Biogenic amines were analyzed in the Vanderbilt University Neurochemistry Core as described previously8 (link). Briefly, biogenic amine concentrations were determined utilizing an Antec Decade II (oxidation: 0.65) electrochemical detector operated at 33 °C. 20μl samples of the supernatant were injected using a Water 2707 autosampler onto a Phenomenex Kintex C18 HPLC column (100 × 4.60 mm, 2.6 μm). Biogenic amines are eluted with a mobile phase consisting of 89.5% 0.1 M TCA, 10-2 M sodium acetate, 10-4 M EDTA and 10.5 % methanol (pH 3.8). Solvent was delivered at 0.6 ml/min using a Waters 515 HPLC pump. Using this HPLC solvent the following biogenic amines were eluted in the following order: Norepinephrine, Epinephrine, DOPAC, Dopamine, 5-HIAA, HVA, 5-HT, and 3-MT. Data acquisition was managed by Empower software (Waters Corporation, Milford, MA USA). Isoproterenol (5 ng/mL) was included in the homogenization buffer for use as an internal standard to quantify the biogenic amines of interest.
Liu Z., Yang N., Dong J., Tian W., Chang L., Ma J., Guo J., Tan J., Dong A., He K., Zhou J., Cinar R., Wu J., Salinas A.G., Sun L., Kumar M., Sullivan B.T., Oldham B.B., Pitz V., Makarious M.B., Ding J., Kung J., Xie C., Hawes S.L., Wang L., Wang T., Chan P., Zhang Z., Le W., Chen S., Lovinger D.M., Blauwendraat C., Singleton A.B., Cui G., Li Y., Cai H, & Tang B. (2022). Deficiency in endocannabinoid synthase DAGLB contributes to early onset Parkinsonism and murine nigral dopaminergic neuron dysfunction. Nature Communications, 13, 3490.
Other organizations :
National Institute on Aging, National Institutes of Health, Capital Medical University, Xiangya Hospital Central South University, Central South University, Peking University, National Institute of Environmental Health Sciences, National Institute on Alcohol Abuse and Alcoholism, Huazhong University of Science and Technology, Union Hospital, Dalian Medical University, First Affiliated Hospital of Dalian Medical University, Shanghai Jiao Tong University, Ruijin Hospital, Center for Life Sciences
Concentrations of biogenic amines: Norepinephrine, Epinephrine, DOPAC, Dopamine, 5-HIAA, HVA, 5-HT, and 3-MT
control variables
Temperature of electrochemical detector set at 33 °C
Sample injection volume of 20 μl
Mobile phase composition: 89.5% 0.1 M TCA, 10-2 M sodium acetate, 10-4 M EDTA and 10.5% methanol (pH 3.8)
Mobile phase flow rate of 0.6 ml/min
Use of Isoproterenol (5 ng/mL) as an internal standard in the homogenization buffer
positive controls
None explicitly mentioned
negative controls
None explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
