Phenylacetone is an organic compound with the chemical formula C6H5CH2COCH3. It is a colorless oil that is soluble in organic solvents. This substance is used in the manufacture of methamphetamine and amphetamine, where it is commonly known as P2P. BMK.
What is Phenylacetone?
Phenylacetone is an organic compound with the chemical formula C₆H₅CH₂COCH₃. It is also called BMK. This substance is used or precursor in the manufacture of methamphetamine and amphetamine, where it is commonly known as P2P. The cas number is CAS NO.:103-79-7.
Major Uses of Phenylacetone
1-Phenyl-2-propanone (P-2-P), also known as Phenylacetone or benzyl methyl ketone (BMK), is a colorless or slightly yellowish liquid. It presents a density similar to that of water as well as a pleasant scent. Even if there are few legitimate uses of BMK or Phenylacetone such as in the production of the pharmaceutical drug propyl-hexedrine, most frequently BMK or Phenylacetone is used as an illicit compound. Actually, BMK is identified by classical methods such as gas chromatography, NMR or HPLC. These methods are costly, time-consuming and require the presence of trained operators. It appears obvious that there is an urgent need to develop a new easy and fast method that allows us to detect the presence of traces of BMK or Phenylacetone. In this work, a new chemically synthesized BMK derivative covalently attached to an immunological carrier was used for producing antibodies against the BMK or Phenylacetone molecules. A fluorescence polarization-based bioassay was developed by using the produced anti-BMK antibodies and the BMK or Phenylacetone derivative. The assay exhibits interesting analytical performances with a limit of detection of less than 100 nM and an almost linear response up to 600 nM. Interestingly, the proposed assay could be performed using a customizable portable instrumentation and could be used by non-instructed personnel at custom borders and checkpoints or for quick spot-checks.
Materials and methods of Phenylacetone
All reagents were of the highest commercially available qualityand used as received. 1-[3-(Dimethylamino)-propyl]-3-ethyl-carbodiimide (EDC), bovine serum albumin (BSA; fraction V),carboxymethoxylamine hemihydrochloride, benzyl methylketone (BMK), EAH Sepharose 4B resin and buffers werepurchased from Sigma-Aldrich. The ﬂuorescent probe CF488Awas purchased from Biotium Inc. Nitrocellulose transfermembrane Protran from Schleicher & Schuell and ECL detectionreagents from Amersham Biosciences were used in Western blotexperiments. Goat polyclonal to rabbit IgG–HRP conjugate(secondary antibody) was from Abcam. The NMR spectra of theBMK derivative were recorded on a Varian Gemini 200 (200 MHz).
Synthesis of (1-methyl-2-phenyl ethylideneaminooxy)acetic acid
Benzyl methyl ketone (99% Sigma Aldrich, 612 mL, 4.56 mmol)was reﬂuxed with ten equivalents of o-carboxymethoxylaminehemihydrochloride (99% Sigma Aldrich, 5.00 g, 45.7 mmol) in6 mL of pyridine–H2O–methanol (1 : 1 : 4) for 2 h. The reactionwas followed by thin layer chromatography (n-hexane–ethylacetate 8 : 2 + HCOH drops). Pyridine and other solvents wereremoved by toluene co-distillation by a rotavapor. The productof the reaction was re-suspended in 5 mL of H2O at pH 9.00basiﬁed with NaOH and washed with 3 10 mL of dichloro-methane. Then the aqueous phase was acidiﬁed with HCl to pH3.0 and the product extracted in 3 15 mL of dichloromethane.The solvent was dried over NaSO4and vacuum distilled to leavean oily product. In order to remove reaction by-products and theexcess of carboxymethoxylamine, ﬂash chromatography wasperformed on a silica column (silica gel 60 M 0.04–0.0063 mm)with eluent CH2Cl2–CH3OH 9 : 1 + HCOOH drops (isolatedyield 45%).1H NMR (400 MHz, CDCl3): d 1.82 ppm (s, 3H), d3.48 ppm (s, 1H), d 3.76–3.78 ppm (m, 1H), d 4.67 ppm (s, 2H), d7.40–7.70 ppm (m, 5H).
Synthesis of BSA conjugate (BMK–BSA)
To avoid interference by the carrier protein in the polyclonalantibody detection process, BMK-oxime was conjugated to theserum albumin (BSA). The following procedure was used: 0.484mmol BMK-oxime was dissolved in ethanol and mixed with anaqueous solution of 4.84 mmol EDC. The solution was diluted upto 840 mL with 10 mM phosphate buffer at pH 6 and incubatedfor 20 min at room temperature. Finally the solution was incu-bated with 4.84 nM BSA dissolved in 160 mL of 10 mM phos-phate buffer at pH 6.
The reaction mixture was incubated atroom temperature under continuous stirring and then dialyzedagainst 0.5 L of 10 mM phosphate buffer at pH 7.4 for 4 dayswith daily buffer changes. Antibody production and IgG puriﬁcationA rabbit was immunized following a standard protocol by intra-dermal injection. After the immunization period, the rabbit wassacriﬁced and its blood collected and centrifuged to separateblood cells from serum. A 2.0 mL sample of serum of the rabbitwas diluted 1 : 1 in 50 mM Tris–HCl at pH 7.0 (binding buffer) and applied to 0.5 mL of resin protein A SepharoseTM 4 FastFlow (GE Healthcare).
The IgG fraction was puriﬁed accordingto the manufacturer’s instructions. The IgG fraction was elutedwith glycine (0.1 M) at pH 2.8 and immediately buffered with1.0 M Tris–HCl at pH 8.8. Elution of IgG proteins was moni-tored by absorbance at l¼278 nm and SDS PAGE was carriedout to evaluate the purity of the samples (data not shown). Phenylacetone
Precursors for amphetamine and methamphetamine
Phenylacetone and its pre-precursors
Seizures of Phenylacetone or BMK decreased steadily. In response to strict controls preventing the diversion of BMK or Phenylacetone, resourceful producers have, for a number of years, been making Phenylacetone or BMK from other substances — the so-called ‘pre-precursors’. Despite these developments, there are recent indications that BMK or Phenylacetone may be re-emerging as a significant precursor in its own right. Phenylacetone
China reported the seizure of almost 5 500 litres of BMK or Phenylacetone in 2013, despite it having been absent from the market for some years previously, and it is believed that the BMK seized was destined for Spain (INCB, 2015a). Phenylacetone
In addition, at the end of March 2015, Polish law enforcement authorities discovered 7 000 litres of BMK in Warsaw which had been shipped from China to Poland via Germany, with the declared destination being a company in the Czech Republic. In the subsequent investigation, links with an amphetamine production site in the Netherlands were established (Europol, 2015e). Phenylacetone
Phenylacetone (BMK) can be made from other chemicals, e.g. from phenylacetic acid (PAA) and either acetic acid or acetic anhydride, and by the so-called ‘nitropropene route’, using benzaldehyde and nitroethane (Krawczyk, 2005); and in the past large quantities of a white powder known as ‘BMK or Phenylacetone bisulphite adduct’, sourced from Russia, have been seized (Europol, 2009). Phenylacetone
One of the most important developments in the story of BMK was the discovery that it can be produced from alpha-phenylacetoacetonitrile (APAAN), an innovation that was probably driven by erratic availability of other precursors. Phenylacetone
Since this development was first identified in 2009, APAAN has become an increasingly important pre-precursor and is imported into Europe primarily from China (on some occasions misdeclared as other products or substances) and converted into BMK using large quantities of acids, often in dedicated conversion laboratories. Phenylacetone
In 2013, large seizures of APAAN were made in Europe, notably 36 tonnes in the Netherlands and 5.4 tonnes in Belgium. In Poland, a laboratory for the conversion of APAAN to BMK or Phenylacetone was found and 1.4 tonnes of APAAN was seized (INCB, 2015a). In December 2013, APAAN was scheduled as a precursor under European legislation, and international control followed in October 2014. Phenylacetone
Despite these control measures, APAAN continues to be seized; in March 2014, Bulgarian authorities seized almost 1 tonne of APAAN that had arrived on a truck from Turkey, misdeclared as ‘soluble dyes’ (INCB, 2015a). Seizure data suggest that large quantities of APAAN were stockpiled in Europe prior to controls being introduced and, consequently, BMK or Phenylacetone made from APAAN continues to be used by OCGs for the large-scale production of amphetamines (see Figure 6.7). Contact us for more details about Phenylacetone.