Salvianolic Acid B Analysis Service

What is Salvianolic Acid B?

Salvianolic Acid B (Sal B) is a major bioactive polyphenolic compound extracted primarily from Salvia miltiorrhiza (Danshen), a cornerstone of Traditional Chinese Medicine. Structurally characterized by multiple phenolic hydroxyl groups, Sal B exhibits potent antioxidant, anti-inflammatory, cardioprotective, and neuroprotective properties. It plays a critical role in vascular protection, platelet aggregation inhibition, and modulation of cellular signaling pathways such as MAPK and TGF-β. Due to its high polarity and complex molecular structure (C₃₆H₃₀O₁₆, MW: 718.61 g/mol), precise quantification and metabolomic mapping of Sal B remain a scientific priority in drug discovery, herbal pharmacology, and functional food research.

Why Analyze Salvianolic Acid B?

Enabling Pharmacological Validation and Process Optimization

Precise Salvianolic acid B analysis is essential for:

• Pharmacokinetics & Bioavailability: Monitoring absorption, distribution, metabolism, and excretion in preclinical or clinical models.
• Herbal Product Standardization: Ensuring batch-to-batch consistency in botanical formulations and dietary supplements.
• Metabolite Profiling: Characterizing in vivo or in vitro biotransformation pathways of Sal B.
• Stability Testing: Assessing compound degradation under various processing or storage conditions.
• Therapeutic Targeting: Supporting mechanism-of-action studies and pathway-level insights.

Creative Proteomics delivers robust and reproducible Salvianolic Acid B analysis services that meet regulatory and scientific requirements for high-impact research and development.

Salvianolic Acid B Analysis Service Offered by Creative Proteomics

• Quantitative Analysis – Accurate measurement of Salvianolic Acid B concentrations in plasma, tissue, urine, and plant extracts using LC-MS/MS.
• Metabolite Profiling – Identification and quantification of Sal B-related metabolites including oxidative, sulfated, and glucuronidated forms.
• Pharmacokinetics Study – Time-point analysis of Sal B to evaluate absorption, distribution, metabolism, and excretion (ADME).
• Stability Testing – Assessment of Sal B stability and degradation under conditions such as heat, light, pH, and long-term storage.
• Metabolic Pathway Analysis – Mapping of biosynthetic and downstream metabolic pathways using integrated MS and bioinformatics.
• Tissue and Biofluid Analysis – Detection of Sal B in blood, urine, liver, kidney, brain, and other biological matrices.
• Plant and Herbal Extract Testing – Comprehensive profiling of Sal B in raw herbs, extracts, powders, and formulated products.
• Toxic Metabolite Screening – Detection of potentially toxic or reactive metabolites for safety and risk assessment.
• Precursor and Derivative Identification – Analysis of structurally related compounds such as Danshensu, caffeic acid, and other salvianolic acids.

List of Detected Salvianolic Acid B and Related Metabolites

Advantages of Salvianolic Acid B Assay

• Ultra-Sensitive Detection: Detection limit as low as 5 pg/mL using Agilent 6495C Triple Quadrupole LC/MS, suitable for trace-level quantification in complex matrices.
• Wide Dynamic Range: Linear quantification across ≥4 orders of magnitude, enabling accurate measurement from low to high concentrations without repeated dilutions.
• High Precision: Intra- and inter-batch variability <10% RSD, ensuring consistent and reproducible results across multiple runs.
• Excellent Recovery: Optimized extraction yields >95% recovery in plasma, tissue, urine, and plant matrices.
• Comprehensive Target Panel: Simultaneous quantification of 15+ related compounds, including Danshensu, caffeic acid, and salvianolic acids A–E.
• Fast Turnaround: Standard reports delivered in 14–28 business days; expedited services available.
• Validated Quantification: All assays performed with ≥6-point calibration curves, achieving R² ≥ 0.995 for quantitative accuracy.

Workflow for Salvianolic Acid B Analysis Service

Technology Platform for Salvianolic Acid B Analysis Service

High-Performance Liquid Chromatography (HPLC): Agilent 1260 Infinity II HPLC

Triple Quadrupole Mass Spectrometry: Agilent 6495C Triple Quadrupole LC/MS

Agilent 6495C Triple Quadrupole (Figure from Agilent)

Agilent 1260 Infinity II HPLC (Fig from Agilent)

Sample Requirements for Salvianolic Acid B Analysis Service

FAQ of Salvianolic Acid B Analysis Service

Learn about other Q&A.

Load More FAQs

Salvianolic Acid B Analysis Service Case Study

Title: Simultaneous Quantification of Aspirin, Its Metabolite Salicylic Acid, and Salvianolic Acid B in Human Plasma Using UPLC-MS/MS

Journal: International Journal of Analytical Chemistry

Published: 2021

• Background
• Methods
• Results
• Conclusion
• Reference

Salvianolate injection, derived from danshen (a widely used Chinese medicinal herb), is frequently combined with aspirin (ASA) in clinical practice to treat coronary heart disease, particularly in patients with stable angina. Salvianolic acid B (Sal B) is the major active component of salvianolate injection and has demonstrated cardiovascular protective effects, including anti-inflammatory and anti-platelet properties.

Previous studies suggest potential drug-drug interactions (DDIs) between ASA and danshen, which may influence their pharmacokinetic profiles and therapeutic outcomes. However, there has been no existing bioanalytical method capable of simultaneously quantifying ASA, its metabolite salicylic acid (SA), and Sal B in human plasma due to differences in their chemical and physical properties.

To address this gap, the authors developed and validated a rapid, sensitive UPLC-MS/MS method for the simultaneous determination of ASA, SA, and Sal B. This method enables accurate pharmacokinetic analysis in clinical DDI studies and supports the safe and rational combined use of salvianolate injection and aspirin in cardiovascular therapy.

• Sample Preparation: Human plasma samples processed via liquid-liquid extraction with ethyl acetate, dried under nitrogen, and reconstituted before injection.
• Chromatography: Separation on a BEH C18 column using gradient elution (0.5% formic acid in water and acetonitrile), 0.4 mL/min flow rate, 2 μL injection volume.
• Detection: Negative ESI mode with MRM transitions for Sal B, ASA, SA, and internal standard (IS).
• Linearity & Sensitivity: Calibration curves (r > 0.99); LLOQ at 5 ng/mL.
• Validation Parameters: Method validated for specificity, accuracy, precision, matrix effect, recovery, and stability per regulatory standards.
• Clinical Application: Successfully applied in a 16-subject PK-DDI clinical trial to evaluate interactions between salvianolate injection and aspirin.

UPLC-MS/MS Optimization

• Negative ion mode selected for better signal intensity.
• Optimized MRM transitions:
  • Sal B: m/z 717.2→519.1
  • ASA: m/z 179.1→137.0
  • SA: m/z 136.8→65.0
  • IS: m/z 321.0→152.0
• Chromatographic separation resolved ASA-SA overlap (resolution > 1.5).
• Peak shapes improved by lowering mobile phase pH (0.5% formic acid outperformed 0.1%).

Sample Preparation

• Liquid-liquid extraction chosen over protein precipitation to ensure sensitivity, especially for low ASA levels.
• Final reconstitution solvent: MeOH/ACN/H₂O (40:40:20, v/v/v) optimized for linear calibration curves of all analytes.

Extraction Recovery & Matrix Effect

• High recoveries: Sal B: ~85%, ASA: ~95%, SA: ~94%.
• Minimal matrix effect after IS normalization (RSD < 4.6%).

Precision & Accuracy

• Intra-/inter-day accuracy within ±11%; precision <10% for all analytes—compliant with bioanalytical standards.

Stability

• Analytes stable for 4 h at room temp (with acidifier), 60 days at −80°C, and 24 h in autosampler at 4°C.
• Acidification required to prevent ASA degradation to SA during plasma handling.

Clinical Application

• Applied to plasma samples from 16 subjects.
• Most Sal B and SA levels quantifiable; ASA often below LLOQ due to fast metabolism.
• PK data showed reduced drug exposure and altered absorption/elimination in the aspirin + salvianolate group, indicating a drug-drug interaction (DDI).

Representative MRM chromatograms of Sal B ASA, SA, and IS in plasma. (a) Blank plasma. (b) LLOQ sample (Sal B ASA, and SA at 5 ng/mL; IS at 20 ng/mL). (c) Plasma sample 4 h after administration.

Table 1: MS/MS ion acquisition parameters of Sal B, ASA, SA, and IS

Table 2: Calibration curves and correlation coefficients of Sal B, ASA, and SA.

Developed a robust, sensitive, and validated UPLC-MS/MS method for simultaneous quantification of Sal B, ASA, and SA in human plasma. Method is suitable for clinical pharmacokinetic studies, especially DDI assessments involving traditional Chinese medicine and Western drugs. Provides a reference for future in vivo analysis of similar compound combinations in integrated medicine research.

Reference

1. Cao, Weiyi, et al. "Simultaneous Quantification of Aspirin, Its Metabolite Salicylic Acid, and Salvianolic Acid B in Human Plasma Using UPLC‐MS/MS." International Journal of Analytical Chemistry 2021.1 (2021): 6620868. https://doi.org/10.1155/2021/6620868