Determination Of Cefcapene In Human Plasma And The Pharmacokinetic Study Of Cefcapene Pivoxil Hydrochloride Granules In Healthy Volunteers

PURPOSEA quantitative analytical method of cefcapene in human plasma basedon high performance liquid chromatography tandem mass spectrometry wasdeveloped for the samples collected after10healthy volunteers was givenlow, middle, high and multiple doses of cefcapene pivoxil hydrochloridegranules. By interpreting the pharmacokinetic characteristics ofcefcapene following dosing of cefcapene pivoxil hydrochloride granules,statistics for later clinical experiments was achieved and foundationssupporting the clinical rational medication was built.METHODPlasma sample preparation by liquid-liquid extraction with ethylacetate was followed by the chromatography on a Zorbax TC-C18column（150mm×4.6mm i.d.,5μm） using acetonitrile-0.1%formic acid （25:75,v/v） at a flow rate of1.0mL/min and10μL was analyzed by LC-MS/MS.Using an electrospray source in the positive ion mode, multiple-reactionmonitoring was performed using the transitions m/z453.9�'207.9and m/z264.1�'58.0for cefcapene and internal standard （I.S.）ctahydroaminoacridine （OHA）, respectively.The validation of this analytical method was performed, includingselectivity, the range of standard curve, lower limit of quantification,accuracy, precision, recovery, matrix effect and stability.In this research, plasma samples from10healthy volunteers given with 50mg,100mg,200mg and multiple doses of cefcapene pivoxil hydrochloridegranules was analyzed in accordance with GCP requirements and the opinionsof the ethics committee。With the plasma concentrations achieved, thepharmacokinetic parameters of cefcapene was calculated using DAS3.0software and analyzed by SPSS16.0statistic analysis software.RESULTSThis LC-MS/MS method for the determination of cefcapene in humanplasma was sensitive, rapid, and strong enough. The calibration curve wasconstructed over the range0.025-7.5μg/mL with a lower limit ofquantitation of0.025μg/mL. The method only needs50μL plasma sampleand3.5min for each time. This analytical method is strong enough andthere is no potential interfering substances such as endogenous matrixcomponents or others. The accuracy （as relative error, RE） is less than±4.47%and the intra-and inter-day precisions （as relative standarddeviation, RSD） were less than15%. The recoveries of cefcapene plasmasamples of low, middle, high concentrations （0.075,0.75,6μg/mL） andOHA were90.33±6.86%,92.95±4.28%,85.88±2.44%and95.68±3.37%,respectively. The matrix effects of cefcapene plasma samples of low,middle, high concentrations （0.075,0.75,6μg/mL） and OHA were103.69±9.38%,97.68±13.01%,100.49±11.57%and97.89±6.92%,respectively. The method is stable enough and would not affected by matrixeffects. The concentrations of cefcapene remains stable when plasmasamples were stored at room temperature for1.5h, frozen at-80°C andthawed in room temperature for three times repeatedly, stored at roomtemperature for4h after treated, stored at-80°C for30days. Theconcentration of cefcapene and OHA would also remains stable at roomtemperature for4h and at-20℃for17h in stock solutions. The R.E.ofall the stability results were less than±15%, indicating all the sampleswere stable enough till the analyzation was finished. The average pharmacokinetic parameters of cefcapene in human bodyafter given with low, middle, high and multiple doses are as follows:Cmaxare0.61±0.18μg/mL,1.20±0.39μg/mL,1.76±0.79μg/mL and0.60±0.16μg/mL；t1/2z are1.42±0.32h,1.36±0.25h,1.45±0.36h and1.25±0.16h；Tmaxare2.15±0.53h,2.00±0.67h,3.00±0.97h and2.45±0.44h；AUC0-tare1.91±0.44μg/mL*h,3.80±1.08μg/mL*h,6.84±2.74μg/mL*h and1.92±0.49μg/mL*h；AUC0-∞are1.93±0.45μg/mL*h,3.84±1.10μg/mL*h,6.97±2.78μg/mL*h and1.93±0.49μg/mL*h；MRT are3.20±0.39h,3.04±0.36h,3.81±0.51h and2.97±0.29h；Vz/F are54.95±5.11L,53.23±12.63L,67.55±28.99L and49.84±14.80L；CLz/F are27.43±7.59L/h,27.70±6.66L/h,32.09±9.98L/h and27.76±8.69L/h。AUC_0-t、AUC_0-∞、Cmaxhas remarkable liner correlation to the dose, withthe correlation coefficient （r） and P values are0.779（P=0.000<0.01）、0.781（P=0.000<0.01）、0.676（P=0.000<0.01）, respectively. Within doses,t1/2z、MRT、Vz/F had no significant differences（P>0.05）, while CLz/F hadsignificant difference（P=0.001<0.01）. Tmaxshowed significant differenceonly between doses of100mg to200mg（P<0.01）. Between single and mulitpledoses, the main pharmacokinetic parameters including t1/2z、C_max、AUC_0-t、CLz/F、Vz/F had no significant differences（P>0.05）. And there was nosignificant differences between W1/-48、W2/-24、W3/0during multipledosing。...