Materials

Reagents

Methaqualone Solvent - serum

Internal standard(PBCA) solvent - Chloroform

Alkaline buffer - Borate buffer (pH 10.5) (Aqueous Phase) - This buffer is prepared by
adding about 8ml of NaOH(6 mol/liter) to 100 ml of a saturated sodium borate solution which will yield a pH 10.5 borate buffer.


Why do we need this pH 10.5 buffer?

PBCA and methaqualone functioned best between pH 9.5-11.0. Also interferences from biological substances were less. Therefore, the authors decided to use a pH of 10.5.



Internal Standard: PBCA( N-Propionyl-2- benzoyl-4-chloro-aniline)

Analyte: Methaqualone (prepared from the pure standard of Methaqualone hydrochloride)

Concentrations of
5, 10, 20, 30, and 40 mg/liter were prepared <------------------------ These are the Standards

The serum standards were placed into individual 5-ml
vials, frozen, and stored at 4 degrees C until needed.

Apparatus:
F&M 402 gas chromatograph
dual hydrogen-flame detectors(FID)
A U-shaped glass column (150 cm, 2 mm i.d.)
was packed with “3% OV-1 (methyl silicone) on
100/120 mesh Gas Chrom Q”
flow rate for helium, hydrogen, and air were
30, 20, and 380 ml/min,
column temperature was 210  degree C; detector temperature was 300
degree C; flash-heater temperature was 310 degree c.
electrometer settings were: range X10 and attenuation X4. Strip-chart recorder (Hewlett-Packard
Model 7127A) speed was 1.27 cm/mm.