ISSN

0974-4150 (Online)
0974-4169 (Print)


Author(s): Shrimant N Panaskar, Dinesh Yadav, Susheel K Singh, Mahesh H Hampe, Akshata Shivalkar

Email(s): shrimant.p@preventine.com

DOI: 10.5958/0974-4150.2019.00069.5   

Address: Shrimant N Panaskar1*, Dinesh Yadav2, Susheel K Singh3, Mahesh H Hampe4, Ms. Akshata Shivalkar5
1MSc PhD, Senior Scientist, Preventi Ne Life Care, Navi Mumbai, Maharashtra (India).
2MSc, Quality Manager, Preventi Ne Life Care, Navi Mumbai, Maharashtra (India)
3B. Tech-PGDM, CEO, Preventi Ne Life Care, Navi Mumbai, Maharashtra (India)
4MD, DNB, Consultant Biochemist, Preventi Ne Life Care, Navi Mumbai, Maharashtra (India)
*Corresponding Author

Published In:   Volume - 12,      Issue - 6,     Year - 2019


ABSTRACT:
Objective: To develop a simple and reliable method for quantification of biotinidase (EC.3.5.1.12) activity in dried blood spot and serum by a modification of existing colorimetric screening test. Method: The assay involves incubation of DBS, serum sample and controls in 30 µL phosphate buffer containing synthetic substrate B-PABA, EDTA and 10µL of 40mM disulfide reducing reagent Mercaptoethanol at 37ºC for 4 hours. The reaction was further stopped by TCA addition for hemoglobin denaturation. After the coupling reaction was started, the absorbance was measured by a microplate-reader. Results: A present rapid method using Mercaptoethanol (ME) for biotinidase assay from dried blood spots was developed, validate and compare with Yamaguchi et al method for dried blood spot (n=360). The method performance with DBS was compare with serum by using Wolf et al method. The correlation coefficients obtained by plotting the values obtained by modified method against the values by Wolf et and Yamaguchi et al methods were 0.86 and 0.96 respectively. Conclusion: Biotinidase activities in DBS (n360) and in serum (n=50) were always enhanced by 2-mercaptoethanol (ME). The modified method has reduced the processing and incubation time up to 4 Hr. and it is significantly correlate with reference method. The present method applicable for newborn screening in DBS as well as test for biotinides deficiency in serum. Modified method is very cost effective due to less run time, precision and accuracy.


Cite this article:
Shrimant N Panaskar, Dinesh Yadav, Susheel K Singh, Mahesh H Hampe, Akshata Shivalkar. Enhancement of Biotinidase activity in dried blood spot by Disulfide Reducing Reagent and Comparative Evaluation with Reference Method. Asian J. Research Chem. 2019; 12(6):366-371. doi: 10.5958/0974-4150.2019.00069.5


Recomonded Articles:

Author(s): Vatchavai Bhaskara Raju, Bonthu Mohan Gandhi, Kamatham Srinivas Sumanth, Kolli Srinivas, Tupakula N Venkata Lakshmi Neeraja

DOI: 10.5958/0974-4150.2017.00030.X         Access: Open Access Read More

Author(s): VB Kshirsagar, UA Deokate, VB Bharkad, SS Khadabadi

DOI:         Access: Open Access Read More

Author(s): Ravi Pratap Pulla, B.S. Sastry, Y. Rajendra Prasad, N. Appala Raju

DOI:         Access: Open Access Read More

Author(s): Tentu Nageswara Rao, Botsa Parvatamma, Karri Apparao, Jakka Mastan

DOI: 10.5958/0974-4150.2017.00059.1         Access: Open Access Read More

Author(s): P Nagaraju, K Krishnachaithanya, VDN Srinivas, SVN Padma

DOI:         Access: Open Access Read More

Author(s): K. Balamuralikrishna, B. Syamasundar

DOI:         Access: Open Access Read More

Author(s): Gopalasatheeskumar K, Ariharasiva Kumar G, Sengottuvel T, Sanish Devan V, Srividhya V

DOI: 10.5958/0974-4150.2019.00062.2         Access: Closed Access Read More

Author(s): P Kalaiselvi, R Vijay Amirtharaj, T Venkatachalam, N Senthil Kumar

DOI:         Access: Open Access Read More

Author(s): R. Xavier Arulappa, M.Sundarapandian, Rithesh Singh, S.Venkataraman

DOI:         Access: Open Access Read More

Author(s): Krishna Veni Nagappan, Meyyanathan S N, Rajinikanth B Raja, Elango Kannan

DOI:         Access: Open Access Read More

Author(s): Mitesh D Phale, Purnima D Hamrapurkar

DOI:         Access: Open Access Read More

Author(s): V Kiran Kumar, N Appala Raju, Namratha Rani, JVLN Seshagiri Rao, T Satyanarayana

DOI:         Access: Open Access Read More

Author(s): Cijo John, Mohd Idris, P. Sudhakar, Priyankar Ghosh, K.M. Varshney, S.K. Shukla

DOI:         Access: Open Access Read More

Author(s): Zarna Dedania, Ronak Dedania, G Vidhyasagar, Bhavna Patel, Chetan Ramolia, Vaishali Karkhanis

DOI:         Access: Open Access Read More

Author(s): C.K.Oza, Ranju Nijhawan, M. K. Pandya, A. J. Vyas, A. I. Patel

DOI:         Access: Open Access Read More

Asian Journal of Research in Chemistry (AJRC) is an international, peer-reviewed journal devoted to pure and applied chemistry..... Read more >>>

RNI: Not Available                     
DOI: 10.5958/0974-4150 

Popular Articles


Recent Articles




Tags