Stock solution stability was proved for 9 days and evaluated. Stability of the drug in plasma samples was proved at LQC, HQC levels using six replicates each with its freshly prepared samples of same concentration. Reinjection reproducibility stability, benchtop stability, autosampler stability, freeze–thaw stability and long term stability was proved for drug in plasma samples. The reinjection

reproducibility was evaluated by comparing the extracted plasma samples that were injected immediately (time 0 h), with the samples that were re-injected after storing in the click here autosampler at 4 °C for 26 h. Stability samples were kept on bench (Benchtop stability) for 25 h and processed along with freshly prepared standards and proved the stability for 25 h. The stability of spiked human plasma samples prepared and stored at 4 °C in autosampler (autosampler stability) was evaluated for 79 h. Freeze–thaw stability at −30 °C at 4th cycle was performed and proved for 3 cycles by comparing with freshly prepared samples. Long term stability was proved for 34 days with its freshly prepared standards at respective concentrations. All these stability samples % Accuracy was less than 15%. The stability was proved as per USFDA guidelines.13 The bioanalytical method described above was applied to determine acamprosate concentrations in plasma following oral administration INCB024360 supplier of healthy human volunteers. These volunteers were contracted in APL Research

centre, Hyderabad, India and to each one of the 14 healthy volunteers were administered

a 333 mg dose (one 333 mg tablet) via oral with 240 ml of drinking water. The reference product CAMPRAL® tablets, Manufactured by Forest pharmaceuticals, INC. USA. 333 mg, and test product Acamprosate tablet (test tablet) 333 mg were used. Study protocol was approved by IEC (Institutional Ethical committee) and by DCGI (Drug Control General of India). Blood samples were collected as pre-dose (0) hr 5 min prior to dosing followed by further samples at 0, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.75, 6.5, 7.25, 8, 9.5, 12, 14, 18, 24, 30, 36, 48, 56, 60, 72, 84 and 96 h. After dosing, 5 ml blood sample was collected each pre-established time in vacutainers containing K2EDTA. A total of 50 (25 time points for reference, and 25 for test) time points were collected and centrifuged at 3200 rpm, 10 °C, 10 min. Then they were kept frozen at −30 °C until sample analysis. Oxalosuccinic acid Test and reference were administered to same human volunteers under fasting conditions separately and these volunteers were washed minimum 9 days intervals as per protocol approved by IEC. Pharmacokinetics parameters from human plasma samples were calculated by a non-compartmental statistics model using WinNon-Lin5.0 software (Pharsight, USA). Blood samples were taken for a period of 3–5 times the terminal elimination half-life (t1/2) and it was considered as the area under the concentration time curve (AUC) ratio higher than 80% as per the FDA guidelines.