UNIVERSITY of SAN AGUSTIN

WILHELM C.
LIZADA, RMT
Professional Service and Applications Representative
CAPILLARY ELECTROPHORESIS TECHNOLOGY:

A Breakthrough in HBA1C, Hemoglobin, Thalassemia and Serum
Protein for the Determnation of Pathologic Conditions
UNIVERSITY OF SAN AGUSTIN

Iloilo City, 5000
September 27, 2023 1:00-3:00 PM
TOPICS
• Capillary Electrophoresis Test and Application

• Clinical / Laboratory Cases
• How Hemoglobin Disorder (Hemoglobinopathies
and Thalassemia) affect HbA1c Result
• Diabetes and Methodology and Standardization
• Global Impact and Article Journal Studies and
Statistical Studies
• Take Home Message
• Installed Base (2023)
INTRODUCTION ON
ELECTROPHORETIC TECHNOLOGIES
CAPILLARYS
3
OCTA
CAPILLARYS 3 OCTA
A large specter of diseases analyzed by
Capillary Electrophoresis technologies
Capillarys Electrophoresis Technology
What is CAPILLARY ELECTROPHORESIS?
It is a TECHNOLOGY… It is a Top of the LINE TECHNOLOGY…
MASS SPECTROMETRY TECHNOLOGY
PCR (POLYMERASE CHAIN REACTION)
STRICTLY CONFIDENTIALTO SEBIA - NOT

FOR RELEASE
CAPILLARY ELECTROPHORESIS TECHNOLOGY
• Capillary electrophoresis is an analytical technique that separates

ions based on their electrophoretic mobility with the use of an
applied voltage. The electrophoretic mobility is dependent upon
the charge of the molecule, the viscosity, and the atom's radius.
The rate at which the particle moves is directly proportional to the
applied electric field--the greater the field strength, the faster the
mobility. Neutral species are not affected, only ions move with the
electric field. If two ions are the same size, the one with greater
charge will move the fastest. For ions of the same charge, the
smaller particle has less friction and overall faster migration rate.
Capillary electrophoresis is used most predominately because it
gives faster results and provides high resolution separation. It is a
useful technique because there is a large range of detection
methods available.
FOR RELEASE
MS - Identification of unknown compounds:
Mass spectrometry helps in identifying the Clinical diagnostics: CE plays a role in
components of a mixture by analyzing the clinical applications such as analyzing
mass-to-charge ratio (m/z) of ions produced genetic disorders, identifying disease
from the sample. It can identify organic biomarkers, and determining drug levels
compounds, biomolecules, drugs, and in patient samples.
pollutants in complex mixtures.
FOR RELEASE
Principle of HbA1c (also HbE & SPE)
using capillary electrophoresis technologies
Injection of
Detection proteins
+ + + + + + + + + + + + + + + + + +
---
--
- ---
--
+
Cathode -
--- Anode
--
-
+ + + + + + + + + + + + + + + + + +
Electro-endosmotic flow Electrical field
The Electro-Osmotic Flow (EOF) is a stronger force than the Electrical

Field. As a result, all proteins are carried towards the cathodic end of
the capillary
Main Applications of Capillary
Electrophoresis
Serum & Urine Proteins
Screening and monitoring of
Multiple Myeloma &
Pathological Condition
Hemoglobin
Screening of thalassemia &
hemoglobinopathies
HbA1c Screening and

monitoring of Diabetes
TEST AVAILABLE in
CAPILLARY ELECTROPHORESIS
1. Serum Protein Electrophoresis (SPE)

2. Urine Protein Electrophoresis (UPE)
3. Immunotyping Test (IT)
4. Carbohydrate Deficient Transferrin (CDT) – Chronic
Alcohol Abuse
5. Hemoglobin Electrophoresis (Hgb E)
6. Hb A1c Electrophoresis (Hb A1c Elec. Test)
Request and Test
TEST REQUESTED
HbA1c / SPE
UPE / SPE / IT
HbA1c
UPE / SPE
HBE
HBE
SPE
SPE
HBE / HBA1C
SEBIA
(from Paris, France)
50 years at the service of MM diagnosis
Multiple Myeloma / Monoclonal Gamopathies
thru
Serum Protien Electrophoresis Test
FOR RELEASE
Serum Protein Electrophoresis (SPE)
for Monoclonal Gammopathies (Plasmocyte Cancer) & Pathological
Condition Screening & Confirmation (Chronic and Acute Inflammation)
(Multiple Myeloma,MGUS,SM, Lymphoma etc. Liver Cirrhosis, Nephrotic Syndrome, Renal
Failure)

FOR RELEASE
Serum protein electrophoresis and detection of monoclonal
immunoglobulin
P1 P2 P3 P4 P5
Normal
plasmocytes
Polyclonal immunoglobulins
Malignant transformation
of a plasmocyte
Polyclonal
Immunoglobulins Qualitative and/or
Monoclonal quantitative modifications of
immunoglobulin serum protein electrophoresis
Multiple Pathologies TP, Split β-
screening g/dL
Alb α-1 α-2 β β bridge 
Acute inflammatory pattern and

Hyper-estrogenism pattern   
Chronic inflammatory disorders     
Hypergammaglobulinemia 
Liver disease/Cirrhosis   + 
Nephrotic syndrome    
Protein-losing pattern/malnutrition      
Autoimmune reactions   
Alpha-1-antitrypsin deficiency 
Hypogammaglobulinemia  
Hemolytic anemias/congenital
hemoglobin defects 
Total protein, Albumin, alpha 1 & 2 zones, beta zone, gamma zone (gammaglobulins)
The most complete offer on the market with no equivalent elsewhere
IT
UIT
IF plus *
Serum
Free light
SPE
chains
UPE
IF
SebiaFLC
BJ Seralite
HYDRASHIFT
daratumumab
Screening Peak/BJ typing Precision medicine

High sensitive IF sFLC quantification
Peak quantification Complete response typing (monoclonal antibody IV)
Peak monitoring Kidney lesion typing
SARCLISA®
(Isatuximab)
NINLARO®
( Ixazomib )
Urine Protein Electrophoresis (UPE) – for complimentary test of SPE, for
confirmation & screening as well. Screen and Typing for Bence Jones
Protein or Monoclonality. Renal Failure Typing for possible kidney
damage and identify if (tubular, glomerular, overload or mixed)

FOR RELEASE
Unconcentrated urine Minicap Urine
Urine electrophoresis
SEBIA
dialysis Identification of a monoclonal peak
system
Concentrated urine
Minicap Urine IT
identification of a BJ
Analysis within 24 hrs on Minicap

Immunotyping Test (IT)
- Detection of monoclonal protein on electrophoresis with specific antisera

- Confirmation of monoclonality
- Typing of monoclonal proteins visible on SPE
If SPE is normal but there is a strong clinical suspicion of myeloma,
immunotyping or immunofixation should be preferred

FOR RELEASE
10 different immunoglobulin combinations
IgA Kappa IgD Kappa
IgM Kappa IgE Kappa

IgG Kappa
IgA Lambda
IgD Lambda
IgG Lambda IgM Lambda IgE Lambda

Immunotyping - Immunosubtraction
Abnormal peak The peak disappears in Ig G

in gamma
Immune complex
The peak disappears in Kappa
Conclusion: Detection of monoclonal Ig G Kappa

Strictly confidential to Sebia
Carbohydrate Deficient Transferrin (CDT) –
Chronic Alcohol Abuse. The Machine could
detect Alcohol on the past 30days.
Hemoglobin Electrophoresis (Hgb E)
– Hemoglobinopathies (Sickle Cell, Hgb Variants) & Thalassemia's (Alpha or Beta).

- FOR NEWBORN SCREENING
- FOR ADULT

FOR RELEASE
GLOBAL IMPACT
HEMOGLOBIN CAPILLARY
ELECTROPHORESIS
Thalassemia
a global health issue
Hemoglobinopathies,
a global health problem...
Hemoglobinopathies (Hb variants)
Births with a pathological hemoglobin disorder (per 1,000 live births)

Adapted from WHO, 2008
Thalassemias
Immigration Trends Impacting Thalassemia

Adapted from http://www.ironhealthalliance.com
NEWBORN SCREENING PREVALENCE GLOBALLY –
International Journal Laboratory Hematology 2016
27% - SEA
Alpha Thalassemia with Hgb H

(Filipino – 15%)
Distribution of Hb disorders Worldwide
Hemoglobinopathies have been recognized
as a public health priority by the World Health Organization
WHO Resolution on Sickle Cell

Anemia
WHO Resolution on thalassemiaand

other hemoglobinopathies
UNESCO Resolution on Sickle Cell

Anemia
UN Recognition of Sickle Cell as a pu

blic health problem
Newborn Screening Center & Confirmatory for
Hemoglobinopathies & Thalassemia, Manila, Philippines
• Data and Cases (Jan. 2015 – Aug. 2017) as per NIH Newborn
Screening and Confirmatory): not yet subsidized by Philhealth
• 156,000 test on screening
• 2,110 test (positive for HbP & Thal)
• Ratio is 1:74
• (4,791 baby born per day / 200 baby per hour) = PSA 2014
• = 65 baby positive / day = 1,950 / month = 23,400 / year
70,000 – 95,000 per year
• #1 – Hb E - (15%)
• #2 – HbH (alpha thal) – (11%)
• #3 – Alpha Thal Trait – (10%)
• #4 – HbD – (6%)
• #5 – HbC – (4%)
• #6 – HbE Beta Thal – (2%)
• #7 – HbS (Sickle Cell Trait) – (0.3%)
(data with the permission of NIH Newborn Screening & Confirmatory Center – 2017 @ per Dr. De Castro N.- Hematologist) /
Exact Data for the Positive Case is Confidential under NIH)
RANKING BASED ON
PREVALENCE
(NEWBORN SCREENING PHILIPPINES)
 1. G6PD DEFICIENCY 3 – 5 years as per

NIH, NSC
 2.HEMOGLOBINOPATHIES AND Predicted
THALASSEMIA
 3. CONGENITAL HYPOTHYROID
 4.CONGENITAL ADRENAL
HYPERPLASIA
 5.FATTY ACID DISORDERS
INTERESTING LOCAL
CASES (Hemoglobin Cases)
CASE & DATA
• PATIENT IS PREGNANT
• TEST FOR HbA1c – RESULTS is low
HbA1c = 3.8%
• Baby Died during Pregnancy due to
“Hydrops Foetallis” (both parents are
alpha thal)
• Clearly she is Alpha Thallasemia – HbA2 =
0.7%, Hb (1) “Hgb H” = 6.0%
• Husband is a suspected Alpha
Thalassemia carrier
Hgb Electrophoresis Test -
Confirmatory
Hematological Parameters
- Hb 11.5 g/L (115 – 145)

- MCV 64 fL (80 – 100)
- MCH 18.80 pg (24 – 30)
Anemia, Microcytosis, Hypochromia
Hgb Electrophoresis Fraction
Hb H – 5%
Hb A – 94.25 %
Hb A2 – 0.75%
The presence of Hb H, reduced HbA2,

anemia, and microcytosis indicate an
alpha-thalassemia
Hgb Electrophoresis Test

Iron / Ferritin Result
Ferritin Result – 139.7ng / Ml

Range (13 – 150) ng/mL
Ferritin Level
Peripheral Blood Smear
Teardrop Cells,
Target Cells
INTERPRETATION /
CONCLUSION: ALPHA
THALASSEMIA WITH Hgb H
Hgb Barts Hydrops Fetalis
• IN CASE WE GET THE SAMPLE OF THE
BABY…WE USUALLY SEE THIS
CURVE.. Hgb Barts
Gower 1
Epsilon 4
Hgb
Portland
Why We Need Hemoglobin
Electrophoresis Test?
1 OUT OF 4 CHILD
25%
Alpha - Thalassemia
Beta - Thalassemia
Hemoglobin Electrophoresis
DETECTION OF DIFFERENT HGB VARIANT
STRAIGHTFORWARD DETECTION OF
THALASSEMIA
Result display
Automatic presumptive
identification of the fractions
Result display in Phoresis 9.15
Migration zones of 525 Hb variants in the database

(vs. 348 variants in Phoresis 8.63 )
Library of Hb variants in Phoresis
Z 15: Hb H, Hb I-Texas
software
Z 14 : Hb N-Seattle
Z 13 : Hb-J Rovigo, Hb N-Baltimore…
Z 12 : Hb Bart’s, Hb J-Providence, Hb J-Mexico, Hb J-Baltimore….
Z 11 : Denatured Hb A, Hb Kaoshiung…
Z 10 : Hb Hope, Hb M-Iwate…
Z(A) : Hb A, Hb Camperdown, Hb Phnom Penh……
Z 8 : Acetylated Hb F, Hb Altanta, Hb Athens-GA…
Z (F): Hb F, denatured Hb S, Hb Porto-Alegre……..
Z(D): Hb D-Punjab, denatured Hb E, Hb Korle-Bu, Hb Lepore, Hb Köln….
Z(S): Hb S, Hb Hasharon, Hb Handsworth, denatured Hb O-Arab…
Z(E): Hb E, denatured Hb C, Hb Köln, Hb A2 variants, M-Iwate Hb A2
variants…
Z(A2): Hb A2, Hb O-Arab…
Z(C): Hb C, Hb Constant Spring, Setif HbA2 variant…
Z 1: Hb dA2 Hb aA2, Hasharon Hb A2 variant, Winnipeg Hb A2 variant……
Hemoglobinopathies and Diabetes
• Worldwide population: 7,5 billions approx.
HbA1c Test
Diabetes
Prevalence Type II = 5.8%
463 millions of people affected
We always refer to the diabetes epidemy...
Data from WHO and

IFD - 2019
Hemoglobinopathies and Diabetes
• Worldwide population: 7,5 billions approx.

Hgb E, Alpha-Thal, Beta Thal,
Sickle Cell Hemoglobinopathies
Prevalence = 7.2%
HbA1c Test
Diabetes
Prevalence Type II = 5.8%
... while we are still considering

hemoglobinopathies as an orphan disease!
Data from WHO and
IFD - 2019
How to diagnose Diabetes?
Criteria for the diagnosis of Diabetes
Fasting plasma glucose FPG ≥ 126 mg/dL

(FPG) (7.0 mmol/L)
Or
Oral Glucose Tolerance Test 2-h PG ≥ 200 mg/dL
(OGTT) (11.1mmol/L) during an OGTT
Or
Patient with classic Random plasma glucose ≥
hyperglycemia symptoms or 200mg/dL (11.1 mmol/L)
Or hyperglycemic crisis
Hemoglobin A1c Hb A1c ≥ 6.5%
(Hb A1c) (48 mmol/mol)
64
What is Hb A1c?
 Hb A1c: biomarker of Diabetes
 Hb A1c: measure of hemoglobin A glycation
 Hb A1c reflects mean glycemia for the previous 3 months

(red blood cell lifespan is about 120 days)
 Hb A1c provides clinicians information about a long-term glycemic

control
65
Hemoglobins in blood
α1 β1 α γ1 α1 δ1
1
β2 α2 γ2 α2 α2
δ2
Hb A > 96.5% Hb F ≤ 0.5% Hb A2 < 3.2%
67
Non-Enzymatic Glycation of Hb A
Stable
Labile HbA1c
Definition of the Analyte
according to IFCC:
α1 β1
How is Hb A HbA1c is biochemically
glycated with glucose? characterised as the
stable adduct of glucose
to the N-terminal amino
group of the β-chain of
β2 α2 hemoglobin A0
12stndStep:
Step:Unstable,
During redreversible
blood cellreaction between
circulation, someGlucose and the
of the labile A1cN-
is
terminal
convertedvaline of the
to form β-chain
a stable (Schiff
HbA1c base) rearrangement)
(Amadori
68
The nature of the problem – what is HbA1c?
HbA1c is currently defined as:

Hemoglobin A which is irreversibly glycated at one or both N-terminal
Valines of the  chains in the tetramer.
Glycation elsewhere on the  or  chains is irrelevant.
G G
   
 
    N G  
N N
G G G
G  
N
  G
N
G
All of these are HbA1c

The nature of the problem – what is HbA1c?
Glycohemoglobin, or GHb, or Total GHb, is defined as:

Hb having one or more sugars irreversibly attached at any point in any
of the globin chains.
(This also includes all forms of HbA1c).
G
  G
G G   N
G  
    N
G
 
N   G
G   G
N
All of these are GHb (but not HbA1c)

Hb A1c: What are we looking for?
Hb A Hb A0
93-95%
Glycation at the
N-terminal Valine
of the β-globin chain
Hb A1 = GHb
Glycated HbA
5-7% Hb A1a Hb A1b Hb A1c
0.5% 0.5% 4-6%
Fructose-1,6- pyruvate glucose
diphosphate
Hb A2Glucose-6-phosphate
Hb F
+ + + 71
HbA0
HbA1c
HbA1a+1b+3 HbA2
HbF
HbA1c – C.E.
Diabetes and Hb A1c level
Hb A1c values from by American Diabetes
Association (ADA)
Elevated Hb A1c =
diabetic patient
73
Diabetes diagnosis and monitoring
 Hb A1c: initially used only for the monitoring of Diabetes
 New guidelines that promoted Hb A1c as a diagnostic test:
American Diabetes Association guidelines 2010
World Health Organization guidelines 2011
74
Diabetes Testing – HbA1c Testing
Glycosylated Hemoglobin
HbA1c – C.E.

HOW ACCURATE and PRECISE
THE RESULT IS?
WHICH METHOD SHOULD I USED?
POINT OF CARE - POC
HPLC
IMMUNASSAY/
SEBIA Capillary Electrophoresis Technologies
Your new solution
for
the analysis
of
Hb A1c Electrophoresis
(The Latest INNOVATIVE Technology for HbA1c Testing)

Clinical Cases:
Capillary
Electrophoresis Vs.
Other Method
(Technology use in
the Laboratory)
Why is it Preferred by the Clinicians,
Endocrinologist, Diabetologist and
Pathologist
To request / HbA1c Test using
Capillary Electrophoresis
Technology?
HbA1c Electrophoresis
“ Breakthrough in Hb A1c Testing ”
(Capillary Electrophoresis Technology)
- Precise and Accurate Result without any Analytical & Biological

Interference
- Screening for Hemoglobinopathies & Thalassemia
CLINICAL CASES:
INTERESTING CASEs
(International)
Case description
66yrs old spanish male patient with type II diabetes history
Patient Highest reference Conclusion
limit value
Glycemia 8,1 mmol/l 6,1 mmol/l Positive
Fructosamine 290 µmol/l 285 µmol/l Positive
Système HbA1c Variant detection Conclusion
BioRad Variant II Turbo 5.3% Yes Negative
Arkray HA-8160 4.3% No Negative

Roche Tina-Quant 6.2 % No Negative
BioRad In2it 5;9% No Negative
Sebia Capillarys 2 FP 6.8% Yes Positive
HPLC UNDERESTIMATE HbA1c
Result due to Hgb Variant “Hgb
Jerez” by HPLC Formula
HPLC
Fasting glucose : 8.1mmol/l (upper limit

reference interval 6.1mmol/l)
Fructosamine: 290 µmol/l (upper limit
reference interval 285 µmol/l )
C. E.
Analytical interference on HPLC
%HbA1c =
[100 x (HbA1c area / ∑ HbA area)]
∑ HbA area = ∑ ( HbA1a11/a22 + HbA1b + LA1c

HbA1c + P3 + HbA0 )
+ HbA1c
LA1c & HbA0 are part of the ∑ HbA area
in which the Hgb Jerez co-elutes
(This underestimate or lowered the HbA1c Value)

CLINICAL CASES:
INTERESTING CASEs
(Phils. / Local)
HbA1c – Underestimation of the Result
C.E. vs. JAPANESE HPLC
HbA2 –
Indicator for
Thalassemia
HbA1c – 6.5% Diabetic Cut Off Value

Hgb Variant Straight Forward Detected
CBC Result
Anemia – Hgb = 7.0 g/dL
Microcytosis, Hypochromia
HPLC RESULT
Area – less than 800
No HbA1 Result – (Dashes)

Clinical context (Local Case)
15-yrs old female, from Western Visayas

With a physical description of pale skin.
Mother is a Beta Thalassemia Trait
Father is a Hgb E Heterozygous Carrier
Having blood transfusion on a monthly basis with
Iron Chelation therapy afterwards.
C.E. (Capillary Electrophoresis) Vs. Chinese HPLC
MICROCYTIC
ANEMIA
Clinical context (Local Case)
15-yrs old female, from Western Visayas
With a physical description of pale skin.
Mother is a Beta Thalassemia Trait
Father is a Hgb E Heterozygous Carrier
Having blood transfusion on a monthly basis with Iron
Chelation therapy afterwards.
• Hgb A – 70.7 (Normal Range 96.5

above)
• Hgb E – 18.6 %
• Hgb F – 7.2 %
• Hgb A2 – 3.5 %
• Hb 8.9 g/L (115 – 145)

• MCV 76.9 fL (80 – 100)
• MCH 24.3 pg (25 – 30)
Chronic Anemia, Microcytosis,

Hypochromia
Hgb E Heterozygous / Beta Thalassemia

Confirm with Hemoglobin Electrophoresis Testing
• Hgb A – 70.7 (Normal Range

96.5 above)
• Hgb E – 18.6 %
• Hgb F – 7.2 %
• Hgb A2 – 3.5 %
• Hb 8.9 g/L (115 –

145)
• MCV 76.9 fL (80 – 100)
• MCH 24.3 pg (25 – 30)
Chronic Anemia, Microcytosis,

Hypochromia
C.E. (Capillary Electrophoresis) Vs. Chinese HPLC
Hgb Variant Detected
7.0 % ?
?
Elevated HbA1c - Diabetic
6.1 %
HPLC
HbA1c – C.E. (Capillary Electrophoresis)
HPLC UNDERESTIMATE HBA1C VALUE

in the Presence of Hgb Varint
CLINICAL CASES:
INTERESTING CASEs
(Phils. / Local)
REFERENCE CENTER / GOVERNMENT
UNIVERSITY HOSPITAL – MANILA
(capillary electrophoresis vs. hplc)
CAPILLARY
ELECTROPHORESIS
Hgb Variant HPLC
co – elute with
5.8 %
6.5 %
Analytical interference on HPLC
%HbA1c =
[100 x (HbA1c area / ∑ HbA area)]
∑ HbA area = ∑ ( HbA1a11/a22 + HbA1b + LA1c

HbA1c + P3 + HbA0 )
+ HbA1c
Hgb E Variant co-elutes HbA1c area
(This overestimate or falsely elevated result of the

HbA1c Value)
TAKE NOTE FOR THE
RESULT
• CAP2FLEX = 5.8% (NORMAL PATIENT)
– HBE E VARIANT DETECTED
– IFCC / NGSP REFERENCE RANGE
– NORMAL RANGE (4.0 – 6.0) % & (20 – 42) mmol / mol
– DIABETIC (6.5 onward) %
• HPLC = 6.5% (HIGH RESULT / DIABETIC) ???

• CHECK WITH CHEMISTRY FBS = 5.5mmol/L
• - NORMAL RANGE (FBS) 5.6 to 6.9 mmol/L
• “PATIENT HBA1C IS NORMAL”

Private Hospital – MANILA
(CAPILLARY ELECTROPHORESIS VS. CHEMISTRY MACHINE /
IMMUNOASSAY)
FOETAL HGB
Black box – no idea for

hemoglobin concern 5.2%
6.6% Falsely Elevated Result

Diabetic – note: 6.5% cut off value
DAY 3: JUNE 29, 2015
SAMPLE
NO
PATIENT ID C.E. results
CAPILLARYS 2FLEX RESULT vs.D10 (HPLC)
hplcRESULT
mmol/mol cal % cal %
1 150071906110 43 mmol/mol 6.1% 6.1%
2 150071895110 51 mmol/mol 6.8% 6.9%
3 150071924110 43 mmol/mol 6.1% 6.1%
4 150071899110 54 mmol/mol 7.1% 7.2%
5 150071943110 106 mmol/mol 11.9% 11.5%
6 150071907110 48 mmol/mol 6.6% 6.7%
7 150071817110 47 mmol/mol 6.5% 6.5%
8 150071945110 38 mmol/mol 5.6% 5.8%
9 150071900110 86 mmol/mol 10.0% 9.9%
10 150071892110 58 mmol/mol 7.5% 7.4%
11 150071902110 44 mmol/mol 6.2% 6.2%
12 150071905110 37 mmol/mol 5.6% 5.8%
13 150072023110 45 mmol/mol 6.3% 6.5%
14 150072048110 37 mmol/mol 5.5% 5.8%
15 150071822110 36 mmol/mol 5.5% 6.1%
16 150071809110 39 mmol/mol 5.8% 6.0%
17 150071803110 50 mmol/mol 6.7% 6.7%
18 150072104130 115 mmol/mol 12.7% 12.7%
SIEMENS
C.E. results vs. immunoassay
SAMPLE
PATIENT ID CAPILLARYS 2FLEX RESULT (DIMENSION EXL)
NO
RESULT
1 150071735110 106 mmol/mol 11.9% 15.2%
2 150071825110 30 mmol/mol 4.9% 6.9%
3 150071873110 33 mmol/mol 5.2% 6.6%
4 150071865110 40 mmol/mol 5.8% 7.1%
5 150071859110 31 mmol/mol 5.0% 7.1%
6 150071855110 50 mmol/mol 6.7% 8.3%
7 150071831110 46 mmol/mol 6.3% 7.9%
8 150071874110 36 mmol/mol 5.4% 7.1%
9 150071828110 41 mmol/mol 5.9% 7.6%
10 150071843110 38 mmol/mol 5.7% 6.6%
11 150071866110 39 mmol/mol 5.7% 7.2%
ORANGE – DIABETIC, WHITE –
NORMAL, PUPLE WITH Significantly high
VARIANT, YELLOW
(ELEVATED-SIGNIFICANT)
Private Laboratory in BINONDO
CAP2FLEX VS.HPLC
Hgb
Kaoshiung
Interesting Case
• In Summary
– COMPARISON OF HPLC AND CAP2FLEX

(SEBIA) – MACHINE
– SAMPLE WITH HBG VARIANT Hb
KHAOSHIUNG
– RESULTS:
– CAP2FLEX = 5.7%
– HPLC = 6.4%
– NOTE: HPLC MACHINE IS HIGHER CANNOT SEPARATE THE Hb
KHAOSHIUNG
NORTHERN
MINDANAO
Hgb
Variant 1st Run:
HbA1c – C.E. 12.4 % HPLC
Falsely Elevated Result !!
4.9 %
2nd Run:
20.4 %
Hgb Electrophoresis:
Hgb Variant - Hgb E Heterozygous

Hgb E = 14.7%
CBC RESULTS:
MCV = 70 (79.7 – 97.0) U

MCHC = 30.6 (32.2 – 35.0) g/ Dl
GLUCOSE RESULTS:
FPG/FBS = 90 mg/dL (< 99 mg/dL)

OGTT = 127 mg/Dl (< 139 mg/Dl)
HPLC
HbA1c – C.E.
Hgb Variant & Thalassemia Detection – No
Interference with HbA1c Result
6.0 %
HbA1c is False Elevated due to
Hgb Variant co-elute with
HbA1c. Cannot Detect Hgb
Variant
7.8 %
Private Hospital in Zamboanga
- Pathological Case -
A TYPICAL HEMOGLOBIN E HOMOZYGOUS (HbE HOMOZYGOUS) – HgB
Electrophoresis is recommended
No HbA1c
Value
Hemoglobin molecule is a tetramer
Glucose attached to the N-Terminal Valine of the Beta Globin
4 polypeptide chains G
(globins)
of 2 different types β (beta), δ (delta) or
γ (gamma)
α (alpha)
O
2
binding and transport

heme group bound to
of oxygen
each globin chain
Sebia, Hb interpretation I 111

Formation of hemoglobin molecules
Hb A
Hb A
α2β2 β β
Hb A2
α α
 
Hb A2 Hb A
α α α2δ2
 
Hb   Hb F Hb A2
F
α2γ
2

Hemoglobin by Capillary
HbA1c by Capillary Electrophoresis
Electrophoresis
Glucose attached to the N-

HbA0 HbA Terminal Valine
(α2β2) of the Beta Globin
α1 β1
β2 α2
G
HbA1c HbA1a+1b+3 HbF HbA2

(α2γ2) (α2δ2)
Separate and Glycate

HbA1c – C.E.
Homozygous β variant
Hb X
α2β*2
α α
X
Hb A
α2β2
β β
 
Hb A2
α α α2δ2
 
Hb  
F
α2γ
2

β globin chain variants in homozygous state
• One additional peak on

electrophoresis
(variant of Hb A)
• No Hb A
Absence • Centering on Hb F
No HbA = No HbA1c of Hb A
• If no zones: mix with
(α2β2) normal control

• In Summary
– Hgb E Homozygous
– No HbA0, therefore NO HbA1c
– Note: HbA1c is the Glycated Form of HbA0
– Must undergo Hb Electrophoresis Test, to rule out the
concern (Patient had Hemoglobin Disorder)
As per ADA and NGSP ……
– (Glucose Test) Fructosamine or Glycated Albumin - At
Least 1 month glycation of RBC
– FBS (initial phase of monitoring)
NGSP Guidelines
Homozygous Case
Alternative Glucose
Testing
Homozygous Hb E
No HbA = No HbA1c
No misleading HbA1c result by
CAPILLARY ELECTROPHORESIS
RESEARCH STUDIES AND JOURNAL
CONSISTENT
INCONSISTENCY
IFCC EXPERT
≤ 15%
Sebia Capillarys Flex Piercing Hb A1c method

registered by NGSP with no interferences of main
hemoglobin variants
PREVALENCE OF
HEMOGLOBINOPATHY in
The Philippines
DATA OF SCREENED HbP & THALASSEMIA BY
RUNNING HbA1c ELECTROPHORESIS
• PHILIPPINE GENERAL HOSPITAL, MANILA
– HbA1c Electrophoresis Test – 60 to 90 per day
– Screened HbP & Thal – 3 to 4 per day
– Prevalence of HbP & Thal – 4.4%
• MAKATI MEDICAL CENTER, MAKATI
– HbA1c Electrophoresis Test – 80 to 120 per day
– Screened HbP & Thal – 4 to 6 per day
– Prevalence of HbP & Thal – 5%
• Most common Hemoglobin disorders incidentally detected
during HbA1c testing by CE:
– HbE
– β-thalassemia
– α-thalassemia
• Abrupt Estimation (i.e) – Philippine Population closed to 120
million – est. % of HbP & Thal is around 5% equal to 6.5 million
population is either trait, carrier or homozygous.
(data with the permission of Makati Med. Center Laboratory Data worklist @ Dr. Del Valle & Philippine General Hospital
Laboratory main data extracted at phoresis – 2017) / No Exact Data Given for Confidentiality)
PHILIPPINE GENERAL HOSPITAL
HbA1c Electrophoresis Test & Screening of
Hemoglobinopathies & Thalassemia
• This clearly shows the
Prevalence of Hemoglobinopathies
& Thalassemia being screened
incidentally while running the
HbA1c Electrophoresis Test. This
will help the patient being informed
that he might be a carrier, trait or
asymptomatic.
- The electropherogram/curve
information shows the superiority
of Capillary Electrophoresis in
giving the adequate diagnosis for
HbA1c Test.
Philippine Population having
Hemoglobinopathy & Thalassemia is
around 15 % - 20%
By Average;
21.6 Million Filipino is having
Hemoglobinopathy & Thalassemia, and this is
the population having the risk or result
problem of HbA1c Test using another
technology
Source: Newborn Screening & Thalassemia Society Phils.

INTERESTING CASES:
From
HbA1C – CE (Capillary Electrophoresis
Technology)
Test Running
In
Vicente Sotto Memorial Medical

Center
Hgb E Heterozygous
Beta Thalassemia
Alpha Thalassemia with
Hgb H
Hgb O-Arab with Beta
Variant Hgb Constant Spring
Foetal Hemoglobin
Delta Thalassemia
Alpha Thalassemia Hgb E Heterozygous
Beta Thalassemia
Hgb Constant Spring
Intermedia
INTERESTING CASES:
From
HbA1C – CE (Capillary Electrophoresis
Technology) and Hemoglobin Electrophoresis
Test Running
In
Western Visayas Medical Center

HEMOGLOBIN E / BETA THALASSEMIA
NORMAL NEWBORN RESULT
HbA1c – C.E. (ALPHA THALASSEMIA WITH HGB H)
BETA THALASSEMIA MAJOR
HbA1c – C.E. (HGB E HETEROZYGOUS or Hgb AE)
HbA1c Standardisation
• NGSP : Started
HbA1c standardisation
in 1996
• IFCC : Developped a reference

system for HbA1c (IFCC-RM)
in 2004
• Master Equation
NGSP= (0.09148x IFCC)
+2.15

What is the Gold Standard
method for HbA1c
measurement?
Why is HPLC considered the Gold
Standard method for HbA1c
measurement?
The DCCT study
From 1983 until 1993, the DCCT (Diabetes Control and Complications Trial) was the
first clinical study to demonstrate the central role of HbA1c for the management of
diabetic patients. The Bio-Rad Biorex 70 HPLC method was used as the reference
anchor for the DCCT trial and this method was then adopted as the standard by the
NGSP (National Glycohemoglobin Standardization Program) in the US...
Bio-Rad DIAMAT HPLC

analyzer used with the
Biorex 70 kit
The Biorex 70 HPLC Reference Method
The IFCC (International Federation Clinical Chemistry) realized that this Biorex 70
HPLC method was not specific as the HbA1c fraction was co-eluting with other
substances which had the same elution behaviour as HbA1c but were not HbA1c by
definition...
? HbA0
Chromatogram obtained
HbA1c on the Biorex 70 HPLC
National Initiatives
Standardisation
Mono-S
NGSP JDS/
s i o n JSCC
n f u
Co
Comparison of National Reference Methods:

• Arbitrarily chosen
• Not specific
• Different numbers
• Obsolete (methods developed in the 70s)
Need to achieve a more uniform
standardization of HbA1c measurement
•To define what it is really HbA1c

•To have a reference method that measures only a well-defined analyte.
The New IFCC Reference Method
In 2002, to overcome the problem of lack of specificity of the HPLC, and in order
to standardize HbA1c testing worldwide, the IFCC developed two new
international reference methods that specifically measure the concentration of
HbA1c:
the LC/MS and LC/CE methods are today’s IFCC reference system for HbA1c and
they represent the only valid anchor to implement standardization of the
measurement
1. Digest, Glu-C 2. Separate, LC 3. Detect, MS

1 mg Hb Zorbax SB-CN, [M+2H]+2
pH 4.3, 37 °C, 18 hours 2.1 x 150 mm
50 °C, 300 l/min
IFCC LC/CE REFERENCE METHOD
1. Digest, Glu-C 2. Separate, LC 3. Collect, 4. Separate, CE

concentrate 5. Detect,
UV 214 nm
NOTE:
LC MS – LIQUID CHROMATOGRAPHY MASS SPECTROMETRY

LC CE – LIQUID CHROMATOGRAPHY CAPILLARY ELECTROPHORESIS
Primary reference materials IFCC Standardization
IFCC reference measurement procedures
LC/CE or LC/MS IFCC
Network
Secondary reference materials
Manufacturer’s internal
reference measurement procedure
Manufacturer’s
working calibrator Manufacturer
Manufacturer’s standing
measurement procedure
Manufacturer’s
product calibrator
Routine measurement procedure Individual

laboratory
Patient sample
Effects of the IFCC standardization of HbA1c
measurement
« HPLC is the Gold Standard! »
• This is a wrong idea
• HPLC was once the Gold Standard but Science evolves and there is now
an International reference measurement system within the concept of
metrological traceability: the IFCC Reference Method
INTERNATIONAL FEDERATION OF CLINICAL

CHEMISTRY (IFCC)
LC MS – LIQUID CHROMATOGRAPHY MASS SPECTROMETRY

LC CE – LIQUID CHROMATOGRAPHY CAPILLARY ELECTROPHORESIS
HbA1C Test – THE NEW GOLD STANDARD
IFCC REFERENCE METHOD

IZED !
CO G N
DE D & RE
MME N
RECO
Letter Confirmation from IFCC Expert
HbA1c by
capillary
electrophoresis
What is measured by CE
%HbA1c =
100 x [HbA1c area / (HbA1c area + HbA0 area)]
CE uses the same calculation

formula as the IFCC Reference
Methods
HPLC Calculation What is really being
measured?
the following calculation is applied:
%HbA1c =
HbA1c area / ∑ HbA area
[100 x (HbA1c area / (HbA1a1 + HbA1a2 +

HbA1b + HbF + Labile A1c + Carbamylated
Hb + HbA1c + HbA0 Gluthatione + HbA0
oxydated + HbA0 + HbA2) area) x slope +
Intercept] x EMG algorithm
Hb A0 Hb E, Hb D
HPLC
Long elution time
Just keep the essential
P4
Clear-cut separation
?
Hb A1c
A1a
A1b Unknown P3
? ?
? Hb F Unknown ?
? LA1c
? ?
Capillary
Electrophoresis:
The Next
Generation of
Separation
Method
SEBIA Capillary Electrophoresis Technologies
Your new solution
for
the analysis
of
Hb A1c Electrophoresis
(The Latest INNOVATIVE Technology for HbA1c Testing)
PRECISION
and
ACCURACY
cal&
n alyti ence
o u t A erfer
h t
Wit gical In
lo
B io
Treatment
and Therapy
of the Patient
How do we measure
A1c
Strict IFCC
formula
• The Hb A1c assay should be performed using a certified method (NGSP, DCCT, IFCC).
• Factors that can influence blood glucose levels, such as hemoglobinopathies, should be
taken into account when interpreting the Hb A1c value!
A1c Testing - Current
Methods
%GHb = Total GHb /

(Total non-GHb + Total GHb)
% HbA1c = HbA1c %HbA1c = [HbA1c] /

Area / ∑ HbA area [Total Hb]
%HbA1c = HbA1c / NEW REFERENCE GLOBAL /

(HbA1c + HbA0) GOLD STANDARD
LIQUID CHROMATOGRAPHY MASS SPEC – CAPILLARY ELECTROPHORESIS (LCMS-C.E.)
SEBIA
MACHINE
ARRAY
300 Samples/h
CAPILLARY
S RANGE
100 Samples/h
78 Samples/h
20 Samples/h
Use of capped whole blood samples.
Stored at 2-8°C during 7 days or Sample agitation
72 hours at room temperature or + piercing + hemolysis
3 months at -70 / -80°C
Migration
at 12.000 volts
Capillary in silicate and
Blue LED thermo-conductive resin
Detector (25µm diameter)
Direct
Injection < 1nL
quantification
at 415 nm
Cathode - Anode +
Special carousel
Use of capped whole blood
samples. Stored at 2-8°C
with tube guides
during 7 days or 18 hours at for hemoglobin
room temperature or analysis
3 months at -70 / -80°C Sample agitation
+ piercing + hemolysis
Migration
until 10.000 volts Capillary in silicate and
thermo-conductive resin
(25µm diameter)
Blue LED
Detector
Direct
Injection < 1nL
quantification
at 415 nm
Cathode - Anode +
ELECTROPHEROGRAM
CURVE / GRAPH
HbA0
Precise and Accurate Result

Without Any Analytical &
Straightforward Hgb Variant
Biological Interference
Detection
Indicator for Thalassemia
HbA1c
HbA1a+1b+3 HbA2
HbF
HbA1c Electrophoresis
Capillarys/Minicap flex piercing Hb A1c
with elevated Hb A1c
HbA0
HbA1c
HbA2
Profile in orange
Heterozygote Hb S variant
on Capillarys/Minicap FP HbA1c
Hb S0 (Sickle Cell)
Glycated Hb S
HbA1c
Hb A/S
No interference
with A0 and A1c peaks
Both NGSP and IFCC
Remarks and
Suggestion (manually
or programmable)
Methods Comparison
TOSOH BIORAD (HPLC) SEBIA
(HPLC) (CAPILLARY ELECTROPHORESIS)
HbC0
No direct interference on HbA1c
HbA0 HbC0
No alteration of the result
HbA0
HbA1c
HbA1c
HbC1c ???
HbC1c ???
Precision &
Accuracy of
capillary
electrophoresis
Quality Control in
the Laboratory
HbA1c: What is really being measured ?
Craig C. Foreback, Ph.D.
HbA1c in 2010, Part V - Westgard QC
When we talk about the quality of an assay for glycated

hemoglobin, we consider factors like imprecision, bias, total error,
and whether or not the method has the “seal of approval” of the
NGSP certified to provide equivalent results. If the statistics look
good and it is certified by the NGSP, what else do we need to know?
No one wants to talk about what we are really measuring.

Diagnostic companies label their method as an assay for
Hemoglobin A1c (HbA1c).
But is that what is really being measured?

PRECISION: Definition
The International Vocabulary of Metrology defines the

Precision as the closeness of agreement between
indications or measured quantity values obtained by
replicate measurements on the same or similar objects
under specified conditions [ISO/IEC Guide 99:2007].
PRECISION study
Precision was tested using the CLSI protocol EP5 [ 5 ] and measured for 20
working days, 2 runs a day, with a duplicate per run, and with 10 other samples
included in the run) at low (38 mmol/mol) and high (68 mmol/mol) HbA 1c
levels.
HbA1c: Performance of the SEBIA CAPILLARYS 2 Flex Piercing

C. Weykamp, H. Waenink-Wieggers, E. Kemna, C. Siebelder
Clin Chem Lab Med (2012); DOI: 10.1515/cclm-2012-0560
ACCURACY: Definition
Although the good precision of an analytical measurement is

necessary for the reliability of a result, it is not a sufficient
condition to guarantee the validity of the result.
• Any method showing the best precision with the lowest CVs
cannot, based on this only distinction, claim reliable results in
all circumstances. The accuracy, which is defined as the
closeness of agreement between a measured quantity value
and a true quantity value of a measurand [ISO/IEC Guide
99:2007], goes beyond as it refers to a combination of
Trueness and Precision [ISO 5725-1:1994].
Poor trueness Poor trueness
Poor precision
Not accurate Good precision
Not accurate
Good trueness Good trueness

Poor precision Not accurate Good precision Accurate
For internal use only - Not for customers
distribution
For internal use only - Not for customers
distribution
For internal use only - Not for customer
distribution
TOSOH
BIO-RAD
ROCHE
SEBIA
For internal use only - Not for customer

distribution
For internal use only-Not for customer
217
distribution
Risk Analysis: (Based on CLSI Protocol Guidelines & NGSP)
Results: trueness, precision and linearity - TEa
source: clinical chemistry journal: endocrinology and metabolism
HPLC
Let’s take the example of Variant II Turbo with a TEa specification of 6%

“If 100 patient samples were tested between QC events, 2.96%
of results would be unreliable while in-control, and in the worst
case, as many as 11 unreliable patient results (out of 100 results
total) could be expected when an out-of-control event occurs”
distribution
Let’s take the example of Tosoh G8 with a TEa specification of 6%

“If 100 patient samples were tested between QC events, 8 % of
results would be unreliable while in-control, and in the worst
distribution
IMMUNOASSAY
Let’s take the example of Integra 800 with a TEa specification of 6%

“If 100 patient samples were tested between QC events, 39 % of
results would be unreliable while in-control, and in the worst
distribution
Capillarys Electrophoresis is the Most Reliable Method
• “In contrast, the Capillarys 2 Flex Piercing assay had imprecision

and bias profiles that gave the highest patient weighted at each TEa
tested. Only 0.02% of results are predicted to be unreliable while
in-control, and < 1 unreliable patient result (out of 100 results total)
would be expected even for the worst case out-of-control
condition” For internal use only-Not for customer
distribution
CAPILLARYS

distribution
IFCC CERTIFICATION
NGSP CERTIFICATION
3 Key Sebia Hba1c message!
1. Superior
Analytical
performances
2. Free 3. Unique
Analytical medical added
interferences value

Take Home Message:

• WHY IT IS IMPORTANT THAT YOU
HAVE A MACHINE IN THE
LABORATORY THAT COULD DETECT
PRECISE HBA1C RESULT AND COULD
DETECT HEMOGLOBINOPATHY AND
SEPARATE FROM HbA1C RESULT?
• 1. To Avoid Falsely Elevated or inconsistent HbA1c

Result
• 2. To Know if the Patient is having Hemoglobin Disorder
(remember Hgb Variant could affect HbA1c Result)
• 3. Physiopathological condition of the RBC could affect
the proper treatment of the Clinician to the Patient. (Its
important to know that HbA1c is having a precise result)
INTERPRETATION WISE:
HbA1c Electrophoresis = Screening for HbP & Thalassemia
Hemoglobin Electrophoresis = Confirmatory
HbA1c – C.E. Hgb Electrophoresis
Zoning
Display
Pure Hemoglobin
QUANTIFICATION OF Hbg VARIANT

MARKETING WISE:
HbA1c Electrophoresis = Screening for HbP & Thalassemia

Hemoglobin Electrophoresis = Confirmatory
• Hematologist, Hemato-Pathologist,
Pathologist or Clinicians had its own client
for HbE Test & SPE Test
• But eventually The HbA1c Electrophoresis
Test will help the Laboratory/Hospital
Market the Hemoglobin Electrophoresis
Test
In Summary…
Warning!!! Pathological Condition which might affect red blood cell life span
Sebia Hb A1c:
deserving your diabetic patients
• Hb A1c by Sebia CE is measured according to the IFCC formula
• Sebia CE is accurate and shows excellent analytical

performances (low bias, low CV)
• Sebia CE does not suffer from any analytical interferences
• Sebia CE is able to detect the most common

hemoglobinopathies and quantify Hb A2, without interference
on Hb A1c measurement
• Sebia CE brings medical added values necessary for a correct

Hb A1c interpretation
SEBIA Hemoglobin Capillary Electrophoresis...
The most comprehensive method for initial screening
• Easy detection of the most common Hb variants
 Full separation of Hb S, C, D and E from Hb A, A2 and F
• Reliable screening of Beta-thalassemias
 Accurate quantification of Hb A2 and Hb F,
with no interference from Hb adducts
• Straightforward detection of Alpha-thalassemias
 Clear separation and quantification of Hb H
and Hb Bart’s
• Fully automated multi-parameter instruments
 Hb on adult and newborn, Hb A1c,
serum and urine proteins, CDT
• High resolution at a high throughput
 Up to 38 tests per hour (CAPILLARYS 2 Flex Piercing)
• Fast validation process using Mosaic screen
 Automatic presumptive identification and quantification
INSTALLATION
of
CAPILLARY
ELECTROPHORESIS in
the PHILIPPINES
(using HbA1c Electrophoresis & Special Test)
TOP GOVERNMENT and MEDICAL
CENTERS
• PGH MAIN – CAP 3 OCTA and MINICAP FLEX
• PGH MEDICAL RESEARCH CENTER (MRL) – MINICAP FLEX
• NATIONAL KIDNEY and TRANSPLANT INSTITUTE - CAP 3 OCTA and
MINICAP FLEX
• SAN LAZARO HOSPITAL – MINICAP FLEX
• JOSE REYES MEMORIAL MEDICAL CENTER – CAP2FLEX
• LUNG CENTER HOSPITAL – MINICAP FLEX
• PHILIPPINE CHILDREN AND MEDICAL CENTER (PCMC) – MINICAP
FLEX
• VICENTE SOTTO MEMORIAL MED. CENTER (CEBU CITY) –
CAP2FLEX & CAP 3 OCTA
• WESTERN VISAYAS MEDICAL CENTER – CAP 3 OCTA
• NORTHERN MINDANAO MEDICAL CENTER – CAP 3 OCTA
• EASTERN VISAYAS MEDICAL CENTER – CAP 3 OCTA
TOP GOVERNMENT and MEDICAL
CENTERS
• U.P. NATIONAL INSTITUTE OF HEALTH - CAPILLARYS 2

• DR. JOSE FABELLA MEMORIAL MEDICAL CENTER – CAP 3OCTA
• ILOCOS TRAINING and REGIONAL MEDICAL CENTER – MINICAP
FLEX (LA UNION)
• MARIANO MARCOS MEMORIAL MEDICAL CENTER (ILOCOS NORTE)
– MINICAP FLEX
• BATANGAS MEDICAL CENTER – MINICAP FLEX
• CAGAYAN VALLEY MEDICAL CENTER – MINICAP FLEX
• BATAAN GENERAL HOSPITAL & MEDICAL CENTER – MINICAP FLEX
LIST OF TOP PRIVATE HOSPITALS
1. MAKATI MEDICAL CENTER CAP 2 FLEX & HYDRASYS

2. HI PRECISION LABORATORY CAP 2 FLEX PIERCING
3. THE MEDICAL CITY (ORTIGAS) CAP 3 OCTA & MINICAP FLEX
5. UST HOSPITAL MINICAP
6. ST. LUKES HOSPITAL (QC) MINICAP FLEX
7. ST. LUKES HOSPITAL (GLOBAL) MINICAP FLEX / HYDRASYS
8. MANILA DOCTORS HOSPITAL CAP 3 OCTA
9. ASIATIC LABORATORIES CAP 2 FLEX PIERCING
10. RIVERSIDE MEDICAL CENTER CAP 2 FLEX PIERCING
11. LIPA MEDIX MEDICAL CENTER MINICAP FLEX PIERCING
12. UNIVERSITY OF CEBU MEDICAL CENTER MINICAP FLEX
13. CEBU DOCTORS UNIVERSITY HOSPITAL CAP 2 FLEX PIERCING
14. PERPETUAL HELP HOSPITAL, CEBU MINICAP FLEX
NEWLY INSTALLED (2015 - 2016)
HOSPITAL MACHINE
1. MANILA DOCTORS HOSPITAL CAPILLARYS 2 FLEX

2. CHINESE GENERAL HOSPITAL MINICAP FLEX
3. ILOCOS TRAINING & REG. MED. CENTER MINICAP FLEX
4. SUPERCARE MED. SERVICE INC. – MANILA CAP2FLEX
5. SUPERCARE MED. SERVICE INC. – MAKATI MINICAPFLEX
6. SUPERCARE MED. SERVICE INC. – ILOILO MINICAP FLEX
7. CALAMBA MEDICAL CENTER MINICAP FLEX
8. LUNG CENTER OF THE PHILS. MINICAP FLEX
9. CIUDAD ZAMBOANGA HOSPITAL CAPILLARYS 2 FLEX
10. DLSU MEDICAL CENTER CAPILLARYS 2 FEX
NEWLY INSTALLED (2017)
HOSPITAL MACHINE
1. MARY MEDIATRIX MEDICAL CENTER MINICAP FLEX
2. PHILIPPINE CHILDREN MEDICAL CENTER MINICAP FLEX
3. MARIANO MARCOS MED. CENTER MINICAP FLEX
4. ST. LOUIS UNIVERSITY HOSPITAL MINICAP FLEX
5. MCU MEDICAL CENTER MINICAP FLEX
6. MEDICAL CENTER MANILA CAP2FLEX PIERCING & MINICAP

FLEX
7. SAN PABLO MEDICAL CENTER CAP2FLEX PIERCING
8. ST. LUKES M ED. CENTER – QC MINICAP FLEX (UPGRADE)

HOSPITAL MACHINE
1. CDO MEDICAL CENTER CAP 2 FLEX
2. VICENTE SOTTO MEMORIAL MEDICAL CENTER CAP 2 FLEX & CAP 3

OCTA
3. BATANGAS MEDICAL CENTER MINICAP FLEX
4. METRO ANTIPOLO HOSPITAL & MED. CENTER MINICAP FLEX
5. URDANETA SACRED HEART HOSPITAL MINICAP FLEX
6. ACCUSERVE LABORATORY CAP2FLEX PIERCING
7. CGH, SAN PABLO LAGUNA MINICAP FLEX

HOSPITAL MACHINE
1. RAMIRO COMMUNITY HOSPITAL, TAGBILARAN CITY

MINICAP FLEX
2. COMMUNITY GENERAL HOSPITAL, SAN PABLO CITY MINICAP FLEX
3. MARITIME LABORATORY, MANILA CITY CAP 2 FLEX
4. ST. LUKES MEDICAL CENTER – GLOBAL MINICAP FLEX &

HYDRASYS
5. PASIG CITY GENERAL HOSPITAL , PASIG CITY MINICAP FLEX
6. BATAAN GENERAL HOSPITAL, BATAAN CITY MINICAP FLEX
7. ANTIPOLO HEALTH SYSTEM, ANTIPOLO CITY MINICAP FLEX
8. HOLY CHILD HOSPITAL, DUMAGUETE CITY CAP 2 FLEX

NEWLY INSTALLED (2020 -2021)
HOSPITAL MACHINE
1. OUR LADY OF CALCUTTA MEDICAL CENTER MINICAP FLEX
2. BINANGONAN LAKEVIEW HOSPITAL MINICAP FLEX
3. RPHS BINANGONAN MINICAP FLEX
4. GAT ANDRES MEMORIAL HOSPITAL MINICAP FLEX
5. BATANGAS MEDICAL CENTER MINICAP FLEX
6. NATIONAL KIDNEY AND TRANSPLANT INSTITTE CAP 3 OCTA
9. THE MEDICAL CITY – ORTIGAS CAP 3 OCTA
10. OSPITAL NG MANILA MINICAP FLEX

NEWLY INSTALLED (2020 -2021)
HOSPITAL MACHINE
1. THE MEDICAL CITY – ORTIGAS MINICAP FLEX
2. ST. FRANCIS MEDICAL CENTER, CDO CITY MINICAP FLEX
3. DON EMILIO MEMORIAL MEDICAL CENTERMINICAP FLEX
4. PERPETUAL HELP HOSPITAL, LAS PINAS MINICAP FLEX
5. CIUDAD MEDICAL ZAMBOANGA MINICAP FLEX
6. PHILIPPINE GENERAL HOSPITAL MINICAP FLEX /

FLC TEST
7. GALLARES MEMORIAL MEDICAL CENTER CAP 3 OCTA
8. WESTERN VISAYAS MEDICAL CENTER MINICAP FLEX

(JANUARY
2022)
9. VICENTE SOTTO MEMORIAL MEDICAL CENTER CAP 3 OCTA
HOSPITAL MACHINE
1. CAPITOL UNIVERSITY MEDICAL CENTER CAP 3 OCTA
2. THE MEDICAL CENTER CLARK MINICAP FLEX
3. SILIMAN UNIVERSITY MEDICAL CENTER CAP 3 OCTA
4. PHILIPPINE GENERAL HOSPITAL CAP 3 OCTA (UPGRADE)
5. OUR LADY OF PILLAR HOSPITAL MINICAP FLEX
6. NORTHERN MINDANAO MEDICAL CENTER CAP 3 OCTA
7. PHILIPPINE GENERAL HOSPITAL CAP 3 OCTA (UPGRADE)

HOSPITAL MACHINE
1. EASTERN VISAYAS MEDICAL CENTER CAP 3 OCTA
2. BUKIDNON PROVINCIAL MEDICAL CENTER MINICAP FLEX
3. ST. ELIZABETH MEDICAL CENTER CAP 3 OCTA
4. ADVENTIST MEDICAL CENTER (CEBU) MINICAP FLEX

UPCOMING INSTALLATION (2023)
HOSPITAL MACHINE
1. EAST AVENUE MEDICAL CENTER CAP 3 OCTA
2. PHILIPPINE HEART CENTER MINICAP FLEX
3. TONDO MEDICAL CENTER CAP 3 OCTA
4. QUIRINO MEMORIAL MEDICAL CENTER CAP 3 OCTA
5. VETERANS MEMORIAL MEDICAL CENTER CAP 3 OCTA
6. DAVAO DOCTORS HOSPITAL CAP 3 OCTA & MINICAP FLEX

ACCURATE & PRECISE RESULT
WITHOUT ANY ANALYTICAL & HbA1c Testing
BIOLOGICAL INTERFERENCE (HbA1c – CE)
LABORATORY SOLUTION
BENCE JONES PROTEIN

& KIDNEY LESION
TYPING
(UPE)
VERSATILE INNOVATIVE
TECHNOLOGY
HEMOGLOBINOPATHIES AND CHRONIC ALCOHOL ABUS

THALASSEMIA (CDT)
(HBE)
MULTIPLE MYELOMA / MYELOMA CONFIRMATION
MONOCLONAL (IT)
GAMMOPATHIES (SPE)
Boronate Ion-exchange Capillary
immunoassays
affinity HPLC Electrophoresis
Robustness Evolution of the Methods
Separation
‘To God be the Glory Always ’

UNIVERSITY of SAN AGUSTIN - Capillary Electrophoresis Lecture 2023

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UNIVERSITY of SAN AGUSTIN - Capillary Electrophoresis Lecture 2023

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WILHELM C.

CAPILLARY ELECTROPHORESIS TECHNOLOGY:

• Capillary Electrophoresis Test and Application

MASS SPECTROMETRY TECHNOLOGY

PCR (POLYMERASE CHAIN REACTION)

STRICTLY CONFIDENTIALTO SEBIA - NOT

• Capillary electrophoresis is an analytical technique that separates

Electro-endosmotic flow Electrical field

The Electro-Osmotic Flow (EOF) is a stronger force than the Electrical

HbA1c Screening and

1. Serum Protein Electrophoresis (SPE)

STRICTLY CONFIDENTIALTO SEBIA - NOT

Acute inflammatory pattern and

Screening Peak/BJ typing Precision medicine

STRICTLY CONFIDENTIALTO SEBIA - NOT

Analysis within 24 hrs on Minicap

- Detection of monoclonal protein on electrophoresis with specific antisera

STRICTLY CONFIDENTIALTO SEBIA - NOT

IgA Kappa IgD Kappa

IgM Kappa IgE Kappa

IgG Lambda IgM Lambda IgE Lambda

Abnormal peak The peak disappears in Ig G

Conclusion: Detection of monoclonal Ig G Kappa

– Hemoglobinopathies (Sickle Cell, Hgb Variants) & Thalassemia's (Alpha or Beta).

STRICTLY CONFIDENTIALTO SEBIA - NOT

Births with a pathological hemoglobin disorder (per 1,000 live births)

Immigration Trends Impacting Thalassemia

Alpha Thalassemia with Hgb H

WHO Resolution on Sickle Cell

WHO Resolution on thalassemiaand

UNESCO Resolution on Sickle Cell

UN Recognition of Sickle Cell as a pu

 1. G6PD DEFICIENCY 3 – 5 years as per

- Hb 11.5 g/L (115 – 145)

Anemia, Microcytosis, Hypochromia

Hgb Electrophoresis Fraction

The presence of Hb H, reduced HbA2,

Hgb Electrophoresis Test

Ferritin Result – 139.7ng / Ml

DETECTION OF DIFFERENT HGB VARIANT

Migration zones of 525 Hb variants in the database

• Worldwide population: 7,5 billions approx.

We always refer to the diabetes epidemy...

Data from WHO and

• Worldwide population: 7,5 billions approx.

... while we are still considering

Criteria for the diagnosis of Diabetes

Fasting plasma glucose FPG ≥ 126 mg/dL

 Hb A1c: biomarker of Diabetes

 Hb A1c: measure of hemoglobin A glycation

 Hb A1c reflects mean glycemia for the previous 3 months

 Hb A1c provides clinicians information about a long-term glycemic

HbA1c is currently defined as:

All of these are HbA1c

Glycohemoglobin, or GHb, or Total GHb, is defined as:

All of these are GHb (but not HbA1c)

 Hb A1c: initially used only for the monitoring of Diabetes

 New guidelines that promoted Hb A1c as a diagnostic test: