Fudan University Journal of Medical Sciences

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Reverse phase high-performance liquid chromatography (RP-HPLC) system for analysis of vitamin D3 soft capsules with soybean oil matrix

LI Meng-han▲, DING Xue-mei▲, LIANG Jian-ying△

jms, 2019, 46(01): 23-31. DOI: 10.3969/j.issn.1672-8467.2019.01.004

A:Thermal degradation; B:Photo degradation; C:Thermal and photo degradation.Peak 1:Trans-vitamin D3;Peak 2:Pre-vitamin D3;Peak 3:Tachysterol; Peak 4:Vitamin D3. Chromatograms of forced degradation samples

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R:-CHH₃(CH₂)₃CH(CH₃)₂ Photochemical and thermal isomers of vitamin D3
a:Vitamin D3 (λ_max=262 nm); b:Pre-vitamin D3 (λ_max=260 nm); c:Trans-vitamin D3 (λ_max=276 nm); d:Tachysterol (λ_max=280 nm).λ_max:Maximum absorption wavelength. UV absorption spectrums of vitamin D3 and its three isomers
1 Effect of mobile phase composition on various chromatographic parameters
A and B:a-e, V (ACH):V (MeOH)=99:1, 98:2, 97:3, 90:10 and 80:20;C:a-c, V (ACH):V (H₂O)=95:5, 90:10 and 85:15.MeOH:Methanol; ACN:Acetonitrile; H₂O:Water. Chromatograms of sample 4 with various proportions of methanol (A), ethanol (B) and water (C) in acetonitrile as mobile phase
a:100%ACN; b-e:V (ACN):V (MeOH)=99:1, 98:2, 97:3, 90:10, respectively; f-i:V (ACN):V (EtOH)=99:1, 98:2, 97:3, 90:10, respectively. Asymmetry of vitamin D3 peak using different mobile phases
6 Effect of ethanol or methanol addition in acetonitrile as mobile phase on recovery of vitamin D3 soft capsules with oil matrix (n=6)
A:100% acetonitrile; B:V (acetonitrile):V (H₂O)=95:5.a:Degradation sample (sample 3);b:Blank matrix of vitamin D3 soft capsule. Chromatograms of degradation sample and blank matrix with 100% acetonitrile and V (acetonitrile):V (H₂O)=95:5 as mobile phase

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