August/September 2011


43


Using Online Sample Preparation with LC-MS/MS to Speed Up the Process of Analysing Pharmaceutical and Personal Care Products


Kevin McHale, Mark Sanders Thermo Fisher Scientific Somerset, NJ *Address for correspondence kevin.mchale@thermofisher.com


There is growing environmental concern regarding the health impact of trace levels of pharmaceuticals and personal care products (PPCPs) in water resources. Although the term PPCP was only recently coined, these bioactive chemicals have been in existence for decades, and their effect on the environment is now recognised as an important area of research. PPCPs include prescription and over- the-counter therapeutic drugs, cosmetics, and nutraceuticals such as vitamins. These substances can find their way into water through human activity, residues from pharmaceutical manufacturing and residues from hospitals. Some PPCPs are easily broken down and processed by the human body or can degrade quickly in the environment. Others, however, are not broken down so easily and enter domestic sewers. Until recently individuals’ contribution to the combined chemical load in the environment has gone largely unnoticed.


In response to this concern, the U.S Environmental Protection Agency (EPA) recently published Method 1694, which determines dozens of PPCPs in water, soil, sediment and biosolids by high performance liquid chromatography combined with tandem mass spectrometry1


(HPLC-MS/MS).


The method, yet to be promulgated, uses solid phase extraction (SPE) of water samples followed by HPLC-MS/MS analysis using a single transition for each compound to achieve low nanogram/liter (ng/L) limits of quantitation (LOQs). The target analytes in the EPA method are divided into four groups, with each group representing one HPLC-MS/MS run. Three of the groups are extracted under acidic conditions; the fourth is extracted under basic conditions. These SPE methods can use up to 1 L of sample. Although not sample limited, the storage of large bottles of water requires a great deal of refrigeration space. In addition, manual SPE of 1 L of sample requires several hours of preparation.


One of the opportunities afforded in the analysis of PPCPs in water is to reduce the time required for sample preparation and analysis while maintaining the required sensitivity at the ng/L level and the selectivity to positively identify the analyte of interest. This article will describe a method for online sample preparation and analysis using the Thermo Scientific EQuan system. This method couples a fast HPLC system with two LC columns – one for pre-concentration of the sample and the second for the analytical analysis – and a high performance


Compound


Trimethoprim Cefotaxime Norfloxacin


Ofloxacin


Ciprofloxacin Lomefloxacin Enrofloxacin Sarafloxacin Flumequine Lincomycin Azithromycin Erythromycin Tylosin


Class Antibiotic Antibiotic, cephalosporin Compound


Antibiotic, fluoroquinolone Chlorotetracycline Antibiotic, fluoroquinolone Doxycycline


Antibiotic, fluoroquinolone Anhydrotetracycline Antibiotic, fluoroquinolone Carbamazepine Antibiotic, fluoroquinolone Fluoxetine Antibiotic, fluoroquinolone Miconazole Antibiotic, fluoroquinolone Thiabendazole Antibiotic, macrolide Antibiotic, macrolide Antibiotic, macrolide Antibiotic, macrolide


Diphenhydramine Acetaminophen Codeine


Anhydroerythromycin Antibiotic, macrolide Clarithromycin Roxithromycin Ampicillin


Antibiotic, macrolide Antibiotic, macrolide Antibiotic, penicillin


Penicillin G Penicillin V Oxacillin


Cloxacillin Metformin* Sulfadiazine Sulfathiazole Sulfamerazine Sulfamethazine Sulfamethizole


Antibiotic, penicillin Antibiotic, penicillin Antibiotic, penicillin Antibiotic, penicillin Antidiabetic


Antibiotic, sulfa Antibiotic, sulfa Antibiotic, sulfa Antibiotic, sulfa Antibiotic, sulfa


Sulfachloropyridazine Antibiotic, sulfa Sulfamethoxazole Sulfadimethoxine Minocycline


Antibiotic, sulfa Antibiotic, sulfa


Oxytetracycline 4-epi-Tetracycline Tetracycline


Antibiotic, tetracycline Antibiotic, tetracycline Antibiotic, tetracycline Antibiotic, tetracycline


*Metformin was analysed using HILIC **PPCPs not included in E.P.A 1694


Table 1. PPCPs analysed 1EPA Method 1694: Pharmaceuticals and personal care products in water, soil, sediment, and biosolids by HPLC/MS/MS, December 2007, EPA-821-R-08-002


Cimetidine Ranitine


Digoxigenin Digoxin


Diltiazem


Dextromethorphan** Atenolol


Metoprolol Propranolol Albuterol


Midazolam


1-OH Midazolam 1-OH Alprazolam Alprazolam


Nordiazepam


1,7-Dimethylxanthine Caffeine


Benzoylecgonine Cocaine


Cocaethylene Cotinine


Class


4-epi-Chlorotetracycline Antibiotic, tetracycline Demeclocycline


Antibiotic, tetracycline Antibiotic, tetracycline


Antibiotic, tetracycline Antibiotic, tetracycline Anticonvulsant Antidepressant Antifungal


Antihelmintic Antihistamine Analgesic


Analgesic, narcotic Antiacid reflux Antiacid reflux Antiarrythmic Antiarrythmic


Antiarrythmic, benzothiazepine Antitussive


Beta-blocker Beta-blocker Beta-blocker


Bronchodialator


Sedative, benzodiazepine Sedative, benzodiazepine Sedative, benzodiazepine Sedative, benzodiazepine Sedative, benzodiazepine Stimulent Stimulent Stimulent Stimulent Stimulent Stimulent


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