Pregnenolone 17α,21,21,21-d4 | CAS 61574-54-7

OVERVIEW  of  Pregnenolone 17α,21,21,21-d4 | CAS 61574-54-7

 

Pregnenolone 17α,21,21,21-d4 | CAS 61574-54-7 is a stable isotope-labeled analogue of pregnenolone, a key steroidal precursor involved in the biosynthesis of glucocorticoids, mineralocorticoids, progesterone, and androgens. As a central intermediate in the steroidogenesis pathway, pregnenolone plays an essential role in endocrine research, adrenal function evaluation, and steroid metabolism studies. The incorporation of four deuterium atoms at the 17α and 21 positions provides a distinctive mass shift without altering the biochemical or structural behavior of the native molecule.

This deuterated standard is engineered for high-precision LC-MS/MS and GC-MS quantification of pregnenolone in biological matrices, including serum, plasma, urine, tissue extracts, and clinical samples. Due to its stable isotopic incorporation and near-identical physicochemical properties to pregnenolone, Pregnenolone-d4 is ideal for use as an internal standard in validated steroid hormone assays where accurate and reproducible quantification is critical.

---

CHEMICAL INFORMATION

• Chemical Name: Pregnenolone 17α,21,21,21-d4

• CAS Number: 61574-54-7

• Molecular Formula: C21H28D4O2

• Isotopic Enrichment: ≥ 98% D

• Chemical Class: Steroidal ketone; deuterated endogenous steroid

• Molecular Weight: 320.51 g/mol

The selective deuteration at metabolically relevant positions ensures analytical separation from native pregnenolone while maintaining identical chromatographic retention and extraction efficiency.

---

STRUCTURAL & ANALYTICAL PROPERTIES

Pregnenolone-d4 closely mirrors the physicochemical characteristics of the unlabeled steroid:

• Excellent solubility in organic solvents such as methanol, acetonitrile, and dichloromethane

• Chemical stability suitable for long-term storage and routine analytical workflows

• Consistent ionization behavior in both positive and negative electrospray modes

• Retention time parity with native pregnenolone under reversed-phase LC conditions

• Predictable fragmentation patterns due to minimal isotopic influence on backbone structure

The +4 Da isotopic shift is sufficient to create clean MS separation without affecting the molecule’s biological or physicochemical function, making it a robust internal standard for highly sensitive assays.

---

APPLICATIONS  of  Pregnenolone 17α,21,21,21-d4 | CAS 61574-54-7

 

Steroid Hormone Quantification (LC-MS/MS)

Pregnenolone 17α,21,21,21-d4 is extensively used as an internal standard for the quantification of pregnenolone in:

• Clinical hormone testing

• Endocrine and adrenal function studies

• Reproductive research

• Neurosteroid quantitation

• Stress biomarker evaluation

Its stable isotopic signature ensures precise, reproducible quantitation across diverse biological matrices.

Steroidogenesis Pathway Analysis

Pregnenolone is the first hormonal product of cholesterol conversion via CYP11A1. The d4-labeled version supports:

• Mapping of early steroid biosynthesis

• Monitoring conversion to progesterone and other downstream steroids

• Evaluating mitochondrial steroidogenic activity

• Studying disruptions caused by disease, pharmaceuticals, or environmental factors

Pharmacokinetics & Metabolic Profiling

In drug development and metabolism research, the compound enables:

• Tracking pregnenolone turnover

• Identifying metabolic intermediates

• Assessing enzymatic activity, particularly 3β-HSD and CYP family enzymes

• Understanding steroid clearance and transformation pathways

Its structural fidelity ensures matched extraction recovery and ionization compared to the native analyte.

Clinical, Diagnostic & Research Method Validation

The use of a deuterated internal standard is essential for meeting validation criteria such as:

• Accuracy and precision

• Linearity

• Matrix effect compensation

• Stability studies

• Carryover and reproducibility assessments

Pregnenolone-d4 is fully compatible with both clinical diagnostic workflows and high-throughput research laboratories.

---

ADVANTAGES OF USING PREGNENOLONE-D4

• Distinct +4 Da mass difference for clear MS resolution

• Minimal effect on retention, fragmentation, or extraction

• Highly reliable correction for matrix suppression/enhancement

• Supports FDA/EMA bioanalytical method validation guidelines

• Essential for accurate steroid hormone quantification in complex matrices

• Enhances confidence in clinical and research steroid profiling

---

CONCLUSION

Pregnenolone 17α,21,21,21-d4 (CAS 61574-54-7) is a high-quality, stable isotope-labeled internal standard essential for modern steroid hormone analysis. Its exceptional isotopic purity, chemical stability, and close structural alignment with native pregnenolone make it suitable for LC-MS/MS quantification, steroidogenesis studies, metabolic profiling, and regulated clinical bioanalysis. This deuterated analogue ensures precision, reproducibility, and reduced analytical variability, making it a critical reference material for endocrinology, pharmacology, and biomedical research.

 

Learn more about,

Deuterated Polymers: A Cornerstone Guide to Synthesis, Applications, and Future Trends

Availability of All the Deuterated Chemicals at ResolveMass Laboratories Inc. 

ResolveMass Laboratories: Leading Deuterated Chemical Synthesis Company in the United States. 

Deuterated Internal Standards for LC-MS: Selection & Custom Synthesis 

How to Choose the Right Deuterated Labelled Chemical Synthesis Company in Canada 

How to Choose the Right Deuterium Labelled Compounds Supplier for Your Lab 

Deuterium-Labelled Compounds — Synthesis, Applications & Ordering 

Custom Synthesis of Deuterated Chemicals