22.1.02
Preparation of GC Column
AOAC Official Method 978.29
Cocaine Hydrochloride in Drug Powders
Gas Chromatographic Method
First Action 1978
Final Action 1983
A. Principle
Cocaine is extracted from weakly basic, aqueous solution with
CHCl3 containing internal standard, and then separated by GC from
other amines, organic-soluble neutrals, and internal standard.
B. Apparatus and Reagents
(a) Gas chromatograph.—With flame ionization detector and
1.8 m (6 ft) × 4 mm (id) glass column packed with 3% OV-1 on
100–120 mesh Chromosorb W (flux-calcined, acid washed, silanized,
diatomaceous earth). Typical operating conditions: temperatures
(°C): column 225, detector and injector 240; N2 carrier gas flow rate,
60 mL/min. Adjust column temperature to elute cocaine in 3 ±
0.5 min. Adjust H2 and air flow rates and electrometer sensitivity so that
4 µL cocaine standard solution gives 40–60% full scale deflection.
Retention time of cocaine relative to internal standard is ca 0.65.
(b) Dibasic potassium phosphate.—10%. Dissolve 5 g K2HPO4
in 50 mL H2O, and mix well.
(c) Dilute hydrochloric acid.—0.1M. Dilute 7.0 mL HCl to 1 L
with H2O, and mix well.
(d) Internal standard solution.—0.8 mg/mL. Dissolve 80 mg
tetracosane in 100 mL CHCl3, and mix well. (USP Cat. No. 143008
or Sigma-Aldrich Cat. No. C5776.)
(e) Co caine hy dro chlo ride stan dard so lu tion.—1 mg/mL.
Accurately weigh ca 10 mg cocaine⋅HCl into 10 mL volumetric
flask, and dilute to volume with internal standard solution. Prepare
fresh every 3 months. Store in refrigerator.
C.
Plug column exit with silanized glass wool. Apply vacuum to exit
end, and slowly add packing material through column inlet while
gently tapping column. Fill to within 1 cm of column inlet, and plug
with silanized glass wool. Condition column overnight at 260°C with
slow stream of N2. Saturate column with cocaine by making
successive 4 µL injections of cocaine standard solution until cocaine:
internal standard ratio differs by <3% from preceding standard
injection.
D. Determination
Accurately weigh ca 250 mg finely ground test portion and dilute
quantitatively with 0.1M HCl to estimated cocaine⋅HCl concentration
of 1 mg/mL. Mix well and let any insoluble material settle. Pipet 2 mL
test solution, 2 mL internal standard solution, and 1 mL 10% K2HPO4
into test tube; stopper, and shake vigorously. Let layers separate. Pipet
2 mL cocaine standard solution, 2 mL 0.1M HCl, and 1 mL 10%
K2HPO4 solution into an other test tube, stopper, and shake
vigorously. Let layers separate. Inject 4 µL CHCl3 (bottom) layer of
test and standard solutions, each in duplicate, using 10 µL syringe.
Cocaine⋅HCl, % = (P/P ′) × (B′/B) × (C/W) × D × 100
where P and P ′ = average areas or peak heights of analyte and
standard cocaine⋅HCl, respectively; B and B′ = average areas or
peak heights of internal standard in test and standard solution,
respectively; C = mg cocaine⋅HCl/mL in standard solution; W = mg
test material; and D = dilution factor for test material.
Cocaine⋅HCl/tablet, mg = cocaine⋅HCl, %
× average tablet weight in mg/100
Reference: JAOAC 61, 473, 683(1978).
CAS-53-21-4 (cocaine hydrochloride)
 2005 AOAC INTERNATIONAL