HPLC-Knowledge Compact

„HPLC-Knowledge compact“: Essential Topics condensed taught

In these half-day workshops (8.30-12.00 am and 1.00-4.30 pm) we highlight a specific HPLC-topic very intensive (group work, exercises, question-answer-games). The workshops are suited perfectly for experienced HPLC-users. Different workshops can of course be arbitrary combined, however some topics fit very well together, e.g. in the morning Comparison and selection of HPLC-RP-columns“ and in the afternoon „Which gradient should I use?” or in the morning „Intelligent method development“ and in the afternoon How to prove robustness“ etc. Participants can in principle attend only the morning-workshop or only the afternoon-workshop as well or of course both.

Development of robust HPLC-methods – „that´s” the way!

  • Method development under time pressure
  • Proven/successful combinations of column/mobile phase/pH-value
  • Tests to prove column stability – basic requirements for a robust routine method

Tips und hints how to prove the peak-homogeneity

  • Quick tests in absence of LC-MS-coupling
  • What is „best“? DAD, LC/MS/MS, less injection volume or a polar pre-column?
  • Tests for the optimum “benefit-cost-ratio” – the “5-min method”

Tips for successful method transfers

  • Typical organisational and analytical errors during method transfer
  • „Do´s und dont´s“-list to avoid future trouble
  • Review of real cases

“Testing“/verification of robustness in HPLC

  • What is crucial but has been often „forgotten“ during testing robustness?
  • Hints how to realize lack of robustness by carefully reading the method description
  • Schemes and plans to test robustness dependent to the characteristics of the method and the matrix

Correct integration and correct evaluation of chromatographic data

  • Poor resolution, „snippy“ peaks close to LOQ and/or drifting baseline – how to integrate correct, e. g: drop, „valey to valey“, tangential or exponential skim?
  • Which chromatographic parameters and which software settings affect how strong peak area, peak shape and LOD/LOQ?
  • What does mean a „too“ small RSD and what should be the consequence? Is this value a usual deviation or an outlier?

Choosing the „right“ eluent – incl. gradient elution (mobile phase in detail plus a little bit gradient)

  • How to quickly find the appropriate composition of the mobile phase and of the right pH-value dependent on the analyte or the real aim of the method?
  • When methanol, when acetonitril, when, which pH-value (and buffer)?
  • Exercises for optimization and troubleshooting of gradient runs

HPLC in pharmaceutical companies; specific requirements und tips for the daily work

  • System Suitability Test, SST; what, how, how often – requirements und reality
  • Which guidelines are clear and which are “interpretable”? News and trends by the FDA, USP, ICH, Pharm.Eur.
  • Which claims are for the authorities very important, where lies the emphasis by the tests?

Such a way you get the optimal output from your HPLC/UHPLC

  • HPLC vs. UHPLC: Benefits and drawbacks in the daily work
  • Why do some modern columns in modern equipments sow a bad performance? Typical hardware-pitfalls
  • Pro´s and Con´s of binary and ternary pumps, pre-heater or heat exchanger capillary etc. What are really the major/particular differences of modern HPLC/UHPLC-equipment?

Typical errors during sample separation

  • Lost of analyte during the sample preparation procedure – the main reasons
  • Kind of diluent and solving handling  – their influence on peak shape, number of peaks and the reproducibility of the peak area
  • How to avoid errors by preparing the mobile phase

Your HPLC-method(s) in the focus of your in-house-training!

We prepare a custom-made workshop in which your methods are in focus – up to 4 methods per ½ -day workshop. Considering your whishes in the training we focus on sample preparation (method A), or/and optimization (method B), or/and integration (method C)…We support you, so that method becomes more robust, this one faster and in the third case you can save a lot of mobile phase by changing just few things.

The progress of the workshop is really individual, this particular training is based on your aim, e. g:

  • You´d like to come to optimized methods – incl. hand-on in your lab at your HPLC-equipment? We work with you in your lab!
  • You´d like to convert a 30-40 min. method to a in 5-10 min robust method with no or litlle revalidation work? Do you like to use your UHPLC really as „UHPLC“?
  • You´d like to learn how to find an alternative, more robust column for these particular compounds? We bring a numerous of columns to try them out with you in your lab.

Find the best training concept for you