Use of Fructosyl Peptide Oxidase for HbA1c Assay

Yonehara, Satoshi; Inamura, Norio; Fukuda, Misao; Sugiyama, Koji

doi:10.1177/1932296815569573

Cited by 12 publications

(4 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Blood samples were collected into two sodium fluoridecontaining blood collection tubes for glucose measurement (Venoject II, Terumo Co., Tokyo, Japan). One was immediately centrifuged and subjected to HbA1c measurement in blood cells by EA (CinQ HbA1c, Arkray, Inc., Kyoto, Japan) [4]. The other sample in a test tube stand was transferred to Arkray Inc. by courier in a refrigerated state, and HPLC-HbA1c and EA-HbA1c were measured the day following blood sample collection.…”

Section: Laboratory Methodsmentioning

confidence: 99%

Sample transport and/or storage can cause falsely low HbA1c levels in blood cells measured by enzymatic assay

Koga¹,

Okumiya

Ishibashi

2019

Diabetol Int

View full text Add to dashboard Cite

Although HbA1c measurement by enzymatic assay (EA-HbA1c) is widely used in health-screening settings in Japan, recent studies have suggested lower EA-HbA1c levels as compared with HbA1c levels measured by high-performance liquid chromatography (HPLC-HbA1c). Hypothesizing that falsely low levels of EA-HbA1c are attributable to hemolysis caused by sample transport and/or storage, we measured EA-HbA1c in blood cells and whole blood after sample transport and compared them with HPLC-HbA1c levels. Blood samples were collected from ten non-diabetic individuals into sodium fluoride-containing blood collection tubes and immediately measured for EA-HbA1c in blood cells. After transport, the blood samples were again subjected to measurement of EA-HbA1c levels in blood cells and whole blood the following day. These EA-HbA1c levels were compared with HPLC-HbA1c levels. EA-HbA1c levels in blood cells measured immediately after sample collection did not significantly differ from HPLC-HbA1c levels. Transported blood samples showed hemolysis and significantly lower EA-HbA1c levels in blood cells, as compared with HPLC-HbA1c levels, whereas no significant difference was observed between EA-HbA1c levels in whole blood and HPLC-HbA1c levels. Transported blood samples showed hemolysis and falsely low EA-HbA1c levels in blood cells. Hemolysis caused by sample transport and/or storage might be responsible for the falsely low EA-HbA1c levels. This should be kept in mind, because falsely low HbA1c levels may lead to a false-negative diagnosis of diabetes.

show abstract

Section: Laboratory Methodsmentioning

confidence: 99%

Sample transport and/or storage can cause falsely low HbA1c levels in blood cells measured by enzymatic assay

Koga¹,

Okumiya

Ishibashi

2019

Diabetol Int

View full text Add to dashboard Cite

show abstract

“…Since the intact protein HbA 1c is not a substrate for the used FPOX, the fructosamine residue does not react with R1. However, during this incubation step, endogenous fructosamine derivatives present in the blood sample are oxidized and the released hydrogen peroxide is degraded by the HRP present, thereby increasing the specificity of HbA 1c detection later [47]. After addition of reagent 2 (R2), containing a protease and a leuco dye, HbA 1c is enzymatically cleaved by the protease.…”

Section: Enzymatic Oxidative Degradation Of Fructosamine Fru-val-his mentioning

confidence: 99%

Direct glucosone-based synthesis and HILIC-ESI-MS/MS characterization of N-terminal fructosylated valine and valylhistidine for validation of enzymatic HbA1c assays in the diagnosis of diabetes mellitus

Gerke

Buchholz

Müller³

et al. 2019

Anal Bioanal Chem

View full text Add to dashboard Cite

Naturally occurring fructosamines are of high clinical significance due to their potential use in diabetes mellitus monitoring (quantification of fructosylated hemoglobin, HbA 1c ) or for the investigation of their reactivity in consecutive reactions and harmfulness towards the organism. Here we report the specific synthesis of the fructosylated dipeptide L-valyl-L-histidine (Fru-Val-His) and fructosylated L-valine (Fru-Val). Both are basic tools for the development and validation of enzymatic HbA 1c assays. The two fructosamine derivatives were synthesized via a protected glucosone intermediate which was coupled to the primary amine of Val or Val-His, performing a reductive amination reaction. Overall yields starting from fructose were 36% and 34% for Fru-Val and Fru-Val-His, respectively. Both compounds were achieved in purities > 90%. A HILIC-ESI-MS/MS method was developed for routine analysis of the synthesized fructosamines, including starting materials and intermediates. The presented method provides a well-defined and efficient synthesis protocol with purification steps and characterization of the desired products. The functionality of the fructosylated dipeptide has been thoroughly tested in an enzymatic HbA 1c assay, showing its concentration-dependent oxidative degradation by fructosyl-peptide oxidases (FPOX).

show abstract

“…HbA1c adalah protein terglikasi di mana glukosa melekat pada terminal-N dari rantai β hemoglobin (Yonehara et al, 2015). Kadar HbA1c mewakili kontrol glikemik rata-rata selama 2-3 bulan terakhir (Ryden et al, 2014).…”

Section: Pendahuluanunclassified

Pengaruh Lama Penyimpanan 5 Jam dan 10 Jam pada Suhu 2-8 0C Terhadap Kadar Glycated Hemoglobin (HbA1c)

Prihandono

Waluyo

2019

JMK

View full text Add to dashboard Cite

ABSTRAKTes HbA1c digunakan untuk mendiagnosa Diabetes Mellitus. Pada tes laboratorium HbA1c kadar glukosa tidak dipengaruhi oleh fluktuasi glukosa harian. Maka dari itu tes HbA1c digunakan sebagai tes pengendalian Diabetes Mellitus. Tujuan penelitian ini adalah untuk membuktikan apakah ada pengaruh lama penyimpanan 5 jam dan 10 jam pada suhu 2-8 oC terhadap kadar HbA1c. Jenis penelitian adalah penelitian Observasi Analitik. Menggunakan 20 sampel darah whole blood yang dilakuan pemeriksaan kadar HbA1c. Sampel diberi perlakuan yaitu tanpa penyimpanan, penyimpanan pada suhu 2-8 oC selama 5 jam, penyimpanan pada suhu 2-8 0C selama 10 jam. Dari tes Kolmogorov Smirnov, menunjukkan bahwa data tidak terdistribusi normal dengan p value 0,025 (P <0,05). Untuk menguji apakah ada pengaruh lama penyimpanan 5 jam dan 10 jam pada suhu 2-8 oC terhadap kadar HbA1c, dilanjutkan dengan uji Kruskal Wallis. Uji Kruskal Wallis menunjukkan bahwa nilai signifikan 0,929 dengan demikian P > 0,05 dapat disimpulkan bahwa tidak ada pengaruh lama penyimpanan 5 jam dan 10 jam pada suhu 2-8 oC terhadap kadar HbA1c. Proses penundaan pemeriksaan dan penyimpanan sampel darah perlu diperhatikan, karena kesalahan pada faktor pra analitik akan mempengaruhi kondisi sampel whole blood yang akan dilakukan analisa HbA1c yang akan dapat memberikan hasil tinggi palsu atau rendah palsu. Kata kunci : Lama Penyimpanan 5 Jam Dan 10 Jam, Suhu 2-8 0C, HbA1c

show abstract

Use of Fructosyl Peptide Oxidase for HbA1c Assay

Cited by 12 publications

References 8 publications

Sample transport and/or storage can cause falsely low HbA1c levels in blood cells measured by enzymatic assay

Sample transport and/or storage can cause falsely low HbA1c levels in blood cells measured by enzymatic assay

Direct glucosone-based synthesis and HILIC-ESI-MS/MS characterization of N-terminal fructosylated valine and valylhistidine for validation of enzymatic HbA1c assays in the diagnosis of diabetes mellitus

Pengaruh Lama Penyimpanan 5 Jam dan 10 Jam pada Suhu 2-8 0C Terhadap Kadar Glycated Hemoglobin (HbA1c)

Contact Info

Product

Resources

About