Azithromycin Analyzed with HPLC ELSD – AppNote – MicroSolv Technology Corporation

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ARTICLES, APPNOTES, FAQ (9187)
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June 6, 2022
Tanya

AppNotes Cogent Bidentate C8 / AppNotes Food and Beverages / AppNotes Library / ARTICLES, APPNOTES, FAQ

Retention of Macrolide Antibiotic

Azithromycin has weak UV absorbance and typical asymmetric peak profile with low Column efficiency in many HPLC-UV methods. This ELSD Method shows good retention and peak shape along with excellent sensitivity. This method is very reproducible with %RSD values less than 0.1%, as shown in the 10 injection overlay below.

Peak:
Azithromycin

Method Conditions
Column: Cogent Bidentate C8™, 4µm, 100Å
Catalog No.: 40008-10P
Dimensions: 4.6mm x 100mm
Mobile Phase:
A: Isopropanol
B: Acetonitrile / 0.1% Triethylamine (TEA) (v/v)
Gradient:

Time (minutes)	%B
0	100
1	100
2	85
3	85
4	100
5	100

Flow rate: 1.0 mL/minute
Detection: ELSD (Evaporative Light Scattering Detector) Gain: 9; Temperature: 80°C;
Injection vol.: 1μL
Sample Preparation: Reference standards (1 mg/mL) in diluent of 50:50 Acetonitrile / DI Water (v/v)
t₀: 1.50 Minutes
K’: 2

Note: Azithromycin is a semi-synthetic macrolide Antibiotic of the Azalide class. Azithromycin inhibits bacterial protein synthesis by binding to the 50S ribosomal subunit of the bacterial 70S ribosome.

Note 2: Capacity is determined using the following equation: k = (t_R-t_O)/t_O

t_R = Retention Time of an Analyte Peak
t_O = Retention Time of non-Retained Peak

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