Lab Med Online.  2023 Jan;13(1):13-21. 10.47429/lmo.2023.13.1.13.

The negative role of Vitamin C in total cholesterol and triglycerides tests

• Ko YK
• Lee HS
• Lee K
• Song J
• Park HD

• KMID: 2552720
• DOI: http://doi.org/10.47429/lmo.2023.13.1.13

Abstract

Background
This study aimed to determine the cause of low total cholesterol and triglyceride levels based on the Trinder reaction by reviewing cases of negative interference.
Methods
We analyzed the results of total cholesterol and triglyceride tests performed over 19 months. Sixty-three test results from 48 patients showing abnormally low total cholesterol or triglyceride levels were included. Lipid profile (total cholesterol, triglyceride, HDL-cholesterol, and LDL-cholesterol) was evaluated with Roche Diagnostics (Germany) and Sekisui Medical (Japan) reagents by using Cobas c702 analyzer (Roche). In vitro lipid profile tests were also determined using both above reagents for serum spiked with nine different concentrations of vitamin C.
Results
Total cholesterol and triglyceride levels measured using Roche reagents (without ascorbate oxidase) were significantly lower than those using Sekisui reagents containing ascorbate oxidase respectively. HDL-cholesterol and LDL-cholesterol levels showed no significant difference between the two company reagents. Malignancy was diagnosed in 83% of the patients, and the patients have the possibility of receiving high-dose vitamin C therapy. In addition, 27% of the patients were diagnosed with chronic kidney disease. At the level of 25.0 mg/dL vitamin C to the serum, total cholesterol and triglyceride levels, measured using Roche reagents, reduced by 27% and 95%, respectively, but were rarely reduced when using Sekisui reagents.
Conclusions
When negative interference is suspected in the total cholesterol or triglyceride of patients, the specimens need to be retested with the reagent containing ascorbate oxidase, and the patient’s medical record should be reviewed thoroughly.

Keyword

Ascorbic acid; Ascorbate oxidase; Analytic interference; Cholesterol; Enzymatic assays; Triglycerides

Figure

• 

  Fig. 1 Flow-chart of the study outline.

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  Fig. 2 Summary of lipid profile measurements between the two reagents. Box plots show the 25th percentile, median (horizontal lines inside the boxes), and the 75th percentile of the data values of total cholesterol, triglyceride, HDL-cholesterol, and LDL-cholesterol measured with Roche reagents (gray-colored box plot) and Sekisui reagents (uncolored box plot). Crossbar lines on box plot whiskers show the 5th and 95th percentile of data values. The “ * “ mark above the box plot indicates the reagents containing ascorbate oxidase.

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  Fig. 3 Difference between plotted results obtained from Roche and Sekisui reagents. (A) Total cholesterol (CHOL); (B) Triglyceride (TG); (C) HDL-cholesterol (HDLC); (D) LDL-cholesterol (LDLC). The solid line and gray boxed area represent the absolute mean difference and its 95% CI, respectively. The dotted line represents the absolute mean difference of 0. The total cholesterol and triglyceride results using Roche and Sekisui reagents showed a large difference. The HDL-cholesterol and LDL-cholesterol results of Roche and Sekisui reagents showed no significant differences.

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  Fig. 4 Effect of concentration of ascorbic acid on interference in lipid profile testing between the two reagents. Measurement of pooled serum values of (A) total cholesterol (Roche: filled circle with solid line, Sekisui: filled circle with dotted line), (B) triglyceride (Roche: filled triangle with solid line, Sekisui: filled triangle with dotted line), (C) HDL-cholesterol (Roche: empty circle with solid line, Sekisui: empty circle with dotted line), and (D) LDL-cholesterol (Roche: empty triangle with solid line, Sekisui: empty triangle with dotted line) are converted to the percentage of reduction. Concentrations of triglyceride with Roche TRIGL and total cholesterol with Roche CHOL2 were reduced by more than 90% after adding 25 mg/dL and 87.5 mg/dL of ascorbic acid, respectively. (Reagents with ascorbate oxidase: Pureauto S CHO-N, Pureauto S TG-N, HDL3, LDL3. Reagents without ascorbate oxidase: CHOL2, TRIGL, Cholestest N HDL, Cholestest N LDL).

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