Evaluation of total plasma homocysteine in Indian newborns using heel-prick samples
A Radha Rama Devi1, S M Naushad, Krishna C Prasad
1Diagnostic Division, Center for DNA Fingerprinting and Diagnostics, Hyderabad, India.
Insights
A modified High-Performance Liquid Chromatography (HPLC) method accurately analyzes total plasma homocysteine in newborn heel-prick samples. This study found the prevalence of hyperhomocysteinemia in Indian newborns to be 8.6%.
Area of Science:
- Biochemistry
- Neonatal Medicine
- Analytical Chemistry
Background:
- Elevated homocysteine levels, or hyperhomocysteinemia, are linked to adverse health outcomes.
- Accurate measurement of total plasma homocysteine is crucial for early detection and intervention.
- Existing methods may not be suitable for small-volume neonatal samples.
Purpose of the Study:
- To adapt and validate a High-Performance Liquid Chromatography (HPLC) method for quantifying total plasma homocysteine in neonatal heel-prick blood samples.
- To determine the prevalence of hyperhomocysteinemia among exclusively breast-fed Indian newborns.
Main Methods:
- Modification of a SBD-F based High-Performance Liquid Chromatography (HPLC) method with fluorescence detection.
- Validation using Bland and Altman analysis to ensure agreement with the original method.
- Analysis of plasma homocysteine levels in 607 randomly selected Indian newborn heel-prick samples.
Main Results:
- The modified HPLC method demonstrated high correlation (0.994) and acceptable limits of agreement with the original method for heel-prick samples.
- The mean plasma homocysteine level in Indian newborns was 6.99 (95% CI: 6.48-7.49), with no significant gender difference.
- Elevated homocysteine levels were detected in 8.6% of the studied Indian newborns.
Conclusions:
- The modified HPLC method is validated and suitable for analyzing total plasma homocysteine from small-volume newborn heel-prick samples.
- The prevalence of hyperhomocysteinemia in exclusively breast-fed Indian newborns is estimated at 8.6%.
- This validated method facilitates large-scale screening for hyperhomocysteinemia in neonatal populations.
Objective:
To estimate total plasma homocysteine levels in Indian newborns by modifying the existing SBD-F based High performance liquid chromotography (HPLC) method in order to enable analysis in newborn heel-prick samples and assess the prevalence of hyperhomocysteinemia in Indian newborns who are exclusively breast-fed.
Methods:
Reverse-phase HPLC with fluorescence detection for plasma homocysteine estimation and statistical analysis using student t-test.
Results:
SBD-F based HPLC method was modified and Bland and Altman analysis was carried out to assess agreement between original and modified methods. The correlation co-efficient was 0.994. The limits of agreement (-5.9, 6.3) were small enough to apply new method in place of the old for heel-prick sample analysis. Total plasma homocysteine analysis was carried out on heel-prick samples of 607 randomly selected newborns (331 males and 276 females). The mean plasma homocysteine estimated by this method in Indian newborns was 6.99 (95% CI: 6.48-7.49) with no appreciable gender effect (P=0.74). Elevated homocysteine levels were observed in 31 males and 21 females.
Conclusions:
Modified HPLC method is validated and can be used for homocysteine analysis on newborn heel-prick samples. Using this method, the prevalence of hyperhomocysteinemia in Indian newborns is 8.6%.
