\n\nA simple and sensitive Ultra Performance Liquid Chromatography-Mass Spectrometry\nmethod was developed and validated to measure the concentrations of Alogliptin (ALO),\nLinagliptin (LIN), Saxagliptin (SAX), and Sitagliptin (SIT) using Pioglitazone (PIO) as an internal\nstandard.\n\n\n\nChromatographic separation of six gliptins was achieved on a C-18 column (100×2.1 mm,\n2.7 μm) using a mobile phase consisting of formic acid in water, 0.1%v/v: acetonitrile in gradient elution.\nElectrospray ionization (ESI) source was operated in the positive ion mode. Targeted MS/MS mode\non a QTOF MS was used to quantify the drug utilizing the transitions of 340.1(m/z), 473.2 (m/z), 316.2 (m/z),\n408.1 (m/z), and 357.1 (m/z) for ALO, LIN, SAX, SIT and PIO respectively.\n\n\n\nAs per ICH Q2R1 guidelines, a detailed validation of the method was carried out and the\nstandard curves were found to be linear over the concentration ranges of 1516.0-4548.1 ng mL-1, 519.8-\n1559.4 ng mL-1, 1531.4-4594.3 ng mL-1and 1519.6-4558.8 ng mL-1 for ALO, LIN, SAX and SIT respectively.\nPrecision and accuracy results were within the acceptable limits. The mean recovery was found to be\n98.8 _ 0.76 % (GEM), 102.2 _ 1.59 % (LIN), 95.3 _ 2.74 % (SAX) and 99.2 _ 1.75 % (SIT) respectively.\n\n\n\nThe optimized validated UPLC QTOF-MS/MS method offered the advantage of shorter\nanalytical times and higher sensitivity and selectivity. The optimized method is suitable for application\nin quantitative analysis of pharmaceutical dosage forms for QC laboratory.\n

Simultaneous Determination of Alogliptin, Linagliptin, Saxagliptin, and Sitagliptin in Bulk Drug and Formulation by UPLC Q-TOF-MS

A simple reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed and validated for simultaneous determination of alogliptin benzoate and pioglitazone HCl in bulk and tablet dosage form. Chromatographic analysis was performed on an Inertsil® ODS-3 column (250x4.6 mm, 5μm) with a mixture of methanol: phosphate buffer (pH 3) in the ratio 80:20 as mobile phase, at a flow rate of 1.0 mL min-1. UV detection was performed at 269 nm using DAD detector. The method was validated according to ICH guidelines. The retention times of alogliptin benzoate and pioglitazone HCl were 2.31±0.00 and 4.92±0.096 min, respectively. Calibration plots were linear over the concentration ranges 5-100 μg mL-1 for both drugs. The Limits of detection were170 and 215 ng mL-1and the quantification limit were 500 ngmL-1 and 650 ng mL-1 for alogliptin benzoate and pioglitazone HCl respectively. The accuracy of the proposed method was determined by recovery studies and was found to be 99.56% for alogliptinand101.38% for pioglitazone. The method was transferred to UPLC system using Syncronis®C18 column (50x 2.1 mm, 1.7 μm) and binary mixture was separated with good resolution within less than one minute.

https://www.imedpub.com/articles/development-and-validation-of-a-reversed-phase-hplc-method-forsimultaneous-determination-of-antidiabetic-drugs-alogliptinbenzoate.pdf

Development and validation of a reversed phase HPLC method forsimultaneous determination of antidiabetic drugs alogliptinbenzoate and pioglitazone HCl

Objective: A simple and stability-indicating high-performance thin-layer chromatographic method was developed and validated for the simultaneous estimation of metformin and alogliptin in tablets.
Methods: The method was developed in TLC aluminum plates pre-coated with silica gel 60F254 as the stationary phase and the solvent system consists of methanol: chloroform: 0.5% ammonium sulphate [4:4:2, v/v/v]. The system was found to conferred a compact spot for metformin [Rf value of 0.44±0.02] and alogliptin [Rf value of 0.66 ± 0.22]. Densitometric analysis of metformin and alogliptin was carried out at the wavelength of 254 nm. Forced degradation studies were conducted to know the stability of the drug samples under various stress conditions like acid, base, peroxide, photolytic degradation according to the ICH guidelines.
Results: The developed method was found to be suitable for the excellent separation of the drug samples. Calibration curves were linear in the range of 40-200 ng/spot with a correlation coefficient of 0.996 for metformin and calibration curves were linear in the range of 1-5 ng/spot with a correlation co-efficient of 0.997 for alogliptin, respectively. Stability study shows that the chromatograms of samples degraded with acid, base, hydrogen peroxide, dry heat, and photolytic showed well-separated spots of pure metformin and alogliptin as well as some additional peaks at different Rf values. The method was successively applied to pharmaceutical formulation. No chromatographic interference from the tablet excipients was found.
Conclusion: The newly developed method can be applied for the identification and quantitative determination of metformin and alogliptin in the combined dosage form.

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Development and validation of stability-indicating simultaneous estimation of metformin and alogliptin in tablets by high-performance thin layer chromatography

Sensitive LC-MS/MS (Liquid Chromatography Tandem Mass Spectrometric) Method for the Simultaneous Determination of Alogliptin and Voglibose in human plasma. A highly sophisticated and sensitive LC-MS/MS method has been developed and validated for the Alogliptin and Voglibose simultaneous determination in human plasma. Alogliptin D3 and Miglitol were used as IS (Internal standard). Protein precipitation extraction was followed for the analytes and IS. Chromatography conditions included an isocratic mobile phase composing of 5 mM Ammonium formate: Acetonitrile in the ratio 50:50 v/v. The column used was Welchrom XB C18, with specifications of 50 × 4.6 mm, 5 μm, at a flow rate of 0.70 ml/min. The retention time of Alogliptin, Voglibose, Alogliptin D3 and Miglitol occurred at ~1.03, 0.8, 0.8 and 0.81 min respectively and the total chromatographic run time was 3.0 min. Alogliptin and Voglibose achieved a linear response function in human plasma at 5.09-509 ng/mL and 2.03-203 ng/mL respectively. Alogliptin and Voglibose achieved an intra and inter-day accuracy and precision in the range of 0.94- 4.35 and 0.91-3.89%; 1.41-10.8 and 1.90-7.75% respectively. The method was strictly validated according to the ICH guidelines. The results obtained from this study can be significantly utilized for developing full pharmacokinetic profiling in individuals.

Hemavathi G

Papers

Sensitive LC-MS/MS Method for the Simultaneous Determination ofAlogliptin and Voglibose in Human Plasma