Simultaneous determination of dextromethorphan, dextrorphan, and guaifenesin in human plasma using semi-automated liquid/liquid extraction and gradient liquid chromatography tandem mass spectrometry.

A method for the simultaneous determination of dextromethorphan (DEX), dextrorphan (DET), and guaifenesin (GG) in human plasma was developed, validated, and applied to determine plasma concentrations of these compounds in samples from six clinical pharmacokinetic (PK) studies. Semi-automated liquid handling systems were used to perform the majority of the sample manipulation including liquid/liquid extraction (LLE) of the analytes from human plasma. Stable-isotope-labeled analogues were utilized as internal standards (ISTDs) for each analyte to facilitate accurate and precise quantification. Extracts were analyzed using gradient liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Use of semi-automated LLE with LC-MS/MS proved to be a very rugged and reliable approach for analysis of more than 6200 clinical study samples. The lower limit of quantification was validated at 0.010, 0.010, and 1.0 ng/mL of plasma for DEX, DET, and GG, respectively. Accuracy and precision of quality control (QC) samples for all three analytes met FDA Guidance criteria of +/-15% for average QC accuracy with coefficients of variation less than 15%. Data from the thorough evaluation of the method during development, validation, and application are presented to characterize selectivity, linearity, over-range sample analysis, accuracy, precision, autosampler carry-over, ruggedness, extraction efficiency, ionization suppression, and stability. Pharmacokinetic data are also provided to illustrate improvements in systemic drug and metabolite concentration-time profiles that were achieved by formulation optimization.

Pharmacodynamic and pharmacokinetic effects in gingival crevicular fluid from re-dosing during brushing.

The IntelliClean System from Sonicare and Crest combines a rechargeable sonic power toothbrush and a novel liquid toothpaste into one integrated system, providing the opportunity to re-dose with toothpaste during the brushing cycle. The purpose of this study was to investigate cleaning effects from in-mouth re-dosing with toothpaste during the brushing cycle vs conventional bolus dosing. This was a randomized, examiner-blind, six-period, crossover clinical study. Eighteen adult subjects used an experimental integrated system employing either a re-dosing regimen (2 doses at the start of brushing with 1 additional in-mouth dose during the last 30 seconds of brushing [2+1]) or a conventional regimen (2 doses at the start of brushing only [2+0]). Gingival crevicular fluid (GCF) was sampled at the final brushing quadrant from a preselected site in the gingival sulcus using filter strips at baseline and at 4, 15, and 120 minutes postbrushing. Mean change from baseline in the concentrations of total facultative anaerobes (TFAs) and gram-negative anaerobes (GNAs) in the GCF at 120 minutes posttreatment were modeled separately using general linear mixed models. Area under the curve of surfactant (sodium dodecyl sulfate [SDS]) in GCF over 2 hours postbrushing was calculated and modeled using an analysis of variance model. All hypotheses were tested 2-sided at the 5% significance level. Relative to the conventional regimen, the re-dosing (2+1) regimen produced a significantly greater reduction in log10 (TFA colony-forming units [CFU]/microL GCF) after brushing, 0.99+/-0.12 vs 0.65+/-0.12 (mean change +/- standard error), and a significantly greater reduction in log10 (GNA CFU/microL GCF) after brushing, 0.75 +/-0.14 vs 0.45 +/- 0.14. The re-dosing regimen led to significantly more SDS in GCF relative to the conventional regimen over the 2-hour time period. Re-dosing of liquid toothpaste during the brushing cycle with the IntelliClean System leads to a significantly increased cleaning effect, as defined by a reduced bacterial count in GCF, and significantly higher levels of surfactant in the GCF up to 2 hours after the brushing event.

Semi-automated liquid--liquid back-extraction in a 96-well format to decrease sample preparation time for the determination of dextromethorphan and dextrorphan in human plasma.

A semi-automated, 96-well based liquid-liquid back-extraction (LLE) procedure was developed and used for sample preparation of dextromethorphan (DEX), an active ingredient in many over-the-counter cough formulations, and dextrorphan (DOR), an active metabolite of DEX, in human plasma. The plasma extracts were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS-MS). The analytes were isolated from human plasma using an initial ether extraction, followed by a back extraction from the ether into a small volume of acidified water. The acidified water isolated from the back extraction was analyzed directly by LC-MS-MS, eliminating the need for a dry down step. A liquid handling system was utilized for all aspects of liquid transfers during the LLE procedure including the transfer of samples from individual tubes into a 96-well format, preparation of standards, addition of internal standard and the addition and transfer of the extraction solvents. The semi-automated, 96-well based LLE procedure reduced sample preparation time by a factor of four versus a comparable manually performed LLE procedure.