--- title: "Prohibited Substances in Racehorse Urine: Validated SPE Method with Clean Screen® DAU" description: Discover a validated method for screening 63 prohibited substances in racehorse urine, ensuring compliance and integrity in the equine doping control process. image: https://blog.chromatographydirect.com/hubfs/Canva%20images/600%20x%20308%20CD%20Blog%20UCT%20-%20DAU%20cartridges.png --- [Skip to content](https://blog.chromatographydirect.com/prohibited-substances-in-racehorse-urine#main-content) [![cd-logo-header](https://blog.chromatographydirect.com/hs-fs/hubfs/Logos/cd-logo-header.png?width=180&height=68&name=cd-logo-header.png)](https://www.chromatographydirect.com/) - [Products](https://www.chromatographydirect.com/products/vials-and-caps) Show submenu for Products - [Vials & Caps](https://www.chromatographydirect.com/products/vials-and-caps) - [Sample Preparation](https://info.chromatographydirect.com/uct-sample-preparation-products-chromatography-direct) - [Filtration Products](https://www.chromatographydirect.com/products/filtration-products) - [LC Columns & Accessories](https://www.chromatographydirect.com/products/lc-columns-and-accessories) - [Instrument Spares](https://www.chromatographydirect.com/products/instrument-spares) - [Plates & Mats](https://www.chromatographydirect.com/products/plates-and-mats) - [Flash Chromatography](https://www.chromatographydirect.com/products/flash-chromatography) - [Solvent Safety](https://www.chromatographydirect.com/products/solvent-safety) - [GC Columns & Accessories](https://www.chromatographydirect.com/products/gc-columns-and-accessories) - [Dissolution](https://www.chromatographydirect.com/products/dissolution) - [Clinical Kits, Standards and Controls](https://www.chromatographydirect.com/products/clinical-kits-standards-and-controls) - [New Products](https://www.chromatographydirect.com/new-products) - [Knowledge Base](https://www.chromatographydirect.com/knowledge-base) - [About Us](https://www.chromatographydirect.com/about-us) - [Offers](https://www.chromatographydirect.com/offers) - [Blog](https://blog.chromatographydirect.com) Open main navigation Close main navigation - [Products](https://www.chromatographydirect.com/products/vials-and-caps) Show submenu for Products - [Vials & Caps](https://www.chromatographydirect.com/products/vials-and-caps) - [Sample Preparation](https://info.chromatographydirect.com/uct-sample-preparation-products-chromatography-direct) - [Filtration Products](https://www.chromatographydirect.com/products/filtration-products) - [LC Columns & Accessories](https://www.chromatographydirect.com/products/lc-columns-and-accessories) - [Instrument Spares](https://www.chromatographydirect.com/products/instrument-spares) - [Plates & Mats](https://www.chromatographydirect.com/products/plates-and-mats) - [Flash Chromatography](https://www.chromatographydirect.com/products/flash-chromatography) - [Solvent Safety](https://www.chromatographydirect.com/products/solvent-safety) - [GC Columns & Accessories](https://www.chromatographydirect.com/products/gc-columns-and-accessories) - [Dissolution](https://www.chromatographydirect.com/products/dissolution) - [Clinical Kits, Standards and Controls](https://www.chromatographydirect.com/products/clinical-kits-standards-and-controls) - [New Products](https://www.chromatographydirect.com/new-products) - [Knowledge Base](https://www.chromatographydirect.com/knowledge-base) - [About Us](https://www.chromatographydirect.com/about-us) - [Offers](https://www.chromatographydirect.com/offers) - [Blog](https://blog.chromatographydirect.com) - [Contact us](https://info.chromatographydirect.com/contact-chromatography-direct) [Contact us](https://info.chromatographydirect.com/contact-chromatography-direct) --- Aug 17, 2026, 9:30:03 AM # Prohibited Substances in Racehorse Urine: Validated SPE Method with Clean Screen® DAU [Chromatography Direct](https://blog.chromatographydirect.com/author/chromatography-direct) Share: [facebook-f icon](http://www.facebook.com/share.php?u=https://blog.chromatographydirect.com/prohibited-substances-in-racehorse-urine) [linkedin-in icon](http://www.linkedin.com/shareArticle?mini=true&url=https://blog.chromatographydirect.com/prohibited-substances-in-racehorse-urine) [twitter icon](https://twitter.com/intent/tweet?url=https://blog.chromatographydirect.com/prohibited-substances-in-racehorse-urine) [envelope icon](mailto:?body=https://blog.chromatographydirect.com/prohibited-substances-in-racehorse-urine) ## The Application Horse racing is a multi-million pound industry with zero tolerance for doping. The International Federation of Horseracing Authorities (IFHA) maintains a list of prohibited substances, and antidoping labs need methods that screen broadly across drug classes from a single extraction. That means one SPE cartridge handling anesthetics, analgesics, sedatives, opioids, beta-blockers, stimulants, anti-inflammatories, and more, all from one of the dirtiest biological matrices in routine testing. A Turkish antidoping laboratory validated exactly that. ![600 x 308 CD Blog UCT - DAU cartridges](https://blog.chromatographydirect.com/hs-fs/hubfs/Canva%20images/600%20x%20308%20CD%20Blog%20UCT%20-%20DAU%20cartridges.png?width=600&height=310&name=600%20x%20308%20CD%20Blog%20UCT%20-%20DAU%20cartridges.png) ## The Method **63 prohibited substances** extracted from racehorse urine and analysed by LC-MS/MS, validated to 2021/808/EC guidelines. **Product used:** Clean Screen® DAU, 500 mg / 3 mL (P/N CSDAU503) ### Sample Preparation 1. Mix 14 mL urine with internal standard, 1 M phosphate buffer (pH 5.8), ammonium sulfate, protease, and β-glucuronidase 2. Incubate 1.5 hours at 55°C (hydrolysis) 3. Transfer 7 mL, centrifuge, dilute with phosphate buffer and water 4. Adjust to pH 7 with 10% NaOH ### SPE Protocol (Automated, Gilson Aspect GX-274) 1. **Condition** with water, methanol, and phosphate buffer (0.1 M, pH 6) 2. **Load** sample 3. **Wash** with 1 M acetic acid (dry), then methanol (dry) 4. **Elute** with ethyl acetate:aqueous ammonia (97:3 v/v) 5. **Evaporate** and reconstitute for LC-MS/MS ### The Clogging Problem (and Two Solutions) Horse urine is viscous and protein-rich. During development, cartridge clogging became an issue. The researchers solved it by adding protease during hydrolysis and precipitating proteins with ammonium sulfate. The alternative: **UCT's XtrackT® DAU columns**. Large-particle silica (80–200 µm) delivers better, more consistent flow through dirty samples without requiring additional sample pre-treatment. If your lab processes high volumes of equine urine, XtrackT® eliminates the clogging problem at the sorbent level. ## Performance - Screening limits and low LODs achieved for all 63 substances - Matrix effects: -17.88% to +19.7% (effective matrix removal confirmed) - No interferences detected in selectivity/specificity testing - Full compliance with 2021/808/EC validation criteria ## Why Chromatography Direct The home of chromatography consumables. We stock both Clean Screen® DAU and XtrackT® DAU for equine antidoping work. High-quality consumables at unrivalled prices, making your lab budget go further. XtrackT® DAU has previously been validated for 89 banned or controlled racing industry drugs in horse urine. Whether you need standard or large-particle SPE, we carry the range and we price it for labs running high-throughput antidoping screening. Specialists, not generalists. --- **Reference** Kabil, E.; Göktaş, E. F. Development and Validation of Confirmatory Quantitative and Screening Methods for 63 Prohibited Substances in Horse Urine Using LC-MS/MS. *Chromatographia* 2025, 88, 151–168. DOI: [10.1007/s10337-024-04384-2](https://doi.org/10.1007/s10337-024-04384-2) [clean screen](https://blog.chromatographydirect.com/tag/clean-screen), [XtrackT](https://blog.chromatographydirect.com/tag/xtrackt), [CSDAU](https://blog.chromatographydirect.com/tag/csdau), [DAU](https://blog.chromatographydirect.com/tag/dau) --- [![cd-logo-header](https://blog.chromatographydirect.com/hs-fs/hubfs/Logos/cd-logo-header.png?width=170&height=64&name=cd-logo-header.png "cd-logo-header")](https://www.chromatographydirect.com/) © Copyright Chromatography Direct Ltd 2026 [Privacy Policy](https://www.chromatographydirect.com/privacy-policy) [Cookie Policy](https://www.chromatographydirect.com/cookie-policy) [Terms of Use](https://www.chromatographydirect.com/terms-of-use) [Terms & Conditions](https://www.chromatographydirect.com/terms-and-conditions) [Trademarks](https://www.chromatographydirect.com/trademarks) **Chromatography Direct Ltd** 11 Regent Park, Booth Drive, Park Farm Industrial Estate, Wellingborough, Northants, NN8 6GR United Kingdom **T** [+44 (0) 1925 599070](tel:+441925599070) **F** [+44 (0) 1925 599071](tel:+441925599071) **E** [sales@chromatographydirect.com](mailto:sales@chromatographydirect.com) facebook-f icon linkedin-in icon twitter icon ```json { "@context" : "https://schema.org", "@type" : "BlogPosting", "author" : { "@type" : "Person", "name" : "Chromatography Direct", "url" : "https://blog.chromatographydirect.com/author/chromatography-direct" }, "dateModified" : "2026-08-17T08:30:03.683Z", "datePublished" : "2026-08-17T08:30:03.000Z", "headline" : "Prohibited Substances in Racehorse Urine: Validated SPE Method with Clean Screen® DAU", "image" : [ "https://blog.chromatographydirect.com/hubfs/Canva%20images/600%20x%20308%20CD%20Blog%20UCT%20-%20DAU%20cartridges.png" ], "mainEntityOfPage" : { "@id" : "https://blog.chromatographydirect.com/prohibited-substances-in-racehorse-urine", "@type" : "WebPage" }, "publisher" : { "@type" : "Organization", "logo" : { "@type" : "ImageObject", "url" : "https://blog.chromatographydirect.com/hubfs/chrom-direct-logo-large-1.jpg" }, "name" : "Chromatography Direct Ltd" } } ```