Hi Folks,

I stumbled across the following, which to me is a real eye-opener:

http://www.sciencedirect.com/science?_o ... fd12c642a3

In particular, this statement caught my attention:

"It was found that naturally occurring GHB and GBL were detected in those beverages involving the fermentation of white and particularly red grapes. No GHB or GBL was detected in other drinks such as beer, juice, spirits or liqueurs". Now I don't think either GHB or GBL have been mentioned before on this forum; I could be wrong. However, Dr Olivier Ameisen (of baclofen fame) produced the following hypothesis:

http://alcalc.oxfordjournals.org/cgi/re ... m058v1.pdf

So, putting these pieces of the jigsaw together, there may be a specific reason why I prefer wines over other alcoholic drinks. I should add that I don't drink much red wine as they tend to be 'heavier' than white wines with the result that I am more likely to get hangovers from red wines. Port is particularly bad ( ) in this respect.

V.

I don't know V, it sounds reasonable.

Here's the full study:


The presence of gamma-hydroxybutyric acid (GHB) and gamma-butyrolactone (GBL) in alcoholic and non-alcoholic beverages

Simon ElliottCorresponding Author Contact Information, E-mail The Corresponding Author and Victoria Burgess

Regional Laboratory for Toxicology, Sandwell and West Birmingham Hospitals NHS Trust, City Hospital, Dudley Road, Birmingham B187QH, UK

Received 27 January 2005;
revised 18 February 2005;
accepted 28 February 2005.
Available online 19 March 2005.

Abstract

Gamma-hydroxybutyric acid (GHB) and its precursor gamma-butyrolactone (GBL) are regularly implicated in instances of surreptitious drug administration, particularly in beverages (so-called “spiked drinks”). In order to assist in the interpretation of cases where analysis of the actual beverage is required, over 50 beverages purchased in the UK were analysed for the presence of GHB and GBL. It was found that naturally occurring GHB and GBL were detected in those beverages involving the fermentation of white and particularly red grapes. No GHB or GBL was detected in other drinks such as beer, juice, spirits or liqueurs. GHB/GBL was detected in red wine vermouth (8.2 mg/L), sherry (9.7 mg/L), port (GBL), red wine (4.1–21.4 mg/L) and white wine (<3–9.6 mg/L). The presence of GHB/GBL did not appear to be influenced by the alcohol content or the pH of the beverage. In addition, the concentration in wines did not appear to be related to the geographical origin of the grape type. This is believed to be the first published data concerning the endogenous presence of GHB and GBL in the beverages described.

Keywords: Gamma-hydroxybutyric acid; Gamma-butyrolactone; Beverages
Article Outline

1. Introduction
2. Materials and methods

2.1. Chromatographic methods
2.2. Beverage analysis

3. Results and discussion
Acknowledgements
References

1. Introduction

Gamma-hydroxybutyric acid (GHB), also known as “Liquid Ecstasy” has been implicated in many cases of suspected surreptitious administration, potentially for the purpose of increasing victim vulnerability to sexual assault [1]. Due to the rapid dissolving properties of GHB powder resulting in a colourless solution when in water, the perceived route of administration is typically associated with the adulteration of a victim's beverage. The precursor solvent gamma-butyrolactone (GBL) may be used as an alternative as it is rapidly converted to GHB in the body [2]. The use of these compounds in this way can be a very dangerous practice as the exact concentration of the resultant solution is largely unknown and may result in serious and potentially fatal GHB intoxication. This would particularly be the case if the “spiked” beverage was alcoholic or if the victim was/had been drinking alcohol prior to the incident. Following an accusation of suspected “spiked drink”, as part of the investigation and if available, the actual beverage may be analysed to determine the presence of drugs (e.g. GHB). The necessary dose of GHB to produce incapacitation or even disinhibition (approximately >2 g) would be associated with high concentrations in beverages due to the relatively small liquid volume of most drinks. Nonetheless, it would be necessary to be aware of any naturally occurring GHB or GBL in the beverage. GHB is believed to be present in ripe fruits and meat and Collison et al. recently presented data from the United States for various beverages and foodstuffs [3] and [4]. It was found that GHB was present at its highest concentration in red (2.00–23.00 mg/L) and white wine (0.65–9.53 mg/L), followed by vinegars (0.83–11.25 mg/L), soy sauce (2.79–18.10 mg/L), liqueurs (<0.25–4.20 mg/L), various non-distilled drinks (1.88–6.68 mg/L), beer (<0.25–2.10 mg/L) and coffee (0.52–2.09 mg/L). No GHB was detected (<0.50 mg/L) in any of the distilled alcoholic drinks (e.g. whisky) or juices tested (except grape juice). There are currently no available or published data regarding specific concentrations in alcoholic and non-alcoholic beverages commercially available in the UK.

Another aspect of the necessity of information relating to the natural presence of GHB in drinks involves the increasing availability of alleged detection kits for drugs (including GHB) in beverages. These products are marketed for individuals to be able to “test” their drink to determine whether a drug has been added. Apart from the immediate flaws in the process (i.e. only detect a limited number of compounds), additional problems include: (i) it may be “negative” at the time of testing but may require continual testing, (ii) the chemistry involved in detection may lead to false positive/negative results and (iii) the lack of any immediate confirmation [5] and [6]. It is also conceivable that depending on the nature or sensitivity of the assay, the presence of certain residual/endogenous compounds may also lead to false positive findings. The possibility of false positive or negative results could have important medical, social and legal ramifications. Therefore, there is both a forensic and public requirement for data concerning the presence of compounds, in particular GHB, in alcoholic and non-alcoholic beverages. This short communication presents novel data from a study of over 50 beverages purchased within the UK.
2. Materials and methods

Beverages were obtained from various licensed premises and stores. In some cases multiple products/brands were analysed of the same drink group (e.g. lager). A 5 mL aliquot of each beverage was retained and stored at 4 °C prior to analysis (typically analysed within 3 days following collection). Samples were initially analysed using gas chromatography with flame ionization detection (GC–FID). Gas chromatography with mass spectrometry (GC–MS) was used for confirmation of “positive” GC–FID samples in addition to some randomly selected beverages.
2.1. Chromatographic methods

GC–FID and GC–MS analyses were performed using previously published methods [7] and [8]. In summary; GC–FID detected the presence of GHB and GBL as “total GBL” as it involved complete conversion to GBL prior analysis. This involved acidification with 6 M H2SO4 and extraction with chloroform, including 100 mg/L hexanoic acid as an internal standard. GC–MS detected the presence of GHB following trimethylsilane (TMS) derivatisation and analysis in the full scan mode. GHB-D6 was included as an internal standard (in 0.05 M H2SO4) followed by extraction with acetonitrile.
2.2. Beverage analysis

GHB calibration standards of 1, 2.5, 5, 10 mg/L were prepared in blank equine plasma (pre-screened <1 mg/L GHB) along with an internal quality control standard of 7.5 mg/L. Plasma presented less derivatisation problems for GC–MS compared to the use of aqueous standards (e.g. water) and had previously been shown to be valid for the analysis of more aqueous specimens (e.g. urine) [9].

Beverages were directly extracted for GC–FID analysis; however, they required three-fold dilution with blank equine plasma prior to GC–MS analysis. This was due to observed chromatographic and mass spectral interferences/extraneous peaks following TMS derivatisation of undiluted/non-matrix matched samples (in particular, wine). Such interferences did not occur when using plasma. Consequently, this dilution factor reduced the sensitivity of the GC–MS assay from 1 to 3 mg/L.

The pH of the red and white wines were measured using a Jenway pH Meter 3310 (Dunmow, Essex, UK).
3. Results and discussion

Table 1 shows GHB/GBL was detected in only a few of the non-alcoholic and alcoholic beverages analysed in this study, primarily those involving grapes and fermentation processes in their production such as wine (GHB <3–21 mg/L), port (GBL) and sherry (GHB 9.7 mg/L). No GHB (<3 mg/L) or GBL (<5 mg/L) was detected in beer, distilled alcoholic drinks or fruit juices. The highest GHB concentrations were found to be in red wine, ranging from 4.1 to 21.4 mg/L (mean = 12.6 mg/L). This is compared to white wine which had an associated GHB range of <3–9.6 mg/L (mean = 5.7 mg/L). For each of the 26 red and white wines analysed, their country of origin, alcohol content and pH was noted (Table 2). There did not appear to be any obvious correlation between these factors and the concentration of GHB found. In theory, a very acidic pH would shift the chemical equilibrium towards GBL, thus lowering the GHB concentration. Although this may be a reason for the reduced GHB concentrations in white wine, the overall similarity of pH values between the red and white wines reduces the likelihood of this explanation. The highest concentration of GHB was detected in a red wine from Chile (21.4 mg/L) and the lowest was from a white wine from France (not detected <3 mg/L). With the additional detection of GHB/GBL in red wine based vermouth and port, it appears red grapes (or the particular production processes associated with them) contain more GHB/GBL or result in the production of more GHB/GBL during the fermentation process than “white” grapes. There was an associated absence of GHB/GBL (<5 mg/L) in both white and red grape juice but this does not necessarily provide any additional evidence for the provenance of the GHB.
Table 1.

Presence of GHB/GBL in various beverages
Beverage Alcohol content (%, v/v) GHB concentration (mg/L) GBL detected? (LOD = 5 mg/L)
Aftershock blue 40 ND No
Amaretto 28 ND No
Bitter 4 ND No
Cream liqueur 17 ND No
Bourbon 40 ND No
Brandy 40 ND No
Cider 5.5 ND No
Drambuie 40 ND No
Gin 40 ND No
Red wine vermouth 17 8.2 Yes
Dark rum 40 ND No
White rum 37.5 ND No
Sherry 17.5 9.7 Yes
Vodka 37.5 ND No
Vodka + juice 5 ND No
Vodka schnapps 4.4 ND No
Whisky 40 ND No
Cranberry juice None ND No
Grapefruit juice None ND No
Orange juice None ND No
Pineapple juice None ND No
Tomato juice None ND No
Tonic water None ND No
Tequila 38 ND No
Absinthe 70 ND No
Lager 4 ND No
Port 20 ND Yes
Red grape juice None ND No
White grape juice None ND No
Red wine 12–14 4.1–21.4 (mean 12.6) Yes
White wine 9.5–13.5 <3–9.6 (mean 5.7) Yes
Full-size table

LOD: limit of detection; ND: not detected (GHB limit of detection = 3 mg/L).

View Within Article



Table 2.

GHB concentrations in red and white wine
Beverage Country of origin Alcohol content (%, v/v) pH GHB concentration (mg/L) GBL detected? (>5 mg/L)
Red wine 1 Chile 14 3.70 21.4 Yes
Red wine 2 Australia 14 3.37 10.6 Yes
Red wine 3 Argentina 13 3.43 19.2 Yes
Red wine 4 Australia 14 3.47 15.0 Yes
Red wine 5 Romania 12 3.53 13.5 Yes
Red wine 6 USA 13.5 3.59 15.8 Yes
Red wine 7 USA 13.5 3.24 13.7 Yes
Red wine 8 USA 13.5 3.49 14.9 Yes
Red wine 9 Spain 13.5 3.68 10.2 Yes
Red wine 10 Spain 13 3.58 11.1 Yes
Red wine 11 Spain 12.5 3.46 9.6 Yes
Red wine 12 France 12 3.52 12.1 Yes
Red wine 13 Spain 12.5 3.49 4.1 Yes
Red wine 14 Italy 13 3.25 10.2 Yes
Red wine 15 Argentina 14 3.51 10.8 Yes
Red wine 16 Italy 13 3.47 9.6 Yes

White wine 1 New Zealand 10.5 3.26 8.4 Yes
White wine 2 Australia 12 3.00 5.9 Yes
White wine 3 South Africa 13.5 3.47 8.5 Yes
White wine 4 South Africa 3.51 6.8 Yes
White wine 5 South Africa 12.5 3.51 3.5 Yes
White wine 6 Germany 9.5 3.41 9.6 Yes
White wine 7 Australia 12.5 3.30 5.1 Yes
White wine 8 France 11.5 3.17 5.7 Yes
White wine 9 France 12 3.29 <3 Yes
White wine 10 Italy 12 3.19 3.3 Yes
Full-size table

View Within Article


With regard the implications of these results, the data indicate there may be interpretative issues when investigating suspected surreptitious GHB administration via the adulteration of wine. Whereby it is likely that naturally occurring GHB will be detected. Due to the observed variation of concentrations even within grape types it is recommended that specific analyses be performed of purchased products of the same type as that implicated in the case. Other beverages do not seem to contain any endogenous GHB and therefore detection (e.g. at concentrations above 10 mg/L) may indicate potential exogenous introduction to the drink. As previously mentioned, in order for an effective dose to be administered it is conceivable that a far higher concentration would be present in the drink and should therefore be easily distinguishable from any naturally occurring GHB. For example, 2 g of GHB in 250 mL of liquid, a large glass of wine, would result in a beverage concentration of 4000 mg/L. Even 0.5 g of GHB in 250 mL would produce a concentration of 1000 mg/L. In the UK, for one pint of beer (not, vert, similar568 mL) 0.5 g of GHB would produce a concentration of not, vert, similar880 mg/L. However, if attempts have been made by the perpetrator to remove any added GHB (e.g. by rinsing the container), there is still the possibility that low concentrations of residual GHB may be found.

In the case of commercially available drug testing kits for drinks, there are very limited data concerning the limit of detection for any of the drugs purported to be detected, especially GHB. During a study of the capabilities of the enzymic detection of GHB (utilizing GHB dehydrogenase) Bravo et al. indicated a maximal detection limit of 0.037 mg GHB/mL (37 mg/L) [6]. Therefore, under experimental conditions and using purified enzyme, none of the drinks analysed in this study would necessarily provide a “positive” result. However, as the exact chemistry, cross-reactivity and sensitivity of “drink testing kits” is largely unknown and unpublished, the possibility remains that particularly in the case of wines, a “false positive” result may be achieved due to the presence of naturally occurring GHB in the drink itself. Further evaluation of such kits should be performed either by the manufacturers or other researchers in order to better interpret any findings and determine the usefulness of these products.
Acknowledgments

The authors would like to thank Mr. Christopher Smith and Mrs. Jody Griffiths for their assistance during the study.


References available upon request -E

Hi Lo0p,

Thanks a million!

I was hoping you may have access to the full article. I only had to wait for a couple of hours and, hey presto, here it is. Out of interest, if I PM my e-mail address to you, would you mind e-mailing me the full article?

Once again, thank you.

V.

Thanks Lo0p.
This may explain my almost exclusive preference for Red wine over all other alcoholic drinks.
However here I am in week 21 of TSM and my preference is changing to White Wine.
As we here in Oz are coming into Summer, it may have something to do with the weather.
Warm Red wine is not as palatable as a chilled White.

Extinction Continues

Sure V. It's a lot easier to read in the PDF format too. -E

Interesting.

Because of the additional chemicals, I wonder how many of the Cured are wine drinkers versus another DOC. Personally, I never liked wine, even when I was an alchy. There was always wine in the house, but I refused to drink it. Anyway, I've successfully gone through 9 months of TSM entirely on one beverage, beer. My addiction is gone, thus I am technically cured. Do the additional chemicals in wine perhaps add another parameter that may affect TSM?

So, how many other cured ones are mainly or exclusively wine drinkers?

Bob

Could someone please paraphrase the article? It is so technical and full of scientific jargon, my eyes just crossed and glazed over after a couple of paragraphs...similar to reading a how-to manual on computers...:-/

Hi Folks,

When I started this thread, I was hoping that it would generate interest and it certainly seems to be doing that. Although I entitled the thread "Is this why I'm a wine-drinker?", I was hoping to find out what others' drink of choice is/was. Lo0p, would it be possible to add a column to your spreadsheet in order that members can provide this information should they wish to do so?

So far, I see we've heard from two members that are on the cured list. Bob is a beer drinker whilst AJ is primarily a wine drinker. It would be good to hear from others.

V.

Sure I'll add a DOC column to the spreadsheet. Did you still want me to email you that PDF?