Vitamins Quantitative Analysis Service
Submit Your InquiryVitamins are a class of trace organic substances that humans and animals must obtain from food to maintain normal physiological functions and play an important role in human growth, metabolism and development. Depending on their chemical structure and water solubility, vitamins are divided into two categories: water-soluble vitamins and fat-soluble vitamins.
Accurate determination of vitamin content plays an important role in controlling food nutrition and safety. Determination of vitamins in the livestock industry also helps to develop better nutritional feeds and to explore disease, metabolism and other related mechanisms. In addition, vitamin analysis is also needed to study the mechanisms of various human diseases and to enable drug development.
Some vitamin samples may have complex matrices, including fats, emulsifiers, proteins, sterols and other fat-soluble substances, and some vitamins are unstable and require special handling. Creative Proteomics has optimized pre-treatments and assays for different types of vitamins. We have developed highly sensitive, selective and accurate LC/MS/MS analytical methods for the identification and accurate quantification of vitamins in complex matrices.
Service Workflow
Instrument platform: AB Sciex QTRAP® 6500 LC-MS/MS
List of Partial Detectable Vitamins at Creative Proteomics
| Water-soluble vitamins | Vitamin B family: vitamin B1, vitamin B2, vitamin B5, vitamin B6, vitamin B12, folic acid, niacin, pantothenic acid, biotin |
| Vitamin C | |
| Fat-soluble vitamins | Vitamin A, Vitamin D, Vitamin E, Vitamin K |
Application of Vitamins Quantitative Analysis
- Applications in multiple fields such as agronomy, medical physiology, and nutrition
- Research on disease control mechanism
- Analysis of plant vitamin metabolism and regulation
- Research on plant vitamin biosynthesis pathways
- Analysis of bioactive functions (antioxidant, antitumor)
Sample Requirements
Acceptable sample types: animal and plant tissues, cells, blood, etc.
- Blood/plasma: ≥ 500ul/sample
- Urine: ≥ 1ml/sample
- Tissue: ≥ 200mg/sample
- Cells: ≥ 1x107/sample
- Feces: ≥ 500mg/sample
It is recommended to repeat more than six. Large pieces need to be cut into 1cm3 pieces.
Shipment condition: dry ice.
Report Delivery
- Experimental procedure
- Parameters of liquid chromatography and mass spectrometry
- Raw data and quality control of liquid chromatography and mass spectrometry
- Metabolite quantification data
- Custom analysis report
Creative Proteomics has leading-edge technology and best-in-class R&D and manufacturing facilities to help you plan, conduct and report metabolomics studies, making your upstream and downstream experiments seamless. If you have any questions or specific requests, please feel free to contact us.
Case 1. Exploring Vitamin D3 Biosynthesis and Response to UV Treatment in Various Plant Species Using LC-APCI-MS/MS
Background:
The study aimed to investigate the presence of vitamin D3 and its precursors in various plant species and assess the impact of UV treatment on their levels. The biosynthesis of vitamin D3 in plants, its potential as a dietary source, and the need for accurate analytical methods were the key motivations for this research
Samples:
A diverse set of plant materials, including Spinacia oleracea L., Solanum glaucophyllum Desf., Solanum lycopersicum L., Sorghum bicolor (L.) Moench, Capsicum annuum L., and Pisum sativum L., were selected for analysis. The study also utilized a reference material, Spinacia oleracea L., as a benchmark.
Methods:
The research employed a comprehensive methodology that involved multiple steps. The following details outline the technical process:
Sample Preparation: The freeze-dried plant material underwent saponification using potassium hydroxide, ethanol, and ascorbic acid to liberate free forms of vitamin D and sterols. An antioxidant (ascorbic acid) and nitrogen flushing were used to prevent oxidation. Silica solid phase extraction was employed to purify the extracts and eliminate interfering substances.
Chromatographic Separation: High-performance liquid chromatography (HPLC) was performed using a Phenomenex Kinetex PFP column with a gradient elution method. The column's selectivity, coupled with selected reaction monitoring (SRM) on a triple quadrupole mass spectrometer, enabled the separation and quantification of analytes.
Mass Spectrometry Analysis: Atmospheric pressure chemical ionization (APCI) was used for mass spectrometry in positive mode. Deuterium-labeled vitamin D3 served as an internal standard for both vitamin D3 and sterols. SRM was employed for quantification, offering improved selectivity and sensitivity. APCI was chosen over ESI due to higher sensitivity for vitamin D and sterols.
Method Validation: The developed method was thoroughly validated for accuracy, linearity, and reproducibility. Blank samples and internal standards were used to assess selectivity and LOD. Standard addition was used to evaluate accuracy, and inter-day reproducibility was determined across different spiking levels.
Results
The study's results indicated varying levels of cholesterol across different plant species, unaffected by UV treatment. Vitamin D3 and its precursor, 7-dehydrocholesterol, were detected in specific plants following UV exposure, suggesting a photolytic reaction akin to animals. However, vitamin D3 synthesis was not uniform among all plant species studied.
In conclusion, the study successfully developed and validated a rapid LC–APCI-MS/MS method for analyzing vitamin D3 and sterols in plant materials. It shed light on the potential biosynthesis of vitamin D3 in plants and its responsiveness to UV treatment, providing a foundation for further research in this area.
Examples of APCI MS2 scans and product ions scans
Reference
- Jäpelt, Rie Bak, et al. "LC–MS/MS with atmospheric pressure chemical ionisation to study the effect of UV treatment on the formation of vitamin D3 and sterols in plants." Food chemistry 129.1 (2011): 217-225.
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