Samples provided by
: Envenom Pharm.
The purpose of the research: the identification and determination of the active principle in the sample prepara-
Object of study:
dosage form - capsules.
Component to be detected:
4 - ((R) -2 - ((R) -2,2,2-trifluoro-1-hydroxyethyl) porrolidin-1-yl) -2-
Date of receipt of the sample for research:
High performance liquid chromatography with mass selective detection -
in high resolution mode.
Acella Liquid Chromatograph with Mass Spectrometry Detector - LTQ
Orbitrap Velos with electrospray ionization at atmospheric pressure and software
Xcalibur data management and processing software.
Identification of 4 - ((R) -2 - ((R) -2,2,2-trifluoro-1-hydroxyethyl) porrolidin-1-yl) -2-
trifluoromethyl) benzonitrile in the sample was performed by high performance liquid chromatography method
Fies with mass selective detection in high resolution mode. Component Separation
samples were carried out on a chromatographic column of a liquid chromatograph with direct detection-
by the separation of separated components by a mass spectrometry detector.
Brief description of the sample submitted for analysis.
Trade name of the drug:
Ligandrol. Synonyms: Ligandrol, LGDRx, LGD-4033. Dosage Form -
capsules containing 10 mg of the active ingredient. The active ingredient is a substance with chemical-
By the name of 4 - ((R) -2 - ((R) -2,2,2-trifluoro-1-hydroxyethyl) porrolidin-1-yl) -2-
Gross formula: С14Н12F6N2O
Molecular Weight: 338.0854 g/mol
Appearance of the preparation: powder in a two-color, green-white capsule.
The active ingredient’s structural formula:
Sample preparation for analysis.
A sample of a 10 mg sample was dissolved in 1 ml of water. For a VYGH-MS analysis, the prepared solution
diluted 100 times.
Chromatographic Separation Conditions.
Chromatography column - Zorbax SB-C8, 150 mm long, 4.6 mm in diameter, particle size
1.8 micron sorbent.
- component A - 0.1% solution of formic acid in deionized water;
- component B - acetonitrile of the category “for gradient VYHH”;
Mobile Phase Ratio
The eluent flow rate is 0.4 ml/min;
Column thermostat temperature: 35 ° С;
The volume of sample injected for analysis of 5 µl.
Operating conditions for a mass spectrometric detector.
Mass spectrometry detector - LTQ Orbitrap Velos with electrospray ionization at
atmospheric pressure and Xcalibur data management and processing software.
- Gas flow desiccant 45 cu
- Auxiliary Gas Flow $ 15
–The temperature of the gas-drying 300 ° C;
–Auxiliary flow temperature 380 ° С;
- The voltage on the capillary 3500V;
–Detecting in Full Ion Flow Scan Mode (SCAN): Ion Registration in
m/z range from 50 to 1000 throughout the analysis (with positive ionization).
The main component was detected by m/z 339.0927, which corresponds positively
+ charged protonated molecular ion [M + H] 4 - ((R) -2 - ((R) -2,2,2-trifluoro-1-hydroxy-e-
Tyl) porrolidin-1-yl) -2-trifluoromethyl) benzonitrile.
Figure 1 - HPLC-MS - chromatogram of the sample solution of the drug "Ligandrol"
and the mass spectrum of the main component.
Chromatographic peak with a retention time of 5.59 minutes corresponds to the current + [M + H] ion
substances of the drug "Ligandrol" with a mass number of m/z 339.09266. Deviation from the theoretical
is less than 0.1 ppm. Measurement error of the exact mass and elemental composition of the ion (difference
measured mass from the calculated) & lt; 5ppm, identification is authentic.
A chromatographic peak with a retention time of 4.28 minutes corresponds to an impurity component.
A quantitative assessment of the main substance was performed by the method of internal normalization. Content
active ingredient in the sample drug "Ligandrol" was 98%.
As a result of the study it was found that in the presented sample of the drug "Ligan-
Drol "the main substance is 4 - ((R) -2 - ((R) -2,2,2-trifluoro-1-hydroxyethyl) porrolidin-1-yl) -2-
trifluoromethyl) benzonitrile. Estimated content of the analyte in the preparation