Simultaneous Determination of 1,4‐Benzodiazepines and Tricyclic Antidepressants in Saliva after Sequential SPE Elution by the Same HPLC Conditions

Abstract

A new method has been described to determine both benzodiazepines (six) and tricyclic antidepressants (four) simultaneously in saliva by HPLC with a UV detector set at 240 nm using cholchicine as the internal standard. A careful specific sequential solid-phase elution was optimized and performed to elute benzodiazepines using a mixture of methanol-acetonitrile (1:1 v/v) followed by the elution of tricyclic antidepressants with methanol. Separation of the compounds was performed on a Kromasil column (250 × 4 mm, 5 µm) by a gradient eluents consisting of 0.05 M CH 3 COONH 4 -acetonitrile-methanol (55:15:30 v/v/v). The results were linear for both benzodiazepines and tricyclic antidepressants up to 20 ng µL -1 with the correlation coefficients greater than 0.998. The sensitivity limits, LOD and LOQ were 0.08-0.34 ng µL -1 and 0.28-1.13 ng µL -1 , respectively. The method is simple, fast and reliable with good specificity and sensitivity, will be suitable for use in a clinical setting, where there is a concomitant use of 1,4-benzodiazepines and tricyclic antidepressants.

Electromembrane extraction‐high‐performance liquid chromatography‐ultraviolet detection of phenobarbital and phenytoin in human plasma, saliva, and urine

“…However, other biological matrices that is, saliva, dried blood spot, sweat, tear, cerebrospinal fluid, and hair have been employed as well. [6][7][8] Saliva as an alternative matrix for TDM, offers considerable benefits over serum, (a) salivary drug concentration indicates the unbound drug concentration in serum, (b) saliva sampling is a simple, noninvasive and inexpensive procedure which allows us to have repeated sampling. Patient can do the collection on his/her own.…”

An Overview on Total Analytical Methods for the Detection of 1,4-Benzodiazepines

“…There was no obvious preference for choice of column, except for reversed-phase C 18 [55] or C 8 , [56] which was more prevalent than the alternative phases. Monolithic columns [57], consisting of continuous beds with through pores of organic or inorganic matrix, have found increasing applications for HPLC separations.…”

HPLC and its Essential Role in the Analysis of Tricyclic Antidepressants in Biological Samples

“…The method was proved to be suitable for use both in clinical analysis, as well as for quality control of pharmaceutical formulations [64,65]. The analytes and the internal standard were extracted by Nexus (Varian) SPE cartridge and separated by 15 min, utilizing a Kromasil C 8 (150 × 4.6 mm; 5 mm) column with a gradient mobile phase containing methanol, acetonitrile, and 0.05 M ammonium acetate, delivered at a flow rate of 1.0 mL/min.…”