Imprinted polymeric NANO materials for the purification and determination of methamphetamine or amphetamine and synthesis method thereof

Abstract

The invention relates to a polymeric nanomaterial that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale, as well as to the synthesis method thereof.

Description

[0001] DESCRIPTION

[0002] IMPRINTED POLYMERIC NANO MATERIALS FOR THE PURIFICATION AND DETERMINATION OF METHAMPHETAMINE OR AMPHETAMINE AND SYNTHESIS METHOD THEREOF

[0003] Technical Field of the Invention

[0004] The invention relates to a polymeric nanomaterial that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale, as well as to the synthesis method thereof. Thus, the invention enables the chromatographic purification of MeA or AMP in licensed production processes to be performed in a single step, while also providing a nanomaterial that can be reused. In addition, due to its high affinity for MeA or AMP, the polymeric nanomaterial subject to the invention can be used as a selective material in commercial diagnostic and biosensor systems.

[0005] State of the Art

[0006] Narcotics is a general term for drugs with psychoactive effects. With the abuse of its psychoactive effect, the term narcotics takes on a different dimension [1], A narcotic drug is defined as a toxic substance which, when used, causes changes in sensation and behavior in the central nervous system; causes addiction in advanced stages; causes mental and physical reactions in its absence; causes economic and social collapse in the individual and society; and whose use, possession and sale are prohibited by law. Narcotics that stimulate the central nervous system are examined in general, they are divided into three groups: natural stimulants, synthetic stimulants, and hallucinogens. Examples of synthetic stimulant narcotic drugs include stimulants such as amphetamine, methamphetamine, and ecstasy. Synthetic narcotics are synthesized in the laboratory from a chemical starting material without any dependence on plants or natural processes.

[0007] Amphetamines, one of the oldest known synthetic psychoactive substances, are commonly referred to as "speed" and are widely used as recreational drugs [2], Amphetamine-type stimulants are substances such as amphetamine, methamphetamine, metheydrine and "ecstasy" (3,4-methylenedioxymethamphetamine(MDMA) and its analogues), which are internationally controlled under the 1971 Convention on Psychotropic Substances [3]. However, Methamphetamine (MeA) and Amphetamine (AMP) are studied under license for medical, scientific, and forensic purposes as well as for illegal use. Licensed studies of these analytes take place mainly in areas such as drug development, forensic analysis, toxicology, and clinical research. . In such a case, licensed production of MeA and AMP is also carried out by licensed companies (pharmaceutical and chemical companies). Since the MA and AMP produced are obtained as a result of organic synthesis, an enrichment or chromatographic purification (HPLC, GC) process is applied to purify the products from impurities. However, HPLC and GC analyses are often time-consuming, especially in complex matrices the separation of impurities takes time. During purification, part of the target product may be lost. For example, yields are reduced due to retention in columns or loss with solvent. Furthermore, the same sample needs to be analyzed more than once to confirm the accuracy of the results obtained. These techniques are not economically and operationally viable when large quantities need to be purified at the industrial level.

[0008] Polymeric nanomaterials offer many advantages as column fillers used in chromatographic techniques in analytical chemistry. These advantages arise from the physical, chemical, and surface properties of the materials and enable higher efficiency and precision in analytical processes. Polymeric nanomaterials can recognize and separate specific target molecules by molecular imprinting. Surfaces modified with functional groups make strong and selective bindings with certain types of molecules. The high surface area of polymeric nanomaterials increases the interaction with target molecules by providing more active sites and these materials are reusable and stable in long-term studies. Nanopolymers with high specificity (affinity) for target molecules such as methamphetamine (MeA) and Amphetamine (AMP) are used in different technical fields such as chemistry, biotechnology, and forensic sciences. Molecularly imprinted polymers are designed to have specific binding sites with the target molecule (e.g. MeA or AMP). MIPs are artificial receptors in which the target molecule is used as a "template" and produced by a polymerization process together with cross-linking monomers. These polymers can bind to MeA and AMP with high specificity as they contain specialized cavities at the molecular size.CN102016814A patent application in the state of the art relates to the preparation of micron and / or nanoscale particles. It is noted that the functionalized particles disclosed in the present invention comprise substantially similar spatial and chemical properties of molecularly imprinted templates. It is disclosed that the adherent layer may contain 2-hydroxyethyl methacrylate and that a template selected from enzymes, proteins, antibiotics, antigens, nucleotide sequences, amino acids, drugs, biological agents, nucleic acids and combinations thereof will be used for molecular imprinting. Said drugs include amphetamine and methamphetamine.

[0009] Another patent application numbered KR102437215B1 in the state of the art discloses a molecularly imprinted polymer (MIP) based electroanalytical detection platform comprising a chalcogenide-loaded cobalt metalorganic framework and a metal conducting polymer nanocomposite, and a use thereof. Said document uses 2-hydroxyethyl methacrylate as a functional monomer. The sample applicable to the molecularly imprinted polymer-based detection platform is said to be amphetamine.

[0010] Due to reasons such as limitations and shortcomings of the solutions in the current technique, the purification of industrially licensed MeA and AMP at high purities using HPLC and GC methods, which are time-consuming and costly processes, requiring complex equipment infrastructure and specialized personnel, demanding high accuracy and precision, and necessitating sustainability of the processes and the ability to pass small sample quantities through the columns, a development in the relevant technical field has become necessary.

[0011] Summary and Objects of the Invention

[0012] The invention describes a polymeric nanomaterial that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale as well as to the synthesis method thereof. Thus, the invention enables the chromatographic purification of MeA or AMP in licensed production processes to be performed in a single step, while also providing a nanomaterial that can be reused. In addition, due to its high affinity for MeA or AMP, the polymeric nanomaterial subject to the invention can be used as a selective material in commercial diagnostic and biosensor systems.The object of the invention is to provide polymeric nanomaterials with high affinity that can be used as column fillers. HPLC and GC methods are actively used to obtain high purities of MeA and AMP produced under industrial license. With HPLC and GC methods, low amounts of samples can be purified with long-lasting processes. The purification process with the materials subject to the invention is fast and affordable.

[0013] An object of the invention is to provide a nanomaterial with high chromatographic separation power, binding efficiency, and selectivity, biocompatibility, low cost, and a strong skeletal system (polymer chain). In the invention, nonatechnology is added to MIP technology to obtain a more advanced material and a material with a high surface area. In the pre-complex step, the first step in MIP technology, high binding efficiency (weak secondary interactions) is achieved between MeA and AMP with NMT designated as the functional monomer. The HEM monomer selected as the main monomer gives a nano-structure, high binding efficiency, high selectivity, biocompatibility, low cost, and a strong backbone system (polymer chain) to our polymer. Thus, the separation power (chromatographic separation power) of the polymeric nanomaterial is maximized.

[0014] Description of the Drawings

[0015] Fig. 1. FTIR measurements of polymeric nanoparticles A-1, A-2, and A-3, (respectively 25 pL, 50 pL, 80 pL NMT functional monomer-containing AMP-imp-p(HEM-NMT) and M-1, M-2, and M-3 (respectively 25 pL, 50 pL, 80 pL NMT functional monomer-containing MeA-imp-p(HEM-NMT).

[0016] Fig. 2. Zeta dimension and potential analysis.

[0017] Fig. 3. Zeta dimension and potential analysis.

[0018] Fig. 4. SEM analysis results for MeA-imp-p(HEM-NMT), AMP-imp-p(HEM-NMT), and NIP.

[0019] Fig. 5. AFM (Atomic mass microscopy) results A) AMP-imp-p(HEM-NMT) B) MeA-imp-p(HEM-NMT).

[0020] Fig. 6. Adsorption studies of synthesized polymeric nanoparticles: A) AMP-imp_p(HEM-NMT), B) MeA-imp-p(HEM-NMT)) adsorption graph

[0021] Fig. 7. Cross-selectivity study for AMP and MA for MeA-imp-p(HEM-NMT) and AMP-imp-p(HEM-NMT).Detailed Description of the Invention

[0022] The invention relates to a polymeric nanomaterial that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale, as well as to the synthesis method thereof. Thus, the invention enables the chromatographic purification of MeA or AMP in licensed production processes to be performed in a single step, while also providing a nanomaterial that can be reused. In addition, due to its high affinity for MeA or AMP, the polymeric nanomaterial subject to the invention can be used as a selective material in commercial diagnostic and biosensor systems.

[0023] The subject matter of the invention is a molecularly imprinted polymeric nanomaterial, said methamphetamine or amphetamine imprinted nanomaterial contains 25-100 pL L-tryptophan functional monomer, 5-5pL ACN (Acetonitrile), 0.25-1 mL HEM(2-Hydroxy ethyl methacrylate), 0.10-0.50 mL ETGMA, 0.02-0.04 mg APS (ammonium persulfate). MeA or AMP in the range of 0.1-1 mg / mL is also used. In an embodiment of the invention, said material comprises 80 pL L-tryptophan functional monomer, 10 pL ACN (Acetonnitrile), 0.5 mL HEM(2-Hydroxy ethyl methacrylate), 0.25 mL ETGMA, 0.03 mg APS (ammonium persulfate), 0.5 mg / mL MeA or AMP.

[0024] The synthesis method of a polymeric nanomaterial subject to the invention that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale comprises the process steps of:

[0025] i. Preparing the pre-complex (with L-tryptophan functional and MeA or AMP) (incubated at 25°C for durations in the range of 30-90 min),

[0026] ii. Microemersion polymerization (under nitrogen gas) of HEM, ETGMA, APS, and pre-complex,

[0027] iii. Washing with a water-ethanol mixture to remove impurities and remove unreacted monomers,

[0028] iv. Desorption of mold molecule from AMP-imp-p(HEM-NMT) and MeA-imp- p(HEM-NMT) nanoparticles by methanokacetonitrile solution,

[0029] v. Separating nanoparticles from the washing medium by centrifugation at 10000- 15000 rpm for 15-30 minutes

[0030] vi. Redispersing desorbed nanoparticles in deionized water and storing at 4°C.In another embodiment of the invention, the synthesis method of a polymeric nanomaterial subject to the invention that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale comprises the process steps of:

[0031] i. Preparing the pre-complex (with L-tryptophan functional and MeA or AMP) (at 25°C temperature for 1 hour),

[0032] ii. Microemersion polymerization (under nitrogen gas) of HEM, ETGMA, APS, and pre-complex,

[0033] iii. Washing with a water-ethanol mixture to remove impurities and remove unreacted monomers,

[0034] iv. Desorption of mold molecule from AMP-imp-p(HEM-NMT) and MeA-imp- p(HEM-NMT) nanoparticles by methanokacetonitrile solution,

[0035] v. Separating nanoparticles from the washing medium by centrifugation at 14100 rpm for 20 minutes

[0036] vi. Redispersing desorbed nanoparticles in deionized water and storing at 4°C.

[0037] In the invention, L-tryptophan was used as the functional group with affinity for MeA and AMP. The reason for using L-tryptophan in the functional monomer structure is that the template molecules MeA and AMP have a hydrophobic structure due to their benzene ring structure and methyl groups. This allows the mold molecules to form secondary interactions; these interactions can be hydrogen bonds, hydrophobic interactions and Van Der Waals bonds. Therefore, when selecting a functional monomer, the hydrophobic properties of NMT and its potential for hydrogen bond formation were considered. In order for L-tryptophan to be included as a monomer in the synthesized p(HEM) structure, the functional monomer NMT (N-methacryloyl-(L)-tryptophan methyl ester) was synthesized. NMT functional monomer was added to the template molecule in Acetonitrile (ACN) to form a pre-complex. The pre-complex was polymerized in the presence of HEM(2-Hydroxy ethyl methacrylate) and ETGMA(Ethylene glycol di methacrylate) and with APS (ammonium persulfate). MeA-imp-p(HEM-NMT) (Methamphetamine imprinted poly(2-hydroxyethyl methacrylate - N-methacryloyl-(L)-tryptophan methyl ester) and AMP-imp-p(HEM-NMT) (amphetamine imprinted poly(2-hydroxyethyl methacrylate - N-methacryloyl-(L)-tryptophan methyl ester) polymers were obtained. The polymers were washed with ethanol-water mixture to remove impurities. Methanol :acetic acid:water was used as desorption agent toremove the mold molecule. The resulting polymers contain cavities suitable for MeA and AMP.

[0038] In the preparation of the pre-complexes, NMT functional monomer, whose optimum values were determined, was added into MeA and AMP in acetonitrile (ACN). HEM (2-Hydroxy ethyl methacrylate) monomer, whose optium value was determined, and ETGMA (Ethylene glycol dimethacrylate) used as a crosslinker were added to the solution obtained after pre-complexation and mixed. APS (ammonium persulfate) was added as an initiator. The monomer mixtures were kept under polymerization conditions for the complete formation of MeA-imp-p(HEM-NMT), AMP-imp-p(HEM-NMT) polymers. As a control group, MeA and AMP-non-imprinted p(HEM-NMT) nanoparticles (NIP-NP) were synthesized using the same recipe. The bulk structure of MeA-imp-p(HEM-NMT) and AMP-imp-p(HEM-NMT) nanoparticles and the presence of functional groups were investigated by Fourier transform infrared spectroscopy (FTIR) (Fig. 1). In FTIR spectra (MeA-imp-p(HEM-NMT) corresponds to the H-bond between methamphetamine (NH) and NMT (OH) extending to the broad peak at 3150-3400 cm-1. The symmetric and asymmetric CH3, CH2and CH bands of NMT at 2850, 2950 and 2991 cm-1 reveal methyl C-H extending to a low peak. Additionally, the weak peaks in the symmetric and asymmetric CH3and CH2bands of methamphetamine between 2900 - 3100 cm-1indicate that it binds with amphetamine-NMT, and that the amount of NMT is higher than that of methamphetamine. The C=O stretching vibrations at 1740 cm-1 and the band at 1386 and 1340 cm1belonging to NMT are attributed to the stretching vibration of the hydroxyl group, aliphatic C=C skeletal vibrations at 1630 cm1and aromatic C=C skeletal vibrations at 1660 and 1452 cm1. The stretching vibration at 1144 cm’1reveals the C-O-C in the ester group. The bands at 650-1000 cm1belong to C-C, C-N and C-0 groups. These peaks indicate that methamphetamine successfully binds with NMT. It cannot be said that there is a significant difference between the M1 , M2, and M3 spectra. However, in M3, amphetamine C-H bands and the band belonging to the H-bond are more prominent, indicating that methamphetamine binds more successfully with NMT.

[0039] In FTIR spectra (AMP-imp-p(HEM-NMT), the broad peak extending from 3150 to 3400 cm’1corresponds to the H-bond between amphetamine (NH2) and NMT (OH). The symmetric and asymmetric CH3, CH2and CH bands of NMT at 2850, 2950 and 2991 cm’1reveal methyl C-H extending to a low peak. Additionally, the weak peaks in thesymmetric and asymmetric CH3and CH2bands of amphetamine between 2900 - 3100 cm-1 indicate that it binds with amphetamine-NMT, and that the amount of NMT is higher than that of amphetamine. The C=O stretching vibrations at 1740 cm-1 and the band at 1386 and 1340 cm-1 belonging to NMT are attributed to the stretching vibration of the hydroxyl group, aliphatic C=C skeletal vibrations at 1630 cm-1 and aromatic C=C skeletal vibrations at 1660 and 1452 cm-1. The stretching vibration at 1144 cm-1 reveals the C-O-C in the ester group. The bands at 650-1000 cm1belong to C-C, C-N and C-0 groups. These peaks indicate that amphetamine successfully binds with NMT. It cannot be said that there is a significant difference between the A1, A2, and A3 spectra. However, in A3, amphetamine C-H bands and the band belonging to the H-bond are more prominent, indicating that amphetamine binds more successfully with NMT.

[0040] Zeta potential and size analysis conducted revealed that MeA-imp-p(HEM-NMT) has a size of 671.2 nm and a potential of -3.1 mV, while AMP-imp-p(HEM-NMT) has a size of 964.9 nm and a potential of -2.3 mV (Fig. 2 and 3). The zeta potentials of AMP-imp-p(HEM-NMT) and MeA-imp-p(HEM-NMT) polymers were -2.3 and -3.1 mV, respectively, providing a suitable surface area for more carboxyl and hydroxyl groups on HEM. Scanning electron microscopy (SEM) was used to determine the surface morphology and bulk properties of MeA-imp-p(HEM-NMT) and AMP-imp p(HEM-NMT) nanoparticles. When SEM results from different distances are examined, it is understood that the particles are spherical and their sizes are compatible (Fig. 4). The surface morphology of the polymers was determined by SEM (Fig. 4). The NP shape has been confirmed as almost spherical. The surface topography is shown in Fig. 5 with AFM imaging. MeA-imp-p(HEM-NMT) and AMP-imp-p(HEM-NMT) generally have a rough surface topography and their Rq (root mean square roughness) values were estimated to be 9.7 and 5.5 nm, respectively.

[0041] Adsorption studies and optimizations were carried out to determine the unit analyte capacity per unit polymer of the polymeric nanomaterials subject to the invention. To obtain calibration solutions, 5 different values in the concentration range 0.25-1.0 mg / mL were used. Absorption readings were recorded at 217 nm for MeA and AMP and calibration curves were generated (Fig. 6). After optimization of adsorption studies in aqueous solutions, the selectivity performances of MeA-imp-p(HEM-NMT), AMP-imp_p(HEM-NMT) nanoparticles were investigated. For this, AMP (amphetamine) forMeA-imp-p(HEM-NMT) and MA (methamphetamine) for AMP-impjo(HEM-NMT) were chosen as competitive analytes. As a result, the selectivity performances of MeA-imp-p(HEM-NMT), AMP-imp-p(HEM-NMT) nanoparticles are given in Fig. 6. Analysis of the graphs revealed that in the presence of AMP, the MeA-imp-p(HEM-NMT) nanoparticle exhibited a relatively low interference signal (relative % Q: 1.27). Likewise, in the presence of MeA, the AMP-imp-p(HEM-NMT) polymer exhibited a low interference rate (relative Q %: 2.46) (Fig. 7). According to the results obtained, the two polymeric nanomaterials synthesized have high affinity and selectivity to distinguish between MeA and AMP.

[0042] The material subject to the invention enables the separation of the analyte with high affinity by forming a single chromatographic column. Thus, it can be used both to obtain high-purity analysis as a result of licensed production and as a selective element in "ready-to-use kit" systems, and it can be actively used in kit systems that can be easily applied at the scene and to biological samples.

[0043] Industrial Applicability of the Invention

[0044] The invention relates to a polymeric nanomaterial that has specific affinity for MeA (methamphetamine) or AMP (amphetamine-derived synthetic drugs) and can exist at the nanoscale, as well as to the synthesis method thereof, and is industrially applicable.

[0045] The invention is not limited to the above descriptions and the person skilled in the art can readily present other different embodiments of the invention. These should be considered within the protection scope of the invention claimed by the claims.REFERENCES

[0046] [1] Dunya Saglik Orgutu, (1994), Lexicon of Alcohol and Drug Terms, Geneva: World Health Organization.

[0047] [2] Blanckaert, Peter; van Amsterdam J.; Brunt T.; van den Berg J.; Van Durme F.; Maudens K. and van Bussel J., (2013), “4-Methyl-amphetamine: A Health Threat For Recreational Amphetamine Users”, Journal of Psychopharmacology, C. 27, S. 9, s.817-822.

[0048] [3] United Nations Office on Drugs and Crime, (2016), World Drug Report, Vienna: United Nation

Claims

CLAIMS1. A molecularly imprinted polymeric nanomaterial, characterized in that it comprises L-tryptophan functional monomer, ACN (Acetonitrile), HEM (2- Hydroxy ethyl methacrylate), 0 ETGMA (Ethylene glycol dimethacrylate), APS (ammonium persulfate), MeA or AMP.

2. The molecularly imprinted polymeric nanomaterial according to claim 1, characterized in that it comprises 25-100 pL L-tryptophan functional monomer, 5-5pL ACN (Acetonitrile), 0.25-1 mL HEM(2-Hydroxy ethyl methacrylate), 0.10- 0.50 mL ETGMA, 0.02-0.04 mg APS (ammonium persulfate), MeA or AMP in the range of 0.1-1 mg / mL.

3. The molecularly imprinted polymeric nanomaterial according to claim 2, characterized in that it comprises 80 pL L-tryptophan functional monomer, 10 pL ACN (Acetonnitrile), 0.5 mL HEM (2-Hydroxy ethyl methacrylate), 0.25 mL ETGMA, 0.03 mg APS (ammonium persulfate), 0.5 mg / mL MeA or AMP.

4. The molecularly imprinted polymeric nanomaterial according to any one of the preceding claims for use as a selective material in commercial diagnostic and biosensor systems.

5. A synthesis method of a molecularly imprinted polymeric nanomaterial, characterized in that it comprises the process steps of:i. Preparing the pre-complex (with L-tryptophan functional and MeA or AMP) (incubated at 25°C for durations in the range of 30-90 min), ii. Microemersion polymerization (under nitrogen gas) of HEM, ETGMA, APS, and pre-complex,iii. Washing with a water-ethanol mixture to remove impurities and remove unreacted monomers,iv. Desorption of mold molecule from AMP-imp-p(HEM-NMT) and MeA- imp-p(HEM-NMT) nanoparticles by methanol :acetonitrile solution, v. Separating nanoparticles from the washing medium by centrifugation at 10000-15000 rpm for 15-30 minutesvi. Redispersing desorbed nanoparticles in deionized water and storing at4°C.

6. The method according to claim 5, characterized in that it comprises the process steps of:i. Preparing the pre-complex (with L-tryptophan functional and MeA or AMP) (at 25°C temperature for 1 hour),ii. Microemersion polymerization (under nitrogen gas) of HEM, ETGMA, APS, and pre-complex,iii. Washing with a water-ethanol mixture to remove impurities and remove unreacted monomers,iv. Desorption of mold molecule from AMP-imp-p(HEM-NMT) and MeA- imp-p(HEM-NMT) nanoparticles by methanol :acetonitrile solution, v. Separating nanoparticles from the washing medium by centrifugation at 14100 rpm for 20 minutesvi. Redispersing desorbed nanoparticles in deionized water and storing at4°C.

7. The molecularly imprinted polymeric nanomaterial synthesized by a method according to any one of claims 5 or 6.

8. The use of a molecularly imprinted polymeric nanomaterial according to claim 7 as a selective material in commercial diagnostic and biosensor systems.

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