A novel approach to the analysis of multivitamin by online supercritical fluid extraction/supercritical fluid chromatography

Importance of the Topic

Fat soluble vitamins are essential micronutrients for animal and human health. Traditional analytical workflows rely on multiple methods for different vitamins, increasing time and resource costs. An integrated online supercritical fluid extraction and chromatography approach offers rapid, reliable and automated quantification of vitamins A, D and E in pharmaceuticals and health foods.

Objectives and Study Overview

This study aimed to develop a single analytical protocol for five fat soluble vitamins (vitamin A acetate, vitamin A palmitate, vitamin E acetate, vitamin D2 and vitamin D3) using an online SFE-SFC system. The primary goals were to unify sample pretreatment, improve throughput and ensure accurate quantification in commercial formulations.

Methodology and Instrumentation

Sample Preparation:

• Standard solutions of five vitamins prepared in n-hexane and ethanol to generate calibration curves
• Commercial tablets, capsules and pearls loaded directly into extraction vessel

Instrumentation:

• Nexera UC Online SFE-SFC system equipped with SFE-30A module, LC-30ADSF CO2 pump, LC-20ADXR modifier pump, degasser DGU-20A5, column oven CTO-20AC, back pressure modules SFC-30A, UV detector SPD-20A and LabSolutions Ver5.8 workstation

SFE Conditions:

• Supercritical CO2 with 5% methanol modifier at 5 mL/min
• Static extraction for 3 min at 50 C and 14.8 MPa, followed by dynamic extraction for 3 min

SFC Conditions:

• ODS column 4.6x250 mm 5 um
• Gradient elution from 0% to 50% methanol modifier over 17 min
• Flow rate 3 mL/min, column oven 40 C, backpressure 10 MPa
• Detection wavelengths 325 nm for vitamins A and D, 284 nm for vitamin E

Main Results and Discussion

Online SFE achieved over 85% extraction efficiency in a single dynamic step, with above 94% recovery in first pass. Chromatographic separation resolved all five analytes within 15 min. Calibration curves were linear over relevant concentration ranges (R2 0.997–0.999). Repeatability (n=6) showed peak area RSD below 13% and retention time RSD below 0.25%. Recoveries in fortified samples ranged from 90% to 101%. Analysis of real commercial samples yielded clear, well separated peaks and accurate concentration values.

Benefits and Practical Applications

Combining SFE and SFC in an online workflow removes offline pretreatment, reduces solvent use and shortens overall analysis time. Automation improves reproducibility and lab throughput. This method is well suited for pharmaceutical quality control, nutraceutical testing and regulatory compliance in multivitamin formulations.

Future Trends and Potential Applications

Coupling online SFE-SFC with mass spectrometry could extend analyte coverage to include additional lipid soluble compounds. Miniaturized extraction vessels and greener modifiers will enhance sustainability. Integration with real-time data analytics may enable continuous manufacturing quality control for complex dosage forms.

Conclusion

A fully automated online SFE-SFC method was established for simultaneous quantification of vitamins A, E and D in drug and health food matrices. The approach offers high efficiency, robust performance and broad applicability.

Reference

• Qiang Li Hongyuan Hao Taohong Huang Shin-ichi Kawano Yuki Hashi A novel approach to the analysis of multivitamin by online supercritical fluid extraction supercritical fluid chromatography Pittcon 2016 PO-CON1618E Shimadzu China Co Ltd 2016