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Blood-, urine- and liquor sugar II
BRIEF
NOTES
Blood-, urine- and liquor sugar II In 1963 we published a rapid determination for sugar in blood, urine and liquor, employing 3,5_dinitrosalicylic acid (DNS)l. A follow up study has been made comparing this method with the N-method of AutoAnalyser for glucose. Because of small modifications the reagents and methods are mentioned briefly. .METHODS
AutoAnalyser (Technicon), blood and I ml NaCl 0.9%).
N-method
for glucose (K,Fe(CK),,
sampling
0.1 ml
Reagents fey the DNS assay (A) 1.5”/ DNS (B.D.H.) in H,O. (B) Add 13.8 g phenol and 13.8 g NaHSO, tartrate in 880 ml H,O. (C) 400 ml A+80 ml H,O+480 ml B.
in 350 ml NaOH
10% to 510 g Na-K-
Procedure for blood and liqzcor Blow 0.1 ml blood or liquor into I ml of reagent .4, mix on a Vortex mixer, centrifuge for I min 2000 rev./mm, pipet 0.5 ml of clear supernatant and add 0.5 ml of reagent B, place 3 min in a boiling water bath and add 4 ml H,O (autodilutor). Procedure fey wine 0.01 ml urine is mixed with I ml reagent C (ultramicro place 3 min in a boiling water bath and add 9 ml H,O. Readings at 540 nm against H,O as a blank. RESULTS
AND
pipet or autodilutor),
COMMENTS
Besides regular routine checks with patient blood sugars a comparison was made during 5 years with 47 unknown sera (courtesy of Dade Reagents, Miami, Fla.) in the range of 4.1-17.8 mmole/l. The following mean values were found: Dade Reagents AutoAnalyser 7.5 mmole/l Folin-Wu 7.9 >> Present
work
AutoAnalyser DNS
7.5
,,
7.3
!J
Shats et aZ.2 published almost this same method, employing 0.1 ml of blood and omitting YaHSO, in the reagent B. They state erroneously that our DNS method has previously been used for urine. We feel that NaHSO, is a necessity for stability of the color. The DNS solution coagulates the blood proteins strongly, the pipets should therefore not be rinsed with this solution but put into a detergent after blood sampling. A high protein level in liquor can already be noticed when doing the sugar test. The method has proven very useful in general, also for glucose tolerance tests, and can be used with good success in small hospitals which cannot afford automation. czin. Chsm. Acta, 34 (197’)
489-.+90
490 De$artment of Clikcal
Chemistqf,
St. Elisahrth Hosfiital Willemstad, Curapao (Netherlands Antilles)
Received
March 3, 1971
czm.Chit%. Acta, 34 (1971) 439-490
A. I..GITERSON I..A. s. 1,IE KONG I. E. WEJ)ENF~OIU H. SCHOUTi-K
NOTES
Blood-, urine- and liquor sugar II In 1963 we published a rapid determination for sugar in blood, urine and liquor, employing 3,5_dinitrosalicylic acid (DNS)l. A follow up study has been made comparing this method with the N-method of AutoAnalyser for glucose. Because of small modifications the reagents and methods are mentioned briefly. .METHODS
AutoAnalyser (Technicon), blood and I ml NaCl 0.9%).
N-method
for glucose (K,Fe(CK),,
sampling
0.1 ml
Reagents fey the DNS assay (A) 1.5”/ DNS (B.D.H.) in H,O. (B) Add 13.8 g phenol and 13.8 g NaHSO, tartrate in 880 ml H,O. (C) 400 ml A+80 ml H,O+480 ml B.
in 350 ml NaOH
10% to 510 g Na-K-
Procedure for blood and liqzcor Blow 0.1 ml blood or liquor into I ml of reagent .4, mix on a Vortex mixer, centrifuge for I min 2000 rev./mm, pipet 0.5 ml of clear supernatant and add 0.5 ml of reagent B, place 3 min in a boiling water bath and add 4 ml H,O (autodilutor). Procedure fey wine 0.01 ml urine is mixed with I ml reagent C (ultramicro place 3 min in a boiling water bath and add 9 ml H,O. Readings at 540 nm against H,O as a blank. RESULTS
AND
pipet or autodilutor),
COMMENTS
Besides regular routine checks with patient blood sugars a comparison was made during 5 years with 47 unknown sera (courtesy of Dade Reagents, Miami, Fla.) in the range of 4.1-17.8 mmole/l. The following mean values were found: Dade Reagents AutoAnalyser 7.5 mmole/l Folin-Wu 7.9 >> Present
work
AutoAnalyser DNS
7.5
,,
7.3
!J
Shats et aZ.2 published almost this same method, employing 0.1 ml of blood and omitting YaHSO, in the reagent B. They state erroneously that our DNS method has previously been used for urine. We feel that NaHSO, is a necessity for stability of the color. The DNS solution coagulates the blood proteins strongly, the pipets should therefore not be rinsed with this solution but put into a detergent after blood sampling. A high protein level in liquor can already be noticed when doing the sugar test. The method has proven very useful in general, also for glucose tolerance tests, and can be used with good success in small hospitals which cannot afford automation. czin. Chsm. Acta, 34 (197’)
489-.+90
490 De$artment of Clikcal
Chemistqf,
St. Elisahrth Hosfiital Willemstad, Curapao (Netherlands Antilles)
Received
March 3, 1971
czm.Chit%. Acta, 34 (1971) 439-490
A. I..GITERSON I..A. s. 1,IE KONG I. E. WEJ)ENF~OIU H. SCHOUTi-K