Hydrocodone with Acetaminophen Analyzed by HPLC - AppNote
May 17, 2012
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Simple, Robust Assay Method
Click HERE for Column Ordering Information.
Hydrocodone can yield poor Peak shapes in many conventional Reversed Phase C18 Methods due to its tertiary amine group. The USP Assay Method for Hydrocodone in combination with Acetaminophen uses Triethylamine as a Mobile Phase additive to improve the Peak shape.
In this Method however, only Trifluoroacetic Acid is needed in the Mobile Phase for a Symmetrical Hydrocodone Peak. In addition, the Repeatability of the analysis is excellent as the five-run overlay in the Figure shows. Retention time %RSDs of < 0.1% were obtained for both Peaks.
Column: Cogent Phenyl Hydride™, 4 µm, 100 Å
Catalog No.: 69020-7.5P
Dimensions: 4.6 x 75 mm
Mobile Phase:
-- A: DI Water / 0.1% Trifluoroacetic Acid (TFA)
-- B: Acetonitrile / 0.1% Trifluoroacetic Acid (TFA)
Gradient:
Temperature: 35˚C
Injection vol.: 5 µL
Flow rate: 1.0 mL / minute
Detection:
-- 0–4 minutes: UV @ 295 nm
-- 4–10 minutes: UV @ 210 nm
Sample Preparation: One tablet containing 5 mg Hydrocodone / 500 mg Acetaminophen was ground and diluted to 100 mL with 50:50 Solvent A / Solvent B mixture. The solution was then sonicated 10 minutes and filtered with a 0.45 µm Nylon Syringe Filter (MICROSOLV Tech Corp.).
t 0 : 0.9 minutes
Note: Hydrocodone is a semi-synthetic opioid used as a narcotic analgesic to relieve moderate to severe pain. The formulation which includes Acetaminophen is marketed under several trade names, including Vicodin® and Lortab®.
Attachment No 145 Hydrocodone with Acetaminophen Analyzed by HPLC pdf 0.5 Mb Download File
Click HERE for Column Ordering Information.
Hydrocodone can yield poor Peak shapes in many conventional Reversed Phase C18 Methods due to its tertiary amine group. The USP Assay Method for Hydrocodone in combination with Acetaminophen uses Triethylamine as a Mobile Phase additive to improve the Peak shape.
In this Method however, only Trifluoroacetic Acid is needed in the Mobile Phase for a Symmetrical Hydrocodone Peak. In addition, the Repeatability of the analysis is excellent as the five-run overlay in the Figure shows. Retention time %RSDs of < 0.1% were obtained for both Peaks.
Peaks:
1. Acetaminophen,
2. Hydrocodone
Column: Cogent Phenyl Hydride™, 4 µm, 100 Å
Catalog No.: 69020-7.5P
Dimensions: 4.6 x 75 mm
Mobile Phase:
-- A: DI Water / 0.1% Trifluoroacetic Acid (TFA)
-- B: Acetonitrile / 0.1% Trifluoroacetic Acid (TFA)
Gradient:
| Time (minutes) | %B |
|---|---|
| 0 | 5 |
| 1 | 5 |
| 5 | 60 |
| 6 | 5 |
Injection vol.: 5 µL
Flow rate: 1.0 mL / minute
Detection:
-- 0–4 minutes: UV @ 295 nm
-- 4–10 minutes: UV @ 210 nm
Sample Preparation: One tablet containing 5 mg Hydrocodone / 500 mg Acetaminophen was ground and diluted to 100 mL with 50:50 Solvent A / Solvent B mixture. The solution was then sonicated 10 minutes and filtered with a 0.45 µm Nylon Syringe Filter (MICROSOLV Tech Corp.).
t 0 : 0.9 minutes
Note: Hydrocodone is a semi-synthetic opioid used as a narcotic analgesic to relieve moderate to severe pain. The formulation which includes Acetaminophen is marketed under several trade names, including Vicodin® and Lortab®.
Attachment No 145 Hydrocodone with Acetaminophen Analyzed by HPLC pdf 0.5 Mb Download File