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Hypercut

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Hypercut

Conclusion on the results of the study sample drug Hypercut

Director of the Federal State Unitary Enterprise "Research Institute of Hygiene, Occupational Pathology and Human Ecology" of the Federal Medical-Biological Agency, doctor of medical sciences, professor.

V.R. Rembovsky

2017

  The purpose of the research: the identification of the pharmaceutical Hypercut.

  Samples provided by: Envenom Pharm.  

  Research Object: Dosage Form - Capsules  

  Components to be determined: L-Carnitine ((3K) -3-hydroxy-4-trimethylammonium butanoate), piperine (bioperin, 1 - piperylpiperidine), yohimbine (methyl ester 17ɑ - hydroxy-yohimban-16ɑ - carboxylic acid), synephrine (4- [1-hydroxy-2- (methylamino) ethyl] phenol), DMAA (methylhexanamine).  

  Research Method: High performance liquid chromatography with mass spectrometric detection in high resolution mode.  

  Measurement tools: High Resolution HPLC-MS/MS by Thermo Scienti fi c (USA), Dionex Ultratimate 3000 chromatograph and Q Exactive mass spectrometer (HESI electrospray - II B positive ionization mode).  

  Brief description of the sample submitted for analysis
  Trade name of the drug: Hypercut. Dosage form - capsules. &Nbsp;

  Appearance of the drug: powder in a green capsule.  

  Active Ingredients:
1) L-carnitine ((3K) -3-hydroxy-4-trimethylammonio-butanoate), gross formula: C 7 H 15 NO 3
2) Piperine (biopsyamine, 1 - piperylpiperidine), gross formula: C 17 H 19 NO 3

3) Yohimbine (17a-hydroxy-yohimban-1ba-carboxylic acid methyl ester), gross formula: C 21 H 26 N 2 O 3

4) Synephrine (4- [1-hydroxy-2- (methylamino) ethyl] phenol), gross formula: C 9 H 13 NO 2
 
5) DMAA (methylhexanamine). gross formula: C 7 H 17 N.  

  Sample Preparation for HPLC Analysis
The capsule was dissolved in 5 ml of methanol (pure for HPLC). The prepared solution was diluted with methanol 100 times and analyzed with 5 µl by HPLC-MS high resolution. &Nbsp;

  Equipment and conditions for carrying out HPLC-MS analysis
Liquid chromatograph UltiMate 3000, with a mass selective detector Q Exactive. Ionization mode: electrospray ionization. The chromatograph is equipped with a Zorbax SB-C8 column with a length of 15 cm, an inner diameter of 4.6 mm, and a particle size of 1.8 μm. The gradient elution mode: eluent A - 0.1% solution of formic acid in water, eluent B - acetonitrile. The ratios of the components of the mobile phase are presented in Table 1. The volumetric rate of the mobile phase through the column is 0.4 ml/min. The temperature of the column thermostat is 35 ° C. The volume of sample injected for analysis 5mkl.

  Table 1. The ratio of the components of the mobile phase

  Time   % B
0 ten
five ten
20 90
40 90
40.1 ten
45 ten
 
 The operating conditions of the mass-selective detector: the flow of the gas-desiccant 45 cu The auxiliary gas flow is $ 12. 35 psi spray pressure. The temperature of the gas desiccant 350 ° C. The temperature of the auxiliary flow 400 ° C. The voltage on the capillary is 3500B. Detection in full ion current scan mode (SCAN): registration of ions in the m/z range from 100 to 1500 in positive ionization. &Nbsp;

  Identification of the components of a sample of the drug by HPLC-MS high resolution
The components were identified by the exact mass of the corresponding protonated molecular ion [M + H] +.

  L-Carnitine Identification
Figure 1 shows the mass chromatogram and the mass spectrum obtained by analyzing a sample solution of a Hypercut sample using an HPLC-MS method reconstructed from the exact mass of L-carnitine [M + H] + - 162.11247.

  Figure 1. Mass spectrum and mass chromatogram of sample solution of Hypercut sample reconstructed by exact mass of L-carntin [M + H] + - 162.11247

  hypercut_1.svg
 

A chromatographic peak with a retention time of 4.34 min corresponds to the compound to be detected. The experimentally recorded mass of 162.1125 deviation from the theoretical is less than 0.1 ppm. Measurement error of the exact mass and elemental composition of the ion (the difference between the measured mass and the calculated one) < 5 ppm, which indicates a reliable identification.

  Piperine Identification
Figure 2 shows the mass chromatogram and the mass spectrum obtained by analyzing the sample solution of the Hypercut sample by HPLC-MS, reconstructed according to the exact mass of pi