FINALS NOTES 1

HUMAN PLACENTAL LACTOGEN – pregnancy-related hyoerglycemia days)

HUMAN CHORIONIC SOMAMOTROPIN – similar to action of GH FORMATION OF HB A1C
(inhibits insulin)
1 Glucose + Valine -------- [N-terminal of B]
GESTATIONAL DIABETES MELLITUS 2 SCHIFF BASE (unstable aldimine) -------- [Amadori
 Glucose intolerance which develops in 7% of pregnancies rearrangement]
 Abnormal glucose concentration discovered for the 1st time 3 KETOAMINE (stable)
during pregnancy
NOTES:
GDM SCREENING & DIAGNOSIS (ADA 2017)
 GHb testing provides an index of ave. BGL over the past 2-4
months
ONE-STEP STRATEGY (2011 Standards of Care – ADA)
 Values for total HbA1 & HbA1c have a high degree of
 24 to 28 weeks of gestation
correlation
 Step 1: Perform a 75-g OGTT (fasting, 1H, and 2H BGL)
 Labile fraction (Schiff base or pre-A1c): accounts for 5 to 8% of
Diagnosis of GDM is made when any or the PG values are = or > total HbA1
than any of the ff PG levels:  Advantages:
Fasting: 92 mg/dL (5.1 mmol/L) 1. Provides objective means of reflecting control of DM
1 h: 180 mg/dL (10.0 mmol/L) 2. Additional confirmation of the clinical impression of
2 h: 153 mg/dL (8.5 mmol/L) control

TWO-STEP STRATEGY (2013 – NIH) METHODS FOR DETERMINATION

 24 to 28 weeks of gestation
 Step 1: Perform a 50-g GLT (non-fasting)
1. ION-EXCHANGE 2. HPLC
o 1H PG measurements
CHROMATOGRAPHY
 If the 1H PG is > or = to 130 mg/dL, or
 Cation-exchange resin or  Reference method
140 mg/dL, proceed to a 100-g OGTT carboxymethyl cellulose  Separates and quantifies
 Step 2: The 100-g OGTT should be performed when the resin HbA1c and other
patient is fasting  HbA1 elutes from the haemoglobin types
Diagnosis of GDM is made if at least two values are = to or > than column first
any of the ff PG levels:  False elevation: labile
Fasting: 105 mg/dL fractions, HbF
1 h: 190 mg/dL  Low values: Hb variants
2 h: 165 mg/dL 3. COLORIMETRY 4. RADIOIMMUNOASSAY (RIA)
3 h: 145 mg/dL  Hb A1c --- acid --- 5-HMF  Antibodies against Hb A1c
 Specific for ketoamine- (sheep antiserum)
CLINICAL SIGNIFICANCE linked  Partial cross-reactivity w/
 Unaffected by HbF, Hb HbA1c
HYPOGLYCEMIA variants and labile  Antiserum not
 Results from an imbalance between glucose utilization and intermediate commercially available
production  Routine
 Observable symptoms appear at about 50 to 55 mg/dL 5. ELECTROPHORESIS 6. ISOELECTRIC FOCUSING
Citrate agar electrophoresis  Scanned on high-
SYMPTOMS (pH 6.0-6.2) resolution
- Buffer: Citrate agar buffer microdensitometer
(acidic)  Hb A1c is adequately
NEUROGENIC/ADRENERGIC NEUROGLYCOPENIC - Solid support: agarose gel resolved from HbA1a,
(EPINEPHRINE) *HbF migrates to the A1b, S and F
 Triggered by the ANS  CNS hypoglycemia same region as HbA1 - Medium: Polyacrylamide
 Tremulousness,  Dizziness, tingling, Cellulose acetate gel
palpitations, anxiety difficulty in concentrating, electrophoresis (pH 8.0)
 Diaphoresis (cold sweat) blurred vision - Buffer: Tris EDTA boric
 Hunger  Confusion, behavioral acid (alkaline)
 Paresthesias (numbness) changes, seizure, coma
- Solid support: cellulose
acetate
TESTS FOR MONITORING DIABETIC PATIENTS
 Good resolution of Hb A &
Hb A1
MEASUREMENT OF HbA1  No interference from Hb
 Glycohemoglobin aka fast Hgb, glycosylated Hgb variants
 HbA1c – major fraction; predominant form  HbF migrates to the same
o Non-enzymatic condensation between glucose and region as HbA1
the N-terminal valine of each B-chain
o Occurs over the lifespan of the RBC (90 to 120

1
FINALS NOTES 1

7. AFFINITY CHROMATOGRAPHY
 Use of affinity gel columns
 Advantages:
o No interference from non-glycosylated Hb
o Negligible interference from the labile
intermediate
o Minimal dependence on variations in ambient T

REFERENCE RANGES

MEAN % RANGE %
Total Hb A1 6.5 5.0 – 8.0
Hb A1c only 4.5 3.0 – 6.0

CORRELATION BETWEEN GHB AND PLASMA GLUCOSE LEVEL

HbA1c (%) Approximate Plasma Glucose

mg/dL Mmol/L
5 97 5.4
6 126 7.0
7 154 8.6
8 183 10.2
9 212 11.8
10 240 13.4
11 269 14.9
12 298 16.5

MEASUREMENT OF GLYCOSYLATED PROTEIN OR GLYCOSYLATED

ALBUMIN

 For patients with inaccurate HbA1c assays (hemglobinopathies

and hemolytic anemias)
o Affinity chromatography
o Immunoassays
 Turnover time of serum proteins: (primary albumin) 14 to 20
days
o Reflects glycemic control over narrower period of
time

FRUCTOSAMINE ASSAYS
 Most widely used to assess short-term (3 to 6 week) glycemic
control
 Advantage: Use of serum samples and automated equipment
(simple to perform and low in cost)
 More reliable than other glycosylated protein assays