초록 ▼

Nitrosyl (FeII) hemoglobin can be detected in biological samples, using a method which involves injections of samples into a photolysis cell, prior to detection of chemiluminescence generated by the reaction between nitric oxide and ozone. This method is useful for monitoring the levels of nitric ox

Nitrosyl (FeII) hemoglobin can be detected in biological samples, using a method which involves injections of samples into a photolysis cell, prior to detection of chemiluminescence generated by the reaction between nitric oxide and ozone. This method is useful for monitoring the levels of nitric oxide bioactivity in both normal physiological states, and disease states, such as septic shock, atherosclerosis, thrombosis, hyperhomocysteinemia, pulmonary hypertension, malignancy, infections and central nervous system disorders.

대표청구항 ▼

[ What is claimed is:] [8.] A method for assaying thiol-bound NO (nitric oxide) in S-nitrosohemoglobin in a sample of purified hemoglobin, comprising the steps of:a) dividing the sample of purified hemoglobin containing said S-nitrosohmoglobin into a first aliquot and a second aliquot;b) contacting

[ What is claimed is:] [8.] A method for assaying thiol-bound NO (nitric oxide) in S-nitrosohemoglobin in a sample of purified hemoglobin, comprising the steps of:a) dividing the sample of purified hemoglobin containing said S-nitrosohmoglobin into a first aliquot and a second aliquot;b) contacting said first aliquot with mercury ions in excess over protein concentration, in the purified hemoglobin to obtain a mercury-treated aliquot;c) isolating from said mercury-treated aliquot a mercury-treated low molecular weight fraction, and from said second aliquot a second low molecular weight fraction;d) contacting said mercury-treated low molecular weight fraction with excess thiol under acidic conditions and contacting said second low molecular weight fraction with excess thiol under acidic conditions, thereby producing S-nitrosothiol in said mercury-treated low molecular weight fraction and in said second low molecular weight fraction;e) subjecting said mercury treated low molecular weight fraction and said second low molecular weight fraction to photolysis thereby liberating nitric oxide from said S-nitrosothiol in said low molecular weight fractions;f) reacting said liberated nitric oxide in said mercury-treated low molecular weight fraction and in said second low molecular weight fraction with ozone, thereby generating chemiluminescent signals;g) measuring said chemiluminescent signals, thereby obtaining a measurement of thiol-bound nitric oxide in said mercury-treated low molecular weight fraction and a measurement of thiol-bound NO in said second low molecular weight fraction; andh) determining said thiol-bound nitric oxide in S-nitrosohemoglobin in said sample of purified hemoglobin from a difference between said measurement of said thiol-bound nitric oxide in said mercury-treated low molecular weight fraction and said measurement of said thiol-bound nitric oxide in said second low molecular weight fraction.