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Application Note: (6R,?7R)?-3-?(Chloromethyl)?-?8-?oxo-?7-?[[2-?(2-?thienyl)?acetyl]?amino]?-5-?thia-?1-?azabicyclo[4.2.0]?oct-?2-?ene-?2-?carboxylic Acid (4-?Methoxyphenyl)?methyl Ester is an intermediate in synthesizing Nitrocefin (N493815), a chromogenic beta-lactamase substrate that undergoes colour change from yellow to red as the amide bond in the beta-Lactam ring is hydrolyzed by beta-lactamase. Nitrocefin undergoes colour changes induced by lactamases produced by Gram-positive and Gram-negative bacteria. Several studies have utilized the colour changing properties of Nitrocefin for the detection of beta-lactamase activity from bacterial cell extracts by isoelectric focusing and spectroscopy. Nitrocefin has also been used in studies involving beta-lactamase resistant antibiotics.