Stachyose Analysis Service

Stachyose, a tetrasaccharide consisting of two galactose units, one glucose unit, and one fructose unit, is commonly found in various plant sources such as legumes, seeds, and tubers. Despite being indigestible by humans due to the lack of specific enzymes, stachyose serves as a crucial component in the diet of ruminant animals and plays a significant role in their digestive processes. Additionally, stachyose exhibits prebiotic properties, promoting the growth of beneficial gut bacteria and contributing to digestive health.

Analyzing stachyose content in food and feed products is essential for nutritional labeling, quality control, and regulatory compliance. Moreover, understanding the stachyose content in agricultural crops can aid in crop improvement programs aimed at enhancing nutritional value and overall yield. Additionally, research into stachyose metabolism and its impact on human health requires precise analytical methods for accurate quantification and characterization.

Our comprehensive stachyose analysis services at Creative Proteomics are tailored to accommodate the diverse needs of our esteemed customers, ranging from food industries to pharmaceutical companies and academic research institutions.

Stachyose Analysis Services in Creative Proteomics

Stachyose Quantification: Accurate measurement of stachyose levels in various sample types using high-performance liquid chromatography (HPLC) or mass spectrometry (MS) methods.

Stachyose Structure Elucidation: Characterization of stachyose and its related compounds through advanced analytical techniques such as nuclear magnetic resonance (NMR) spectroscopy and tandem mass spectrometry (MS/MS).

Stachyose Content Determination: Determination of the stachyose content in complex matrices, including food products, plant extracts, and biological samples.

Stachyose Purity Analysis: Assessment of the purity of stachyose preparations using chromatographic methods to ensure product quality and consistency.

Stachyose Degradation Studies: Investigation of the stability and degradation pathways of stachyose under various storage and processing conditions.

Customized Stachyose Analysis: Tailored analytical solutions to address specific research questions or project requirements related to stachyose analysis.

Technical Platforms for Stachyose Analysis

High-Performance Liquid Chromatography (HPLC): Our analytical approach capitalizes on HPLC systems featuring precision pumps, autosamplers, and state-of-the-art detectors (UV, RI, or ELSD). These systems afford exceptional separation efficiency and quantification accuracy, facilitating the meticulous characterization of Stachyose within complex matrices.

Mass Spectrometry (MS): Leveraging both liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS), we deploy high-resolution analyzers and tandem mass spectrometry configurations. These platforms empower sensitive detection and structural elucidation of Stachyose and its congener compounds, ensuring comprehensive characterization across diverse sample types.

Nuclear Magnetic Resonance (NMR) Spectroscopy: Our investigative arsenal encompasses cutting-edge NMR spectrometers, augmented by formidable magnetic fields and advanced pulse sequences. This suite of instrumentation facilitates the elucidation of Stachyose's molecular architecture and the authentication of its compositional integrity.

Customized Analytical Solutions: Through an iterative process of methodological refinement, our adept team tailors analytical protocols to the nuanced demands of specific research inquiries and sample matrices. This bespoke approach ensures optimal performance characteristics and data fidelity, thereby empowering nuanced insights into Stachyose's compositional dynamics.

Sample Requirements for Stachyose Analysis

Applications of Stachyose Analysis

Food science: Assessing stachyose content in food products for nutritional labeling and quality control.

Pharmaceuticals: Investigating stachyose in herbal extracts for pharmacological research and formulation development.

Agriculture: Studying stachyose in plant physiology and its role in symbiotic relationships and stress responses.

Gut microbiota research: Analyzing stachyose fermentation by gut microbiota to understand its impact on human health.

Functional ingredient development: Identifying stachyose as a potential functional ingredient for formulating health-promoting products.

Biomedical studies: Examining stachyose's physiological effects and therapeutic potential in metabolic and immune disorders.

Animal Nutrition: Assessing stachyose content in animal feed for optimizing nutritional formulations and promoting livestock health.

Environmental Science: Studying stachyose as a marker for environmental contamination and its degradation pathways in soil and water systems.

Functional Genomics: Analyzing stachyose metabolism-related genes and pathways to elucidate its biosynthesis and regulatory mechanisms in plants and microorganisms.

Title: LC-MS/MS Method Development for Stachyose Quantification in Rat Plasma and Pharmacokinetic Study of Micronized RGOS

Background

Stachyose, a functional oligosaccharide found in legumes and traditional Chinese medicine, holds promise for various therapeutic applications due to its prebiotic properties and potential health benefits. However, its pharmacokinetic profile following pulmonary administration of micronized RGOS remains unclear, necessitating the development of a sensitive analytical method for quantification.

Sample

Rat plasma samples were utilized for pharmacokinetic analysis after pulmonary administration of micronized RGOS containing stachyose. The samples underwent protein precipitation for sample preparation to extract stachyose for analysis.

Technical Methods

LC-MS/MS Condition Optimization: The electrospray ionization (ESI) conditions for stachyose and the internal standard (IS) were optimized using quadrupole full scan mass spectra in both positive and negative detection modes. Negative ion mode showed higher sensitivity, and precursor/product transition pairs (m/z [M-H]- 665.4→383.1 for stachyose and 665.5→485.0 for IS) were selected for quantification.

Chromatographic Separation: Various mobile phase compositions including methanol-water, acetonitrile-water, and those containing 0.05% formic acid were tested. The best separation and peak shape were achieved with acetonitrile-water (55:45, v/v). An NH2 chromatographic column (Asahipak NH2P-50 4E) coupled with a guard column (Asahipak NH2P-50G 4A) was used to ensure good peak shape and short retention time.

Sample Preparation: Protein precipitation was employed for sample preparation due to stachyose's high hydrophilicity. Acetonitrile was used as the precipitation agent, resulting in low matrix effect and satisfactory extraction recovery. Rat plasma samples were thawed, and 100 µl was mixed with 100 µl of acetonitrile containing IS (2 µg/ml). After vortexing and centrifugation, the supernatant was injected into the LC-MS/MS system.

Method Validation: The method was validated according to US FDA guidelines. Specificity was confirmed by analyzing blank plasma samples, spiked plasma samples, and plasma samples after administration. The calibration curve showed linearity over the concentration range of 100-30000 ng/ml, with an LLOQ of 100 ng/ml. Precision and accuracy were within acceptable limits, and extraction recovery and matrix effect were stable. Stachyose stability was assessed under various conditions, demonstrating stability in rat plasma and solution storage.

Pharmacokinetic Study: Male Sprague-Dawley rats underwent pulmonary administration of micronized RGOS containing stachyose. Blood samples were collected at various time points, and plasma concentrations were determined using the validated LC-MS/MS method. Pharmacokinetic parameters were calculated using noncompartmental analysis software, revealing rapid absorption and elimination of stachyose following pulmonary administration of micronized RGOS.

Results

The developed LC-MS/MS method exhibited satisfactory specificity, linearity, accuracy, precision, extraction recovery, matrix effect, and stability. Specificity was confirmed by representative chromatograms, and the calibration curve showed linearity over the concentration range of 100-30000 ng/ml with an LLOQ of 100 ng/ml. Pharmacokinetic analysis revealed rapid absorption of stachyose with a Tmax of 0.2 h and fast elimination with a T1/2 of 3.5 h following pulmonary administration of micronized RGOS.

Full–san product ion spectra of [M-H]- for (A) stachyose and (B) nystose (IS).

Representative LC-MS/MS chromatograms for stachyose and IS in rat plasma

Reference

1. Zhou, Yang, et al. "A LC–MS/MS method for the determination of stachyose in rat plasma and its application to a pharmacokinetic study." Journal of Pharmaceutical and Biomedical Analysis 123 (2016): 24-30.