| A sensitive,simple,and accurate HPLC method was developed for the determination of Adriamycin in rat plasma in the Section 1.Using daunorubicin as internal standard(I.S),Adriamycin in plasma was determined by HPLC with liquid-liquid extraction of the compound from plasma into organic solvent and was separated by the column of Phenomenex?ODS(250mm×4.6mm,5um) at 30°C.The mobile phase consisted of a mixture l0mmol/L potassium dihydrogen phosphate solution(adjusted to pH 3.0 with phosphoric acid)-acetonitrile(69:31,V/V) pumped at a flow rate of l.0mL/min.A fluorescence detector was set at an excitation wavelength of 490nm and an emission wavelength of 550nm.The drug-free plasma did not interfere with the determination of Adriamycin and I.S.There were good linear relationships(1/C2 weighted,r=0.9998) between peak area ratio of Adriamycin to I.S and C within the range of 0.025-2.0μg·mL-1.The lower limit of quantification was 0.0lμg·mL-1.The precision of inter-day and intra-day was less than 11%. Adriamycin in plasma was stable for at least 14 days and three freeze-thaw cycles. The method established in the paper can be applied to the clinical pharmacokinetics study.In Section 2,the high performance liquid chromatography(HPLC) method measuring Adriamycin concentration in plasma was used to measure and compare the pharmacokinetic parameters among three groups,which was administrated with the Adriamycin,Adriamycin and Danshen Tablet,Adriamycin and Salvia miltiorrhiza Injection respectivly.The pharmacokinetics parameters were calculated as followed. k21(h-1)were 0.095±0.026 of Adriamycin Group,0.17±0.040 of Adriamycin and Danshen Tablet Group,and 0.11±0.029 of Adriamycin and Salvia miltiorrhiza Injection Group;kl0(h-1) were 2.27±1.58,0.92±0.24,4.36±0.46 of the three groups above respectively;kl2(h-1) were 4.62±1.20,6.27±0.81,3.42±1.35;t1/2(a)(h) were 0.10±0.020,0.096±0.01,0.089±0.011;t1/2(β)(h) were 29.34±12.51,35.52±4.97, 12.02±1.81;Cl(L·h-1) were 0.67±0.14,1.25±0.56,0.71±0.041;Vss(L) were 26.65±6.09,62.09±24.69,12.39±2.29;AUC0-t(h·μg·L-1) were 1.49±0.26,0.58±0.27, 2.02±0.19;MRT(h) were 30.37±18.79,49.56±4.62,7.44±1.90.A sensitive,simple,and accurate HPLC method was developed for the determination of Adriamycin,Protocatechualdehyde and TanshinoneⅡA in rat plasma in the Section 3.Using daunorubicin as internal standard(I.S),Adriamycin in plasma was determined by HPLC with liquid-liquid extraction of the compound from plasma into organic solvent and was separated by the column of Phenomenex?ODS(250mm×4.6mm,5μm) at 30°C.The mobile phase consisted of a mixture l0mmol/L potassium dihydrogen phosphate solution(adjusted to pH 3.0 with phosphoric acid)-acetonitrile by a programming elution pumped at a flow rate of 1.0mL/min.A fluorescence detector was set at an excitation wavelength of 316nm and an emission wavelength of 414nm to detect TanshinoneⅡA.And a ultraviolet detector was set at an wavelength of 230nm to detect Protocatechualdehyde.The drug-free plasma did not interfere with the determination of Adriamycin,Protocatechualdehyde and TanshinoneⅡA and I.S.There were good linear relationships(1/C2 weighted,r>0.99) between peak area ratio of Adriamycin,Protocatechualdehyde and TanshinoneⅡA to I.S and C within the range of 0.01-1.0μg·mL-1.The lower limit of quantification was 0.01μg·mL-1.The precision of inter-day and intra-day was less than 11%.The method established in the paper can be applied to the clinical pharmacokinetics study.In section 4,firstly,the effect of Danshen to the toxicity of Adriamycin was studied.60 mice were randomly divided into 6 groups which were 3 negative control group and 3 adriamycin groups.The 3 adriamycin groups were given adriamycin through ip at the dosage of 15.00mgkg-1,and two of which were given Danshen Tablet,Sal via miltiorrhiza Injection respectively,then we observed the survival status in the following seven days.Finally,the animals death rates of adriamycin group was 60%,the ones of Danshen Tablet,Salvia miltiorrhiza Injection group were 70%and 10%respectively.Secondly,MTT assay was adopted to determine the in-vitro anti-tumor effect.The results suggested that the Salvia miltiorrhiza Injection may protected the cells from the toxicity of Adriamycin,and TanshinoneⅡA may promote the inhibit tumor effect of Adriamycin.
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