Postmortem Analysis of Benzodiazepines in Human Bone by Gas Chromatography–Mass Spectrometry

A procedure based on gas chromatography-mass spectrometry was developed for the analysis of benzodiazepines (nordiazepam, oxazepam, lormetazepam, lorazepam, clonazepam, bromazepam and alprazolam) in postmortem human ribs. Powdered bone samples, including marrow remains inside, with the internal standard diazepam-d5 were subjected to enzymatic hydrolysis with 100 μL of β-glucoronidase and were incubated in sodium hydroxide for 1 h in a 70°C oven. Samples underwent liquid phase extraction and ethyl acetate was used as eluent. Chromatography was performed on a fused silica capillary column and the selected-ion-monitoring mode was used for analytes determination. The method was validated in the range 0.1–0.5 ng/mg (depending on the benzodiazepine) to 100 ng/mg with average values of recovery, matrix effect and process efficiency ranged from 83.2 to 94.3%, from 97.3 to 102.1% and from 80.5 to 91.2%, respectively. The intra- and inter-day accuracy was <15%. The procedure was tested in rib specimens obtained during routine autopsies from 20 cases where these benzodiazepines were found in blood. Benzodiazepines were detected in the combined bone and marrow samples in 60% of cases. Lorazepam was detected in bone in the range of 0.3–0.7 ng/mg, nordiazepam at 1.3–4.2 ng/mg and oxazepam at 1.1–1.2 ng/mg. To our knowledge, this protocol for the simultaneous analysis of these benzodiazepines is the first performed and validated using human ribs.

Determination of Ferulic Acid in “AI-FU-NUAN-GONG” Pills by High Performance Capillary Electrophoresis

High performance capillary zone electrophoresis (HPCE) was used in this text to determine the content of ferulic acid in a ready-made Chinese herbal medicine called AI-FUNUAN-GONG (in pills). Electrophoretic separation conditions: Uncoated fused silica capillary column (75um×49/58cm), injection height 7.5cm, 30mmolL-1 borax solution, separation voltage 20kV, injection time 10s, detection wavelength 315 nm, experimental temperature 20°C.Linearity was kept in the concentration ranging from 1~62.5 mg·L-1 of ferulic acid with correlation coefficient of 0.970. The average recovery was 97.541% and RSD value was 6.57%. Sample content of ferulic acid was 0.0715mg/g. This method has the advantages of strong specificity, simpleness, speediness, accurate results and good repeatability in separation and it is useful for determination of ferulic Acid in AI-FU-NUAN-GONG Pills.

Determination of Ferulic Acid in“SHI-QUAN-DA-BU” Pills by High Performance Capillary Electrophoresis

High performance capillary electrophoresis (HPCE) was used in this text to determine the content of ferulic acid in “SHI-QUAN-DA-BU” Pills. Electrophoretic separation conditions: Uncoated fused silica capillary column(75um×49/58cm), injection height 7.5cm, 30mmol•L-1borax solution, separation voltage 20kV, injection time 15s, detection wavelength 315 nm, experimental temperature 20°C. Linearity was kept in the concentration ranging from 1~31.25 mg·L-1of ferulic acid with correlation coefficient of 0.970. The average recovery was 97.56% and RSD value was 6.4%. The content of ferulic acid was 0.0855 mg/g (n=6).