Assessment of dim light melatonin onset based on plasma and saliva samples.

Melatonin (MELA) is a nocturnal hormone involved in the regulation of the circadian rhythm. MELA can be detected in plasma and saliva, and its salivary concentration strongly correlates with its plasma concentration. Dim light melatonin onset (DLMO) is considered to be the most accurate objective marker for assessing the circadian phase. The purpose of the study was to establish a method for the determination of MELA in plasma and saliva based on the liquid chromatography with tandem mass spectrometry (LC-MS/MS) and compare DLMO using both plasma and saliva matrices. The validation of the LC-MS/MS methods was performed in accordance with the European Medicines Agency (EMA) guideline. The study was conducted on a group of 21 volunteers, male and females, aged 26-54 years. Plasma and saliva were collected at five time points: between 20:00 and 00:00 hours. The MELA concentration was determined by the LC-MS/MS. The DLMO was considered as the point in time when MELA concentration exceeds 20 pg/mL in plasma and 7 pg/mL in saliva. The correlation coefficient between the plasma and salivary MELA concentration was r = 0.764 (p < .001). The ratio of the plasma/saliva MELA concentrations was 2.87. The mean time of the DLMO in the plasma was 21:30 ± 0:45 hours, and in the saliva was as follows: 21:34 ± 1:00 hours. The correlation between the DLMO, calculated based on the plasma and saliva MELA profiles, was r = 0.679 (p < .05). The determination of salivary MELA concentration using LC-MS/MS allows for the determination of the DLMO. Our method may be applied in clinical practice for the diagnosis and monitoring of circadian rhythm disorders.Abbreviations: CE: Collision Energy; CID: Collision-Induced Dissociation; DL: Desolvation Module; DLMO: Dim Light Melatonin Onset; EFSA: European Food Safety Authority; EMA: European Medicines Agency; ESI: electrospray ionization; HB: heat block; HPLC: high performance liquid chromatography; IS: internal standard; K3EDTA: ethylenediaminetetraacetic acid tripotassium salt; LC-MS/MS: liquid chromatography with tandem mass spectrometry; LLE: liquid-liquid extraction; LLOQ: lower limit of quantification; MELA: melatonin; MELA-D4: melatonin-d4; MRM: multiple reaction monitoring; Q1: quadrupole 1; Q3: quadrupole 3; RE: relative error; RIA: radioimmunoassay; RSD: relative standard deviation; SD: standard deviation; ULOQ: upper limit of quantification.

Biorelevant In Vitro Release Testing and In Vivo Study of Extended-Release Niacin Hydrophilic Matrix Tablets.

Niacin (nicotinic acid, NA) is administered orally as an antihyperlipidemic agent in extended-release (ER) tablets in high doses. Due to rapid absorption and extensive metabolism (non-linear pharmacokinetics), the drug plasma levels are highly variable, which may correlate with side effects. Interestingly, this erratic drug delivery behavior of niacin ER products cannot be clarified by compendial in vitro release testing. The standard dissolution tests do not allow to mimic the selected GI tract characteristics in order to estimate the robustness of formulation under the variability of the physiological conditions. These are characterized by the pH value, impact of motility forces and composition, as well as volume of GI liquids. Our paper demonstrates a comparison of a newly developed ER HPMC niacin formulation with an originator product. The research aimed to design a robust matrix tablet of comparable biopharmaceutical behavior, safety and efficacy. The extensive in vitro investigation, including dynamic studies in flow-through cell apparatus and stress test device, forms the basis for the evaluation of nicotinic acid plasma concentrations in vivo. The occurrence of erratic, multiple NA plasma peaks after the administration of both extended-release products is a result of its local input excess over the metabolic threshold (at the level corresponding to maximum 2% of the administered dose, i.e., 20 mg of drug) due to the mechanical stresses of physiological intensity. We demonstrate how this behavior is similar for both marketed and test products. In this context, we describe how a robust ER matrix and well-designed formulation does not guarantee the test product's bioequivalence to the comparator one out of reasons unrelated to technology and biopharmaceutical properties, but because of the active compound's intrinsic pharmacokinetic characteristics, i.e., highly variable, extensive metabolism of nicotinic acid.

Feasibility of the Ph.Eur. flow-through cell for dissolution testing of the compounded rectal suppositories containing indomethacin or sodium diclofenac.

Ph.Eur. and BP have introduced a dissolution apparatus for suppositories. Suitability of the apparatus for quality control of indomethacin or sodium diclofenac (100 mg) compounded suppositories was evaluated and the effect of the type of suppository base on dissolution profiles was studied. The fastest and most reproducible release profiles were observed for hydrophilic base (macrogols). More than 80% of the drug was released during 60 min, while after 350 min 18.5-50% of the total amount was released from lipophilic bases (Witepsol and Adeps solidus). The results demonstrate that slow and non-reproducible release occurs when the lipophilic suppository base does not melt. The feasibility of the test for the formulations, which do not melt during the procedure, is questionable.