study and adhere to biological samples in more than 70%; Determination of requirements. 2.30 healthy subjects a single administration of hydrochloric acid ivabradine tablets (2.5 mg, 5.0 mg, 7.5 mg), and its in vivo Iraq Laval Bray given pharmacokinetic parameters were: T max : 0.85 ± 0.43 h, 1.15 ± 0.24h, 1.00 ± 0.46 h; C max : 14.92 ± 4.05 ng · mL -1 , 35.98 ± 8.68 ng · mL -1 , 48.62 ± 9.62 ng · mL -1 ; t 1/2 : 3.87 ± 1.30 h, 5.03 ± 1.19 h, 5.00 ± 1.84 h; AUC 0-t : 64.23 ± 13.87 ng · mL -1 , 169.60 ± 33.09 ng · mL -1 , 215.92 ± 57.21 ng · mL -1 ; AUC 0 - ∞ : 64.88 ± 13.97 ng · mL -1 , 171.19 ± 33.45 ng · mL -1 , 217.95 ± 56.42 ng · mL -1 ; CLZ / F: 0.04 ± 0.01,0.03 ± 0.01 L · h -1 , 0.04 ± 0.01 L · h -1 ; V Z / F: 0.23 ± 0.09 L, 0.21 ± 0.03 L, 0.26 ± 0.10 L. A ivabradine pharmacokinetic parameters were: the T max : 1.08 ± 0.75 h, 1.30 ± 0.26 h, 1.13 ± 0.48 h; the C max : 1.81 ± 0.73 ng · mL -1 , 3.11 ± 1.61 ng · mL -1 , 6.26 ± 1.02 ng · mL -1 ; t 1/2 : 8.15 ± 3.09 h, 9.64 ± 1.32 h, 10.78 ± 2.67 h; AUC 0-t : 13.28 ± 3.26 ng · mL -1 , 23.33 ± 9.17 ng · mL -1 , 43.94 ± 9.22 ng · mL -1 ; AUC 0 - ∞ : 15.04 ± 3.09 ng · mL -1 , 25.39 ± 9.49 ng · mL -1 , 46.87 ± 10.05 ng · mL -1 ; CL Z < / sub> / F: 0.17 ± 0.04 L · h -1 , 0.24 ± 0.13 L · h -1 , 0.17 ± 0.03 L · h -1 < / sup>; Vz / F: 2.11 ± 1.31 L, 3.22 ± 1.54 L, 2.50 ± 0.38 L. 3.30 in healthy subjects given hydrochloric Iraq repeatedly cutting Bray given tablets (2.5 mg, 5.0 mg, 7.5 mg), Iraq Laval Bray given pharmacokinetic parameters: T max : 0.85 ± 0.44 h, 1.08 ± 0.53 h, 1.05 ± 1.05 h; C max : 18.32 ± 3.27 ng · mL -1 , 37.88 ± 10.56 ng · mL -1 , 50.01 ± 17.46 ng · mL -1 ; t 1/2 : 4.55 ± 1.62 h, 6.68 ± 2.05 h, 7.15 ± 1.90 h ; AUC (0-48) : 93.40 ± 22.86 ng · mL -1 , 209.89 ± 71.26 ng · mL -1 , 326.49 ± 135.05 ng · mL -1 ; AUC ss : 79.78 ± 16.87 ng · mL -1 , 165.61 ± 50.30 ng · mL -1 , 243.61 ± 87.08 ng · mL -1 ; CL / F: 0.03 ± 0.01 L · h -1 , 0.03 ± 0.01 L · h -1 , 0.03 ± 0.01 L · h -1 ; V d / F: 0.18 ± 0.06 L, 0.25 ± 0.09 L, 0.27 ± 0.15 L. Norepinephrine after repeated administration of ivabradine pharmacokinetic parameters: T max : 0.83 ± 0.47 h, 1.23 ± 0.80 h, 1.30 ± 1.43 h; C max < / sub>: 2.60 ± 0.83 ng · mL -1 , 4.94 ± 1.76 ng · mL -1 , 7.59 ± 1.79 ng · mL -1 ; t 1/2 : 11.47 ± 2.83 h, 14.25 ± 2.44 h, 13.79 ± 3.23 h; AUC (0-48) : 30.15 ± 9.77 ng · mL < sup> -1 , 58.05 ± 19.92 ng · mL -1 , 102.19 ± 20.72 ng · mL -1 ; AUCss: 17.51 ??± 5.11 ng · mL < sup> -1 , 30.54 ± 10.21 ng · mL -1 , 52.83 ± 12.94 ng · mL -1 ; CL / F: 0.09 ± 0.05 L · h -1 , 0.09 ± 0.05 L · h -1 , 0.07 ± 0.01 L · h -1 ; V d / F: 1.33 ± 0.30 L, 1.83 ± 0.76 L, 1.37 ± 0.34 L; accumulation factors of low-medium-high three dose groups were: 1.44 ± 0.22,1.21 ± 0.37,1.51 ± 0.59. Conclusion: 1 The experimental set up human plasma ivabradine given and norepinephrine Iraq Laval Bray given LC-MS/MS detection method precision, accuracy, selectivity, good reproducibility, applicable to human plasma ivabradine and norepinephrine ivabradine pharmacokinetics dynamics study. Single administration of low, medium and high dose groups plasma ivabradine and norepinephrine ivabradine C max and AUC0-t linearly with dose-related, indicating The ivabradine piece was linear pharmacokinetic characteristics within 2.5-7.5 mg. .3. The multiple doses Iraq cutting Bray given steady state the pharmacokinetic parameter T max t 1/2 and pharmacokinetics of single dose The kinetic parameters are basically the same the ivabradine and norepinephrine ivabradine no accumulation in the body. Hydrochloric acid under this dose of ivabradine tablets have good security.
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