Benefits of Mass Detection Using the ACQUITY QDa Mass Detector for Routine Botanical Authentication

Significance of the Topic

Botanical ingredients are widely used in dietary supplements, herbal medicines, cosmetics, and personal care products. However, plant misidentification and adulteration present safety and quality challenges. Reliable, routine authentication methods help ensure standardization, consumer safety, and regulatory compliance.

Objectives and Study Overview

This study assesses the feasibility of using the Waters ACQUITY QDa Mass Detector, coupled with UPLC, for routine authentication of North American black cohosh (Actaea racemosa). A blind evaluation of commercial and laboratory-prepared samples gauges method accuracy and compares mass detection with evaporative light scattering detection (ELSD).

Methodology and Used Instrumentation

A targeted UPLC-MS workflow was developed to detect characteristic marker compounds in black cohosh extracts. Key steps included:

• Sample preparation: Extraction and dilution in 70% methanol, followed by 0.2 µm filtration.
• Chromatography: ACQUITY UPLC H-Class system equipped with an ACQUITY BEH C18 (1.7 µm, 2.1×100 mm) column at 50 °C; 9 min gradient elution (water/formic acid and acetonitrile/methanol/formic acid).
• Mass detection: ACQUITY QDa Mass Detector in ESI+ mode (scan range 200–1000 Da), optimized at 1.5 kV capillary voltage, 10 V cone voltage, probe temperature 300 °C, 5 Hz sampling, with a diverter valve to waste before 0.8 min and after 9 min.
• Data processing: Empower 3 CDS software used to build a custom MS library, extract ion chromatograms (XIC), and implement an automated authenticity protocol based on retention time, peak area, library match, and relative ion intensity.

ELSD conditions (2424 ELS) were recorded for comparative evaluation but were found less specific.

Main Results and Discussion

• Method optimization: MS parameters were tuned to maximize molecular ion intensities, with cone voltage optimization for cimiracemoside C.
• Marker selection: Among four candidate markers (cimifugin, cimiracemoside C, actein, 27-deoxyactein), cimiracemoside C (m/z 621 Da, RT 5.8 min) exhibited the most distinct fingerprint in authentic samples.
• Automated authentication protocol: A decision tree in Empower 3 evaluated presence in RT window, peak area thresholds, MS library match, and ion abundance criteria, enabling rapid classification.
• Sample analysis: All four commercial samples and two laboratory-contaminated mixtures (5% and 10% adulteration) were correctly classified as authentic or inauthentic.
• Comparison with ELSD: MS XICs provided clear differentiation of authentic black cohosh, while ELSD chromatograms lacked sufficient specificity to distinguish genuine and adulterated samples.

Benefits and Practical Applications

• The ACQUITY QDa is an accessible, cost-effective MS detector suitable for routine QC laboratories.
• High selectivity of mass detection simplifies analysis of complex botanical matrices.
• Empower 3–driven automation reduces operator expertise requirements and standardizes decision making.
• Enhanced confidence in botanical authenticity supports product safety, quality assurance, and regulatory adherence.

Future Trends and Applications

Future developments may involve expanding MS libraries to cover a broader range of botanicals, integrating chemometric and AI-based pattern recognition, and combining with high-resolution MS for trace adulterant detection. Continued miniaturization and cost reduction of MS detectors will further democratize routine botanical authentication across the nutraceutical and personal care industries.

Conclusion

The combination of UPLC with the ACQUITY QDa Mass Detector and Empower 3 software provides a robust, user-friendly, and affordable solution for routine botanical authentication. Targeted marker detection and automated data workflows enable rapid, accurate classification of black cohosh samples, outperforming traditional ELSD approaches and supporting high standards of safety and quality.

References

1. Gafner S. Laboratory guidance documents, ABC-AHP-NCNPR botanical adulterants program. 2015.
2. Geng P, Harnly JM, Sun J, Zhang M, Chen P. Feruloyl dopamine-O-hexosides are efficient marker compounds as orthogonal validation for authentication of black cohosh (Actaea racemosa) – a UHPLC-HRAM-MS chemometrics study. Anal Bioanal Chem. 2017 Apr;409(10):2591–2600.
3. He K, Pauli GF, Zheng B, Wang H, Bai N, Peng T, Roller M, Zheng Q. Cimicifuga species identification by high-performance liquid chromatography-photodiode array/mass spectrometric/evaporative light scattering detection for quality control of black cohosh products. J Chromatogr A. 2006 Apr 21;1112(1–2):241–254.
4. Sharaf M, Yuk J, Yu K, Wrona M, Isaac G. Chemical profiling of Actaea species and commercial products using UPLC-QTof-MS. Poster presented at the 9th Joint Natural Products Conference, July 24–27, 2016; Copenhagen, Denmark.
5. Masada S. Authentication of the botanical origin of Western herbal products using Cimicifuga and Vitex products as examples. J Nat Med. 2016;70:361–375.
6. Avula B, Wang Y-H, Smillie TJ, Khan IA. Quantitative determination of triterpenoids and formononetin in rhizomes of black cohosh (Actaea racemosa) and dietary supplements by using UPLC-UV/ELS detection and identification by UPLC-MS. Planta Med. 2009;75:381–386.
7. Jiang B, Kronenberg F, Nuntanakorn P, Qiu M-H, Kennelly EJ. Evaluation of botanical authenticity and phytochemical profile of black cohosh products by high-performance liquid chromatography with selected ion monitoring liquid chromatography–mass spectrometry. J Agric Food Chem. 2006 May 3;54(9):3242–3253.
8. Whiting PW, Clouston A, Kerlin P. Black cohosh and other herbal remedies associated with acute hepatitis. Med J Aust. 2002;177:432–435.
9. Yang J, Rainville P. Automated 2-Dimensional Marker’s Fingerprint Analysis for Routine Botanical Authentication Using the ACQUITY QDa Mass Detector. Waters Application Note. 2018 May.
10. United States Pharmacopeia-National Formulary (USP 40–NF 35). United States Pharmacopeial Convention; 2017.