Injection Preparation And Content Of The Freeze-dried Powder Of The Nine Water-soluble Vitamin Determination

In this paper,the lyophillization of nine water-soluble vitamins for injection was investigated,which included vitamin B₁(VB₁),vitamin B₆(VB₆),vitamin B₁₂(VB₁₂), vitamin C(VC),Nicotinamide,folic acid,D-Biotin,riboflavin 5'-phosphate sodium, D-pantothenic acid sodium salt.Content determination of nine water-soluble vitamins for injection was observed.In the first chapter,we had a research for the influencing factors such as the excipient,viscosity of solution and lyophillization manner on the formula and preparation technology of the lyophilized nine water-soluble vitamins.The results indicted that trehalose(300 mg/ampoule)as supporting agent,4 mL of drug solution in an ampoule and pre-freeze in a repeated way were chosen to make the lyophillization product.In the second chapter,nine vitmins were determined by high performance liquid chromatography(HPLC).The work includes three parts.(1)Simultaneous determination of biotin,folic acid and methyl p-hydroxybenzoate in water-soluble vitamin for injection by HPLCThe HPLC system equipped with a 5μm ODS C₁₈250×4.6 mm column using gradient elution program was adopted.The mobile phase was potassium dihydrogen phosphate buffer - acetonitrile,and the column temperature was 30℃.The flow rate was 1.5 mL/min,and the detection wavelength was 200 nm.The linear ranges of biotin,folic acid and methyl p-hydroxybenzoate were 8.05～24.15μg/mL,1.27～3.81μg/mL,9.95～29.85μg/mL,respectively.In the linear range the correlation coefficients were 0.9991～0.9995.The average recoveries ranged from 96.3%to 104.2%with less than 3.60%of intra-day and inter-day RSD.The detection limits were 0.30μg/mL,1.00μg/mL,0.10μg/mL,respectively(2)Determination of nicotinamide,VB₆,VB₁,D-pantothenic acid sodium salt,VC, riboflavin 5'-phosphate sodium in water-soluble vitamin for Injection by HPLC The HPLC system equipped with a 5μm NH₂ 250×4.6 mm column was adopted. The mobile phase was 0.02 mol/L potassium dihydrogen phosphate buffer-acetonitrile (27:73,V/V),and the column temperature was 30℃.The flow rate was 1.0 mL/min. To simultaneously analyze nicotinamide,VB₆,VB₁,D-pantothenic acid sodium salt, VC,the UV detection wavelength was set at 214 ran.Riboflavin 5'-phosphate sodium was investigated by fluorescence detection.Theλ_exwas 445 nm,andλ_exwas 520 nm. The linear ranges of nicotinamide,VB₆,VB₁,D-pantothenic acid sodium salt,VC, riboflavin 5'-phosphate sodium were 30.22～90.66μg/mL,3.76～11.28μg/mL, 2.38～7.14μg/mL,12.44～37.32μg/mL,85.06～255.18～g/mL,1.94～11.64μg/m,,respectively.In the linear range the correlation coefficients ranged from 0.9992 to 0.9999.The average recoveries were 96.3%-104.2%with less than 3.40%of intra-day and inter-day RSD.The detection limits were 0.15,0.30,0.80,5.00,1.40, 0.15μg/mL,respectively.(3)Determination of VB 12 in water-soluble vitamin for injection by HPLCThe HPLC system was adopted,on a 5μm ODS C₁₈250×4.6 mm column, gradient elution,using the mobile phase of A:phosphate buffer-acetonitrile(90:10, V/V),and B:acetonitrile-water-phosphate acid(499:499:2,V/V/V).The column temperature was 30℃.The detection wavelength was 200 nm.The linear range of VB₁₂was 0.50～1.50μg/mL.In the linear range the correlation coefficients was 0.9996.The average recoveries ranged from 101.0%to 102.5%.The intra-day and inter-day RSD was less than 1.80%.The detection limit was 0.40μg/mL.In the third chapter,nine water-soluble vitamin for injection were simultaneously determined by high performance capillary electrophoresis(HPCE).Micellar electrokinetic capillary chromatography(MEKC)modes was operated.The system was equipped with fused-silica capillary column:67 cm(efficiency length 55 cm)×50μm.The running buffer was 50 mmol/L SDS-50 mmol/L Na₂B₄O₇·10H₂O solution (pH = 8.33).The eletroinjection was 6.89 KPa×l0 sec with 15 kv running voltage. The detection wavelength was 214 nm.The column temperature was 25℃.The linear ranges of VB₁,VB₆,VB₁₂,VC,nicotinamide,folic acid,D-biotin,riboflavin 5'-phosphate sodium,D-pantothenic acid sodium salt were 2.405～19.24μg/mL, 3.8～30.4μg/mL,0.39～3.12μg/mL,87.5～700.0μg/mL,31～248μg/mL,0.31～2.48μg/mL,0.465～3.72μg/mL,3.8～30.4μg/mL,4.125～33.0μg/mL,respectively. In the linear range the correlation coefficients were 0.9982 - 0.9998.The average recoveries ranged from 95.8%to 100.8%.The mean peak area and migration time of intra-day and inter-day RSD was less than 3.30%.The detection limits were 0.18μg/mL,0.16μg/mL,0.10μg/mL,0.45μg/mL,0.60μg/mL,0.10μg/mL,0.12μg/mL, 0.23μg/mL;0.41μg/mL,respectively.Compared with the HPLC method,HPCE was more convenient.In general,the nine water-soluble vitamins for injection were prepared by lyophillization and determined by HPLC and HPCE.This work made a good foundation of the development of this preparation.