A method for the measurement of histamine release by basophils using condensation to o-phthalaldehyde (OPT) and high performance liquid chromatography (HPLC) was described. Protein was precipitated from dextran sedimented and lysed peripheral leukocytes, and the histamine extracted into butanol, back into hydrochloric-acid, and then condensed to OPT. Fluorescence was measured on a filter fluorometer or by HPLC with a cyano-propyl column. A single fluorescent peak was seen with authentic OPT condensed histamine at 2.5 minutes. Chromatographic detection of histamine standards from 10 nanograms/milliliter to 1mg/ml was linear. Nonstimulated leukocyte supernatant extracts, perchloric-acid lysed leukocyte extracts and extracts from antibody stimulated leukocytes demonstrated single peaks that eluted at the same time as the authentic histamine. Unextracted lysed leukocyte preparations had decreased histamine peaks due to negative interferences that were removed by prior extraction of the histamine. Similar interference problems were seen using human urine extracts. The addition of interfering substances without extraction before derivatization resulted in interference of OPT/histamine fluorescence when measured with a fluorometer, but some positive interference could be filtered out by using a sharp cutoff filter.
We take your privacy seriously. You can review and change the way we collect information below.
These cookies allow us to count visits and traffic sources so we can measure and improve the performance of our site. They help us to know which pages are the most and least popular and see how visitors move around the site. All information these cookies collect is aggregated and therefore anonymous. If you do not allow these cookies we will not know when you have visited our site, and will not be able to monitor its performance.
Cookies used to make website functionality more relevant to you. These cookies perform functions like remembering presentation options or choices and, in some cases, delivery of web content that based on self-identified area of interests.
Cookies used to track the effectiveness of CDC public health campaigns through clickthrough data.
Cookies used to enable you to share pages and content that you find interesting on CDC.gov through third party social networking and other websites. These cookies may also be used for advertising purposes by these third parties.