## 2 Materials and Methods
### Vitamin Analysis  
To determine the concentration of plasma vitamin A (retinol), a plasma sample (1 mL) was prepared. Proteins were denatured with 500 \(\mu\)L pure ethanol followed by brief mixing with a vortex (15 s); then the sample was extracted twice with 500 \(\mu\)L of n-hexane. The organic layers were pooled and dried under nitrogen. The dried sample was reconstituted with 100 \(\mu\)L of methanol, vortexed, and centrifuged briefly at 1380\(\times\)\(g\) for 30 s before 35 \(\mu\)L of subsample was injected into an HPLC system (Agilent Technologies Inc., Santa Clara, CA, USA). Absorbance was monitored at 350 nm to maximize the detection of retinol, and a mobile phase of 98:2 methanol:water was run at a flow rate of 1 mL/min. To extract vitamin E (\(\alpha\)-tocopherol) from the plasma, the procedure of van Kempen was used [12]. In brief, plasma samples (1 mL) were extracted by adding 1 mL ethanol containing 0.15% butylated hydroxytoluene. Subsequently, 3 mL n-hexane containing 0.025% butylated hydroxytoluene was added into the extraction. After vortex and centrifuging (180\(\times\)\(g\) for 5 min at 20 \({}^{\circ}\)C), 2 mL of the n-hexane layer was collected, dried using N\({}_{2}\), and resuspended in 200 \(\mu\)L methanol. This extract was analyzed using an Agilent Plastisil C18 ODS, 250 \(\times\) 4.6 \(\mu\)m column (Agilent Technologies Inc., Santa Clara, CA, USA) on an HPLC (Agilent 1200 Series; Agilent Technologies Inc., Santa Clara, CA, USA). The identification and quantification of \(\alpha\)-tocopherol were based on the retention time and peak area of the authentic standards. Plasma 25-hydroxyvitamin D\({}_{3}\) (25(OH)D\({}_{3}\)) concentrations in the blood were determined following the protocol of commercially available ELISA kits (Immunodiagnostic Systems Holdings PLC, London, UK). Vitamin K\({}_{3}\) (Menadione) in blood was determined by LC-MS (DIONEX Ultimate 3000 and Thermo Q EXACTIVE). Next, 50 \(\mu\)L of plasma was added to 2 volumes of methanol-precipitated protein and centrifuged at 13,200 rpm for 4 min; then, the supernatant was taken into a new tube, 450 uL of n-hexane was added into this tube to vortex for 1 min, and then the tube was centrifuged at 13,200 rpm for 8 min. The upper organic phase was placed in a new PE tube and concentrated with N\({}_{2}\). After concentration and drying, this mixture was reconstituted by adding 100 \(\mu\)L of methanol, and 50 \(\mu\)L of the mixture was taken for detection. Plasma thiamine, riboflavin, vitamin B\({}_{6}\), niacin, folic acid, biotin, and pantotheanate concentrations were determined by LC-MS (Shimadzu LC20 AD-API 3200 MD TRAP). we next took 50 \(\mu\)L of the plasma sample directly, added 150 \(\mu\)L of acetonitrile to precipitate the protein, and shook the mixture for centrifugation. Then, 50 \(\mu\)L of the mixture was taken for detection. Concentrations of plasma vitamin B\({}_{12}\) (VB\({}_{12}\)) were measured by LC/MS/MS (Agilent 1260 Series; Agilent Technologies Inc., Santa Clara, CA, USA). In brief, a 200 \(\mu\)L blood sample was prepared, 100 \(\mu\)L of 5% trichloroacetic acid was added, and the mixture was vortexed for 5 min prior  
\begin{table}
\begin{tabular}{c c} \hline
**Items** & **Percent** \\ \hline Trace mineral premix [1] & 0.50 \\ Vitamin premix [2] & 0.03 \\ Total & 100.00 \\ Calculated nutritional values & \\ Metabolized energy (Kcal/kg) & 3455.16 \\ Crude protein & 18.25 \\ SID Lysine & 1.35 \\ SID Methionine & 0.39 \\ SID Threonine & 0.79 \\ SID Trytopphan & 0.22 \\ SID Valine & 0.86 \\ Total Calcium & 0.79 \\ STTD Phosphorus & 0.41 \\ \hline \end{tabular}
\end{table}
Table 3: _Cont._to centrifugation for 10 min at 13,000 rpm. Next, 25 uL of 1 N sodium hydroxide was added, and the supernatant was transferred to MS vials for analysis.