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UHPLC Analysis of Taxanes Using A Titan C18 Column and UV Detection

Taxanes are diterpenes produced by the plants of the genus Taxus (yews), and are widely used as chemotherapy agents. Taxane agents include paclitaxel (Taxol) and docetaxel (Taxotere). The role of taxanes in the treatment of breast cancer is becoming increasingly important. In clinical practice, the taxanes are now standard therapy in both early-stage and metastatic breast cancer. Taxanes are amont the most promising antitumor agents available today, of increasing importance given that cancer is now one of the major public health problems which needs to be dealt with urgently for the benefit of affected patients. Several ongoing experimental and clinical trials have supported the fact that even with their side effects and poor solubilities, taxanes are still the first lines of treatment chosen for breast, ovary, lung, and other metastatic cancers.

Furthermore, Taxanes are being explored for Alzheimer's treatment as noted in the recent Cortice Biosciences press release that states, in part, that they have begun a Phase 1 open-label dose-escalation trial designed to evaluate safety, tolerability and preliminary efficacy of the company's lead drug candidate, TPI 287, for treatment of patients with mild-to-moderate Alzheimer's disease*. TPI 287 is an abeotaxane, a taxoid that binds to and stabilizes microtubules (MTs) similarly to the taxanes, paclitaxel and docetaxel. A distinct advantage of TPI 287 over the taxanes is its ability to readily cross the blood-brain barrier*.

A challenge for formulation is that taxane medicines are poorly soluble in water. Therefore, there is quite a bit of research not only in taxane lead discovery and optimization but also in formulation. Intravenous injection is the most common delivery method although various drug delivery approaches explored so far include cosolvents, emulsions, micelles, liposomes, microspheres nanoparticles, cyclodextrins, pastes, and implants.

Taxanes are best analyzed by reversed phase liquid chromatography. Ultra High Performance Liquid Chromatography (UHPLC) provides the speed and sensitivity needed in drug discovery and clinical studies. Unfortunately, most UHPLC columns are twice as expensive as HPLC columns leading to increased study costs. Titan UHPLC columns are the outcome of the patented Ecoporous process. The Ecoporous process was developed to deliver UHPLC grade silica at a significant reduction in cost. The Titan columns' reduction in the cost to perform UHPLC will not only improve the productivity of pharmaceutical and contract research organizations currently employing UHPLC for medicinal chemistry, toxicology, and clinical manufacturing, but will also extend the opportunity for budget-conscious academic labs focused on translational research to achieve industry-standard analytical performance and throughput.

Shown in Figure 1 is the chromatographic analysis of six taxanes separated on a Titan C18 UHPLC column. The detection of the taxanes is accomplished using UV at 227 nm with complete separation in about 5 minutes. Excellent peak shape and efficiency are demonstrated in this data set. Titan columns are manufactured with monodisperse 1.9 micron silica that yield the same performance benefits as core-shell type columns.

Figure 1. UHPLC Analysis of Taxanes

Figure 1. UHPLC Analysis of Taxanes

Method Conditions:
column: Titan C18 10 cm x 2.1 mm, 1.9 µm (Product No. 577124-U)
mobile phase: (A) water; (B) (50:50, methanol:acetonitrile)
gradient: 60 to 80% B in 4.5 min; 80% B held for 1.5 min
flow rate: 0.4 mL/min
pressure: 6090 psi (420 bar)
column temp.: 40 °C
detector: UV, 227 nm
injection: 2 µL
sample: 25 µg/mL in 70:30, water:methanol


In conclusion, Titan UHPLC columns offer a tremendous value in terms of performance and cost for both research and applied disciplines. Titan UHPLC columns should help expedite important drug discovery and clinical research projects in anti-tumor and oncology as well as other pharmaceutical disciplines.

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Materials

     

 References

  1. Cortice Announces Enrollment of a Clinical Trial Targeting Tau Dysfunction in Mild-to-Moderate Alzheimer's Disease. Jan. 21, 2014, http://globenewswire.com/news-release/2014/01/21/603701/10064872/en/Cortice-Announces-Enrollment-of-a-Clinical-Trial-Targeting-Tau-Dysfunction-in-Mild-to-Moderate- Alzheimer-s-Disease.html?parent=619922 , (accessed Apr 2014)
  2. Fitzgerald, D.P.; Emerson, D.L.; Qian, Y.; Anwar, T.; Liewehr, D.J.; Steinberg, S.M.; Silberman, S.; Palmieri, D.; Steeg, P.S. TPI_287, a new taxane family member, reduces the brain mestastatic colonization of breast cancer cells. Mol. Cancer Ther. 2012, 11, 1959.

 

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