Detection of the Adulteration of Traditional Alcoholic Beverages by the Separation and Determination of Alprazolam, Chloralhydrate and Diazepam Using Reversed-Phase High-Performance Liquid Chromatography

Abstract

Food is essential not only for the sustenance of life, but also for the maintenance of good health. However, contaminated or adulterated food is a major source of human illness, and results in a serious loss of nutrition. 1 Thus, the quality and safety of food assumes great significance in the promotion of optimum health. Toddy is one of the alcoholic drinks traditionally prepared by the fermentation of sap or exudate collected by slicing off the tip of unopened flowers of either a coconut or a palm tree. 2 It is a social and local drink produced and consumed throughout Asia, particularly India, Sri Lanka and Bangladesh. Due to the increase in consumer demand and the shortage of coconut or palm tree, it has often been diluted and adulterated with chemical substances, such as chloral hydrate (CHL), diazepam (DIA) or alprazolam (ALP) belonging to nonbarbiturates and benzodiazepines, respectively. 3,4 These drugs are generally considered to be dangerous when consumed along with alcohol, and have proved to be fatal poisonings according to the recent analysis of forensic toxicological investigations conducted by Koski et al. 5 Further, it was reported that female drinkers are at increased risk to abuse benzodiazepines when compared to men. 6 A case of alprazolam poisoning present with coma and respiratory depression in a young girl was also reported. 7 These psycotropic substances have a sedative action and are added illicitly to toddy to increase its potency. The adulterated toddy is prohibited for consumption under the laws of excise, which necessitates the development of suitable analytical methods for the simultaneous detection and determination of the levels of psycotropic substances in fermented food and alcoholic beverages.Fujiwara test, which gives an intense red color by the action of pyridine in the presence of an alkali, is generally used to identify CHL in toddy. 8-10 A number of assays have shown CHL to be genotoxic 11 and hepato-carcinogenic in male mice. 12Due to its widespread potential for human exposure, US FDA has declared CHL as a priority chemical for an in-depth toxicological evaluation. As a part of this program, a capillary gas chromatography with electron-capture detection was developed for the determination of CHL and its metabolites in blood plasma of mice and rats. 13 Two liquid-chromatographic methods based on reversed-phase and anion-exchange mechanisms were also reported in the literature. 14 However, both of the methods are preceeded by the derivatization of CHL with dansylhydrazine and o-(4-nitrobenzyl)hydroxylamine, followed by detection using UV and suppressed conductivity detectors. A spot test using Dragendorffs reagent containing bismuth nitrate and potassium iodide in an acetic acid medium produces an orange-red color, indicating the presence of DIA. 15The colorimetric determination of DIA in pharmaceutical preparations was described by Baggi et al. 16 However, these tests are not specific because similar colors are produced by a number of organic compounds containing halogens...

A rapid CZE method for the analysis of benzodiazepines in spiked beverages

“…While the CZE-DAD method was capable of quantifying DIA, NIT, FLU, TEM and OXA, it lacked the sensitivity required to detect ALP at a concentration designed to simulate a standard drink spiked with a prescription tablet of this particular drug. Recoveries of 79.4-135.8% were obtained for [13,39,40], the PDDAC/DScoated method offers considerably shorter run times. Another advantage of this method is that it does not require an extraction step.…”

“…A similar method was employed by Chen and Hu [39] who extracted benzodiazepines from several beverages using chloroform or dichloromethane prior to analysis by direct electrospray probe/MS. Rao et al [40] employed an HPLC method for the determination of ALP, DIA and chloral hydrate in the alcoholic beverage toddy. The analysis time was 15 min and the only sample pretreatment required was filtration through 0.45 mm Nylon membranes.…”

Automated determination of diazepam in spiked alcoholic beverages associated with drug-facilitated crimes

“…This reference methodology was the one recommended by the United States Pharmacopoeia (USP) [13] for the determination of diazepam pure drug. However, this procedure was modified in conformity with the work of Rao et al [9] due to the scarce worldwide availability of acetonitrile.…”

“…In literature several published methods for analysis of benzodiazepines in beverages can be found based either on Gas Chromatography/Mass Spectrometry (GC-MS) [6], Capillary Zone Electrophoresis/Diode Array (CZE-DAD) [4], Direct Electrospray Probe/Mass Spectrometry (DEP/MS) [7], High Performance Liquid Chromatography/Diode Array (HPLC-DAD) [8,9], High Performance Thin-Layer Chromatography (HPTLC) [10], etc. The methods above mentioned are very precise and present high sensitivity, however they are only performed in very special laboratory conditions, and they are laborious and require skilled operators.…”

Novel reductive–reductive mode electrochemical detection of Rohypnol following liquid chromatography and its determination in coffee

“…A number of methods have been developed, such as immunoassay [9], GC/MS [10] and HPLC [11,12]. However, there are relatively few reports on the determination of benzodiazepines in beverage samples [13][14][15][16][17][18]. One powerful technique, which has received little attention for the determination of benzodiazepines, is high performance liquid chromatography with dual electrode detection (LC-DED).…”