Background: Development of cost-effective platforms for identification of biomarkers is of paramount importance in low-income settings. The present work focuses on the development of a microfluidic paper-based analytical device (μPADs) for the diagnosis of osteoporosis by measuring three important bone biomarkers - calcium, alkaline phosphatase and vitamin D. Methods: Antibody-based detection is realized in μPAD reservoirs based on principle of colorimetric sensing, where μPAD images are captured using a smartphone. The results are compared with the calibration curves to give proper quantification of analytes. Conclusion: This device can be potentially developed into a point-of-care diagnostic device for low-income settings. To the best of our understanding, a μPAD platform for simultaneous diagnosis of multiple osteoporosis markers is not reported yet. © 2018 Newlands Press.