Vitamin-like Compounds Analysis Service

What are Vitamin-like Compounds?

There are a variety of organic compounds though cannot be defined as true vitamins, which are related to the function of vitamins. Generally speaking, they can be sufficiently synthesized by humans and are not required in the daily diet. Because of similarities in physiological function or distribution in diet, these substances usually are classified as vitamin-like substances. Besides specific amino acids such as methionine and taurine in feathered animals, other examples of these vitamin-like substances include choline, carnitine, inositol and Coenzyme Q.

Choline

As a key component of sphingomyelin and lecithin, choline, a water-soluble amine, plays an important role in carcinogenesis, lipid transport and methyl group metabolism. Normally, choline can be synthesized in sufficient amounts. However, choline is regularly added to diets, commercially available as the bitartrate or trimethyl hydroxyethyl ammonium chloride. This can lessen the need for activated methyl groups supplied from methionine, and thus promotes positive growth response in young growing animals. Choline deficiency can lead to fatty liver.

Inositol

Inositol is synthesized after cyclization glucose-6-phosphate and is after cyclization and is of similar structure to glucose. As a component of phospholipids in membranes, it plays a key role in the cell replication. Inositol also plays an important role in phospholipid assembly, clearance of lipid and cellular signal transduction. Inositol is relatively abundant in cereal grains. Similar to choline, deficiency of inositol can lead to fatty liver.

Taurine

Taurine is an amino acid taking part in in a variety of physiological activities, including neuromodulation, osmotic regulation and the stabilization of cell membranes. It is essential for the metabolism of bile acids salts. From the oxidation of cysteine, most animals can synthesize sufficient amounts of taurine endogenously. However, some animals, especially domesticated and wild felids and human infants fail to synthesize enough amounts of taurine. Currently, taurine is regularly added to all infant formulas to promote infant development.

Carnitine

Carnitine plays an important role in accepting activated fatty acids at the outer mitochondrial membrane and making them ready for β-oxidation. While adults can synthesize sufficient carnitine, it is difficult for infants to synthesize enough amounts of carnitine. Human milk delivers sufficient carnitine, infant formula may not offer enough carnitine. Carnitine is relatively abundant in meats and dairy products, while cereal grains is not only low in carnitine but also low in lysine and methionine- the precursors of carnitine. The primary signs of carnitine deficiency are as hypoglycemia, cardiomyopathy and muscle weakness.

Bioflavonoids

Bioflavonoids are the brightly colored phenolic compounds relatively abundant in tea, beer, wine, cocoa, and especially in citrus fruits. They can affect capillary permeability and fragility.

Lipoic Acid

As a fat-soluble vitamin relating to B vitamins, lipoic acid acts as a coenzyme to transfer acyl groups.

Coenzyme Q (Ubiquinone)

Coenzyme Q is defined as a group of lipid-like compounds with structure similar to vitamin E. Coenzyme Q plays a key role in mitochondrial electron transport. Coenzyme Q10, the member native to human mitochondria, is of greatest interest. Coenzyme Q is relatively abundant in the food supply.

Vitamin-like Compounds Analysis Offered by Creative Proteomics

Quantitative Analysis: Accurate measurement of vitamin-like compounds, including ascorbic acid, biotin, and various forms of Vitamins B, C, D, E, and K.

Method Development: Custom analytical methods for specific compounds, optimizing techniques like HPLC or LC-MS for enhanced detection and quantification.

Stability Testing: Assessment of vitamin stability under varying conditions (temperature, light, humidity) to determine shelf life and efficacy.

Biological Sample Analysis: Evaluation of biological samples (e.g., serum, plasma) to determine levels of vitamins and assess nutritional status in individuals.

Nutritional Content Evaluation: Analysis of food products to quantify vitamin content, ensuring compliance with nutritional labeling and safety regulations.

Metabolite Profiling: Investigation of metabolic pathways related to vitamins, providing insights into their biological functions and impacts on health.

List of Vitamin-like Compounds We Can Detect

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Technology Platforms for Vitamin-like Compounds Analysis

High-Performance Liquid Chromatography (HPLC): This technique is commonly used for separating and quantifying vitamins such as ascorbic acid, biotin, and riboflavin. HPLC systems like the Agilent 1260 or Shimadzu Prominence are often employed, featuring UV-Vis detectors for sensitive detection.

Liquid Chromatography-Mass Spectrometry (LC-MS): LC-MS is utilized for the comprehensive analysis of complex vitamin profiles, allowing for high specificity and sensitivity. Instruments like the Thermo Scientific Orbitrap or Waters Xevo TQ-S are ideal for detecting low concentrations of vitamins and their metabolites.

Gas Chromatography-Mass Spectrometry (GC-MS): This technique is used for analyzing volatile vitamins and related compounds. Instruments such as the Agilent 7890B GC coupled with the Agilent 5977A MS provide robust analysis capabilities.

Nuclear Magnetic Resonance (NMR) Spectroscopy: NMR is employed for structural elucidation and quantification of vitamin-like compounds. Systems like the Bruker Avance III are used to gain insights into the molecular structure and dynamics of these compounds.

Enzyme-Linked Immunosorbent Assay (ELISA): For specific vitamins, such as Vitamin D, ELISA kits can be used for quantification. Automated systems like the BioTek ELx808 can streamline this process.

Sample Requirements for Vitamin-like Compounds Analysis

Sex Modifies the Impact of Type 2 Diabetes Mellitus on the Murine Whole Brain Metabolome

Journal: Metabolites

Published: 2023

Background

Type 2 diabetes mellitus (T2DM) is associated with cardiovascular diseases, cognitive impairment, and dementia. There are notable sex differences in T2DM presentation and complications, with men diagnosed earlier and women facing higher mortality and complication rates. This study investigates the effects of T2DM on the brain metabolome, examining whole brain tissue from male and female db/db mice compared to wild-type controls. The research aims to identify sex-specific metabolic changes linked to T2DM and cognitive decline, filling a gap in understanding how sex influences brain metabolism in diabetes.

Materials & Methods

1) Animals

The study complied with humane care policies, with daily monitoring of food and water intake. Male and female db/db (T2DM model) and WT mice were obtained at 10 weeks and housed individually on an AIN-93M diet. At 17 weeks, glucose tolerance tests (GTT) were conducted, and euthanasia occurred at 18 weeks, with blood and brain tissue collected.

2) Peripheral Metabolic Assessment

Fasting serum glucose and glucose tolerance were assessed after an 8-hour fast. Blood samples were taken before and after glucose injection, and serum was collected via ventricular puncture for insulin and cholesterol analysis.

3) Metabolomics Tissue Preparation

Whole brain tissue from six mice per group was snap-frozen and powdered. For untargeted metabolomic analysis, tissue was homogenized in 80% methanol, followed by sonication and centrifugation.

4) LC-MS Analysis

Liquid chromatography-mass spectrometry (LC-MS) utilized an ACQUITY UPLC and Q Exactive MS. The mobile phase included formic acid and acetonitrile with a specific gradient. Peak areas were normalized using the total ion count method, with quality control samples included.

5) Pathway Analysis

Pathway overrepresentation analysis of altered metabolites was performed using IMPaLA software with KEGG identifiers.

6) Statistical Analysis

Statistical analyses for body weight, GTT, and serum parameters were conducted using Prism software, assessing normality and performing appropriate comparisons (p < 0.05). Metabolomics data were analyzed in MetaboAnalyst for clustering and comparisons.

Results

Confirmation of db/db Model

Both male and female db/db mice exhibited significantly higher body weight, fasting insulin, glucose, and cholesterol levels compared to WT mice (p < 0.05). The area under the curve (AUC) for a 2-hour glucose tolerance test (GTT) was also elevated in db/db mice.

Brain Metabolome Profiles

A total of 1,021 metabolites were identified. PLS-DA revealed distinct metabolic profiles between db/db and WT mice, with significant segregation regardless of sex. Heatmap analysis confirmed marked differences in metabolite patterns between groups.

PLS-DA comparisons of brain metabolites in response to T2DM in male and female mice.

Heatmap of group averages with hierarchical clustering of brain metabolites in response to T2DM in male and female mice.

T2DM-Induced Metabolite Changes

Pairwise comparisons showed T2DM altered 103 metabolites in male db/db mice and 65 in females (adjusted p < 0.05). In males, 67 metabolites increased (19 > 2-fold), while 36 decreased (14 < 0.5-fold). In females, 40 metabolites increased (7 > 2-fold) and 25 decreased (5 < 0.5-fold). A total of 80 metabolites were male-specific, 42 female-specific, and 23 were common, with 6 decreased and 17 increased in both sexes.

Effects of T2DM on the brain metabolome. Volcano plot of metabolites in T2DM db/db versus control WT for male (A) and female (B) mice.

Comparison of the brain metabolic response to T2DM in male and female mice. (A) Venn diagram comparison of significant metabolites in T2DM db/db vs. control WT male and female mice.

Metabolite Classes and Pathways

Fatty acyls comprised the largest category of altered metabolites (25–26%), followed by glycerophospholipids in males and a more balanced distribution among organoheterocyclic compounds, benzenoids, and organic acids in females (Figure 5A). Pathway analysis in males identified 25 significantly overrepresented pathways, primarily related to lipid metabolism, while no significant pathways were found in females.

Classification and pathway overrepresentation analysis of brain metabolites altered by T2DM. (A) Classifications of metabolites significantly altered by T2DM (db/db versus WT) in male and female mice. (B) All significant (Q < 0.05) pathways for metabolites altered by T2DM (db/db versus WT) in male mice (y-axis), and number of metabolites in each pathway (x-axis).

Impact of Sex on Metabolome

Significant differences were observed between male and female db/db mice (54 metabolites) and WT mice (15 metabolites). In db/db mice, more metabolites were lower in females (32 vs. 22 higher). Only three metabolites differed by sex in both genotypes.

Metabolite Classes and Pathways Altered by Sex

In db/db mice, fatty acyls (28%) and glycerophospholipids (22%) were predominant among sex-differentiated metabolites, while in WT mice, benzenoids (33%) were most common. Overrepresented pathways in db/db mice included lipid metabolism and GPCR signaling, with no significant pathways identified in WT mice.

Reference

1. Norman, Jennifer E., et al. "Sex modifies the impact of type 2 diabetes mellitus on the murine whole brain metabolome." Metabolites 13.9 (2023): 1012.