Objective: To develop the method for the simultaneous analysis of cyclophosphamide and 4-hydroxycyclophosphamide (4-OHCP) in Dried Blood Spot (DBS) using Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) and its application in breast cancer patients for therapeutic drug monitoring. Methods: Sample preparation used protein precipitation with methanol and acetonitrile (2:1 v/v). The separation was conducted using 1.7μm (2.1 x 100 mm) Waters AcquityTM UPLC C18 column; mobile phase consists of 0.01% formic acid and methanol (50:50 v/v) with isocratic elution, column temperature 30 °C, flow rate 0.3 ml/min and hexamethylphosphoramide (HMP) used as an internal standard. Analysis was performed by a triple quadrupole mass spectrometry with a positive ion mode of Electrospray Ionization. Cyclophosphamide was detected at m/z 260.968>139.978, 4-OHCP at m/z 338.011>224.979, and HMP at m/z 180.17>92.08. The method was applied to quantify cyclophosphamide and 4-OHCP in DBS of breast cancer patients. Blood samples were collected at 2 and 4 h after cyclophosphamide administration for therapeutic drug monitoring. Results: The method was linear in the range of 50–30.000 ng/ml for cyclophosphamide and 10–1000 ng/ml for 4-OHCP. Lower Limit of Quantification (LLOQ) concentration of cyclophosphamide was 50 ng/ml and 4-OHCP was 10 ng/ml. Accuracy and precision within-run and between-run met the requirements with % diff and CV, not exceeding ±15% and not more than ±20% for LLOQ concentration. The results from DBS samples of cancer patients showed that the level of cyclophosphamide was in the range of 6045.980 ng/ml to 37024.403 ng/ml and 4-OHCP was in the range 33.155 ng/ml to 246.362 ng/ml. Conclusion: The developed method met the requirements of all validation parameters under the Guideline on Bioanalytical Method Validation by the European Medicines Agency in 2011. Method can be applied on DBS of cancer patients and the results showed that cyclophosphamide and 4-OHCP was detected on 17 samples of breast cancer patients. This can be one of the parameters for therapeutic drug monitoring.

Objective: This study aimed to obtain the physicochemical properties of hydroxypropyl cellulose (HPC) powder from α-cellulose Betung bamboo and its characteristics in tablet formulation.Methods: HPC was prepared by hydroxypropylation of α-cellulose using 25% (w/v) sodium hydroxide and 10 ml propylene oxide (based on 1 g αcellulose) at 70 °C for 3 h. HPC of Betung bamboo (HPC BB) was characterized using fourier transform infrared (FTIR) spectrometry, particle size analyzer (PSA), x-ray diffraction (XRD), scanning electron microscope (SEM) and compared to HPC grade SL (HPC SL) as the reference. Then, HPC BB was used as a binder in tablet formulation by direct compression method and the resulted tablets were evaluated. The tablets evaluation including weight and size uniformity, hardness, friability and disintegration time. Results:The results showed HPC BB powder was yellowish white, odorless and tasteless, pH 7.49, residue on ignition 0.68%, hydroxypropoxy groups content 54.75%, average particle size 37.39 μm, loss on drying 1.09%, and moisture content 3.34%. Flow properties of powder fulfilled the requirements based on literature. Infrared spectrum and diffraction pattern of HPC BB were relatively similar to HPC SL. The tablets have average weight 403.495 mg, diameter 12.16 mm, thickness 3.11 mm, hardness 4.11 KPa, friability 2.04% and disintegration time 24.88 s. Conclusion:Based on the comparison of powder characteristics and tablets evaluation, HPC BB has a great potential in tablet formulation which showed similar characteristics to reference. I In nt te er rn na at ti io on na al l J Jo ou ur rn na al l o of f A Ap pp pl li ie ed d P Ph ha ar rm ma ac ce eu ut ti ic cs s

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Xylanase (endoxylanase, EC 3.2.1.8) is a commercial enzyme that has been applied in the industrial production of fuel, food, textiles and paper. Xylanase was isolated from the culture supernatant of Bacillus sp. AQ-1 grown on Nakamura medium containing 0.5% powder bunch palm oil. The optimum pH and temperature of xylanase activity were pH 7.0 and 60 °C, respectively. The enzyme was purified by anion exchange chromatography using DEAE-Sepharose-Fast-Flow column and gel filtration chromatography using Sephacryl S-300 column. The results showed that purification of xylanase produced two forms of xylanase, which were identified as xylanase A and xylanase B. Xylanase A can be separated from xylanase B by ultrafiltration using a 30 kDa polyethersulfone membrane. The molecular weight of xylanase A and B were 15.7 and 57.7 kDa, respectively.

Objective: This study aimed to design and optimize a gas chromatography-flame ionization detection (GC-FID) method to determine the bisphenol A (BPA) content in Indonesian canned food samples.Methods: GC with Hewlett-Packard-1 capillary columns (length, 30 m; inside diameter, 0.25 mm; and film thickness, 0.25 µm) was used with a column temperature of 150°C that was programmed to increase by 10°C/min to 260°C. Injector and detector temperatures were 280 and 300°C, respectively, the gas flow rate was 1.0 mL/min, and injection volume was 3.0 µL. Three types of canned food samples were prepared by ethyl acetate extraction and stored under four different conditions (4-8°C, 25-30°C, 40°C for 30 min, and 40°C for 60 min) to determine BPA migration levels.Results: Method validation (system compatibility, selectivity, calibration curve linearity, accuracy, and precision) was acceptable for BPA concentrations ranging from 2 to 15 μg/mL, with a coefficient of correlation of 0.99983. The limits of detection and quantitation were 0.287 and 0.956 μg/mL, respectively. Only one canned food sample type (Group A) showed BPA contamination under all storage conditions and exceeded the recommended guidelines for daily ingestion. Conclusion:The optimized GC-FID method was selective and relatively sensitive in the detection and quantitation of BPA. Furthermore, higher storage temperatures and durations increased the level of BPA migration into food.

Objective: To obtain an optimum and validated method for analyzing lercanidipine in plasma using Ultra Performance Liquid Chromatography of Tandem Mass Spectrometry (UPLC-MS/MS). Methods:The separation was carried out using 1.7μm (2.1 x 100 mm) Waters Acquity TM UPLC C18 column, a mobile phase of the 0.1% formic acidmethanol mixture (20:80 v/v) with isocratic elution, 30 °C column temperature, 0.2 ml/min flow rate and amlodipine as an internal standard. Mass detection was performed with a positive XBL TQD type Electrospray Ionization (ESI) in Multiple Reaction Monitoring modes. Lercanidipine was detected at m/z value of 612.11>280.27 and amlodipine was detected at m/z value 409.1>238.15. The optimum sample preparation method was a liquid-liquid extraction using 5 ml of n-hexane-ethyl acetate (50:50 v/v), vortex mixed for 3 min, centrifuged at 4000 rpm for 20 min, evaporated with nitrogen at 50 °C for 30 min, and the residue was reconstituted with 100 μl of mobile phase. Results:The method was linear in the range of 0.025-10 ng/ml with r ≥ 0.9986. Accuracy and precis ion within-run and between-run met the requirements with %diff and %CV, not exceeding ± 15% and not more than ± 20% for Lower Limit of Quantification (LLOQ) concentration. Conclusion:It was concluded that the developed method met the requirements of selectivity, carry over, stability, the integrity of dilution, and matrix effects under the Guideline on Bioanalytical Method Validation by the European Medicines Agency in 2011.