Application of UPLC-MS/MS Method for Analyzing B-vitamins in Human Milk

Area of Science:

• Analytical Chemistry
• Biochemistry
• Nutritional Science

Background:

• Human milk is a crucial source of nutrition for infants.
• B-vitamins are essential micronutrients vital for infant development.
• Accurate quantification of B-vitamins in human milk is critical for nutritional assessment.

Purpose of the Study:

• To develop and validate a simultaneous analytical method for ten B-vitamins in human milk.
• To utilize ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for precise vitamin determination.
• To establish a reliable method for assessing B-vitamin content in human milk.

Main Methods:

• Human milk samples were pretreated and analyzed using UPLC-MS/MS within 11 minutes.
• A gradient elution with ammonium formate and acetonitrile was employed on an Acquity UPLC HSS T3 column.
• Stable isotope internal standards were used to correct for matrix and ionization effects, with detection via ESI+ ionization in MRM mode.

Main Results:

• The method achieved high linearity (R²=0.990-0.999) across specified concentration ranges for the ten B-vitamins.
• Excellent relative recovery (80.1%-120.2%) and accuracy (98.3%-108.0%) were demonstrated.
• Low limits of quantification (0.25-3 µg/L) and acceptable intra-day (3.4%-19.9%) and inter-day (5.9%-18.1%) variations were achieved.

Conclusions:

• The developed UPLC-MS/MS method provides a robust and efficient means for the simultaneous analysis of ten B-vitamins in human milk.
• This validated method is suitable for routine nutritional monitoring and research involving human milk.
• The findings contribute to a better understanding of B-vitamin composition in human milk.