Analysis of Geniposidic Acid and Chlorogenic Acid in Tochu Tea Using a Triple Quadrupole LC/MS/MS [LCMS-8030]

Importance of the Topic

Tochu tea, derived from the leaves of Eucommia ulmoides, is a traditional remedy valued for its health benefits. Key bioactive compounds include geniposidic acid, known for blood pressure reduction, and chlorogenic acid, a major polyphenol. Accurate quantification of these constituents is critical for quality control, standardization of herbal products and pharmacological research.

Objectives and Overview of the Study

This study aimed to develop and validate a sensitive and selective LC MS MS method using a triple quadrupole instrument to quantify geniposidic acid and chlorogenic acid in Tochu tea. The approach focused on optimizing sample preparation, chromatographic separation and MRM detection conditions to achieve reliable results in complex plant extracts.

Methodology and Instrumentation

Sample preparation involved steeping 20 g of Tochu leaves in boiling water for three 10-minute cycles, adjusting the extract to 1 L, diluting 1 in 1000 and filtering through a 0.2 µm membrane. Chromatographic separation was carried out on an ODS column with a gradient of formic acid in water and acetonitrile containing formic acid. Detection employed electrospray ionization in negative mode and MRM transitions of m/z 373>123 for geniposidic acid and m/z 353>191 for chlorogenic acid. Collison energies and quadrupole bias voltages were optimized to maximize signal intensity and specificity.

Used Instrumentation

• LC MS MS system: Shimadzu LCMS-8030 triple quadrupole
• Column: Shim-pack VP-ODS, 150 mm × 2.0 mm i.d., 5 µm
• Mobile phases: A 0.1 % formic acid in water, B acetonitrile with 0.1 % formic acid
• Gradient: 10 % B to 20 % B over 10 min, then 10 % B until 20 min
• Flow rate: 0.2 mL/min; injection 2 µL
• Ionization: ESI negative, probe voltage –3.5 kV, DL temperature 300 °C, block heater 500 °C, drying gas 20 L/min

Main Results and Discussion

MS MS spectra of standard solutions confirmed major product ions at m/z 123 for geniposidic acid and m/z 191 for chlorogenic acid. MRM chromatograms demonstrated clear separation and consistent retention times. Calibration curves showed excellent linearity over 5 to 10000 ppb for geniposidic acid and 5 to 5000 ppb for chlorogenic acid, with correlation coefficients above 0.999. Analysis of the diluted Tochu tea extract yielded concentrations of 600–700 ppb for both analytes, corresponding to approximately 600–700 ppm in the undiluted infusion. No significant matrix interference was observed.

Benefits and Practical Applications of the Method

• High sensitivity and selectivity for trace quantification in complex matrices
• Wide dynamic range supports quality control across diverse product concentrations
• Rapid analysis suitable for routine screening in research and industrial laboratories
• Scalable approach that can be adapted to other plant-derived compounds

Future Trends and Potential Applications

Advances may include coupling with high-resolution MS for untargeted profiling, microflow LC for reduced sample consumption, and automation for high-throughput screening. Integration with metabolomics platforms could expand the analysis to broader phytochemical networks. Green extraction techniques and portable MS devices may enable in-field testing of herbal teas.

Conclusion

The presented triple quadrupole LC MS MS method provides a robust, accurate and efficient tool for simultaneous quantification of geniposidic acid and chlorogenic acid in Tochu tea. Its performance in terms of sensitivity, linearity and matrix tolerance makes it valuable for quality assurance and research on herbal medicines.

References

• Shimadzu Corporation. Analysis of Geniposidic Acid and Chlorogenic Acid in Tochu Tea Using a Triple Quadrupole LC MS MS. Application News No. C82; February 2012.