| Biochemical drugs are biochemical substance which are applied for the prevention, treatment and diagnosis of diseases. Biochemical drugs could be obtained through isolation and purification from organisms, or through chemical synthesis or biotechnology. Phospholipid drugs and peptide drugs are both important component of biochemical drugs. The first part of this research was to analyze the single component in phospholipid drugs qualitatively and quantitatively, and the second part of this research was to study the detection method of acetic acid in polypeptide drugs.In the first part of this research, separation effect of phospholipid component from egg yolk lecithin by Silica gel column, glycol column and Hilic column was observed initially. Hydrophilic interaction chromatography was selected after comparison. Detector, which was used to analyze phospholipid drug was mass spectrum. Afterwards, took egg yolk lecithin, phosphatidylcholine and bovine lung extract as the object of study, exact molecular weight of each phospholipid compound was measured by mass spectrometry for the sake of speculating the element composition, and fracture law was obtained with fragment ions from MS/MS. Finally, the results were validated by searching LipidMaps database. Structural analytic methods of phosphatidyl choline, lysophosphatidylcholine, sphingomyelin, phosphatidyl ethanolamine, lysophosphatidyl ethanolamine, phosphatidyl inositol, phosphatidylglycerol and phosphatidal ethanolamine were summarized. By this means, we found 133 kinds of phospholipid compound in 3 kinds of phospholipid drugs. At last, with Tenofovir eighteen alkoxy ethyl mono ester derivatives as interior standard substance, the content of DPPC in bovine pulmonary surfactant was detected and methodology validation took place subsequently. The experiment results show that The linearly dependent coefficient was satisfied when the content of DPPC ranged from 2.152～21.52μg/ml (r2=0.9992). The average recovery was 107.78% with a RSD of 8.18%. The content of DPPC in 4 batches of bovine pulmonary surfactant were 19.26%, 35.66%,35.89% and 35.07% respectively. The method is sensitive, specific, accurate and precise, which can be applied to determine the content of DPPC in bovine pulmonary surfactant.Objective of the second part of this research is to establish an ion-exclusion chromatography(IEC) method to explore the content of acetic acid in Triptorelin Acetate Injection and Octreotide Acetate for Injection. Rezex ROA-Organic Acid H’(7.8 X 300 mm) column was applied, with the mobile phase being 0.0025 mol/L and sulfuric acid at the flow rate of 0.5 ml/min. The column temperature was 45℃ and detection wavelength was 210 nm. The content of acetic acid was calculated according to peak area by external standard method. The experiment results show that the LOD of acetic acid was 78.89 ng/ml, and the LOQ was 197.2 ng/ml. The linearly dependent coefficient was 0.9999 when the content of acetic acid ranged from 0.3944μg/ml to 78.89μg/ml. The average recovery(n=9) were 98.69% with a RSD of 0.51% and 107.09% with a RSD of 1.34% respectively. The content of acetic acid in 3 batches of Triptorelin Acetate Injection were 23.99μg/ml,22.38μg/ml,22.96μg/ml respectively. The content of acetic acid in 3 batches of Octreotide Acetate for Injection were 2.15%,2.10%,2.08% respectively. This method was environmental, concise, accurate and precise. It could be applied to determine the content of acetic acid in polypeptide drugs. |
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