9/8/2019

CÁC PHƯƠNG PHÁP

PHÂN TÍCH CÔNG CỤ

TS. PHAN THỊ ANH ĐÀO

SÁCH, GIÁO TRÌNH CHÍNH:

1. Nguyễn Thị Thu Vân, Phân tích
định lượng, NXB ĐHQG Tp.HCM,
2009. (Thư viện).
2. Nguyễn Thị Thu Vân, Bài tâp và
Sổ tay Phân tích Định lượng,
NXB ĐHQG Tp.HCM, 2012 (Thư
viện).
SÁCH THAM KHẢO:
3. John Kenkel, Analytical Chemistry
for technicians, CPR Press, Taylor &
Francis Group, London (page:LMS).
4. Hồ Viết Quý, Các phương pháp
phân tích công cụ trong hóa học hiện
đại, NXB Đại học Sư phạm, 2005
(Thư viện).

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ASSESSMENTS
 Hình thức thi: Trắc nghiệm + Tự luận
 Giữa kỳ (50%) bao gồm:
1. Bài kiểm tra 1: (trắc nghiệm + tự luận) 50/3 %
2. Bài kiểm tra 2: (trắc nghiệm + tự luận) 50/3 %
3. Kiểm tra 3: …… 50/3 %
4. Điểm cộng (0,5-1,0 đ): hoạt động trên lớp
Cuối kỳ (50%): Trắc nghiệm + Tự luận (75-90 phút)

CHƯƠNG 1: GIỚI THIỆU 4 tiết

1.1. Giới thiệu
1.2. Đối tượng, nhiệm vụ, nguyên tắc, phân loại của
các phương pháp phân tích công cụ
1.3. Các phép đo lường cơ bản
1.4. Chuẩn bị mẫu
1.5 Mẫu trắng và mẫu đối chứng
1.6 Các phương pháp định lượng mẫu

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PHÂN LOẠI CÁC PP PHÂN TÍCH

HÓA PT 2
HÓA PT 1

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R=KC

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Steps characteristic of instrumental analysis methodology

CALIBRATION – HIỆU CHUẨN

Calibration is a procedure by which any instrument or
measuring device is tested with a standard in order to
determine its response for an analyte in a sample for which
the true response is either already known or needs to be
established.
a) The true response is already known
- Calibrating a balance: some known weight
- Calibrating a pH meter: the pH of a buffer solution

b) True response is established

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THE CALIBRATION OF AN INSTRUMENT

- a single standard: resulting in a calibration constant

- a series of standards, resulting in a standard curve

R: the instrument readout

R=KC C: the concentration od the analyte
K: proportionality constant

UNKNOWN SAMPLE SOLUTION

EXTERAL CALIBRATION- NGOẠI CHUẨN

PP ĐƯỜNG CHUẨN

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A STANDARD CURVE

Khoảng tuyến tính

EXAMPLE 1

SOLUTION

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A STANDARD CURVE
Finding the “Best” Straight Line y  mx  b

x = independent variable
y = dependent variable
m = slope
b = y-intercept

where:
y
slope  m
x

FINDING THE “BEST” STRAIGHT LINE

Determining the Best fit to the Experimental Data:
Method of Linear Least Squares is used to determine the
best values for “m” (slope) and “b” (y-intercept) given a set
of x and y values

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FINDING THE “BEST” STRAIGHT LINE

Goodness of the Fit: Dựa vào Hệ số tương quan- Hệ số hồi
qui
R2: compares the sums of the variations for the y-values to the
best-fit line relative to the variations to a horizontal line.
A perfect fit has an R2 = 1; no relationship for R2 ≈ 0

R2=0.9952 R2=0.5298
Very weak to
no relationship

Strong direct
relationship

99.5% of the y-variation is due to the x-variation 53.0% of the y-variation is due to the x-variation
What is the other 47% caused by?

Ý nghĩa Giá trị: -1 ≤ r ≤ 1

• r = 1: sự tương quan chính xác giữa hai biến
• r = 0: có sự độc lập hoàn toàn giữa hai biến
• r = -1: sự phụ thuộc đối nghịch nhau giữa hai biến.
• 0,90 < r <0,95: đường chuẩn tương đối tốt;
• 0,95 < r < 0,99, đường chuẩn tốt;
• r > 0,99 đường chuẩn rất tốt

R phải đạt yêu cầu: 0,995 R  1

Hoặc: : 0,99 R2  1

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CALIBRATION CURVE: shows a response of an

analytical method to known quantities of
analyte
PROCEDURE:
a) Prepare known samples of analyte
covering convenient range of
concentrations.

b) Measure the response of the

analytical procedure.

c) Subtract average response of blank

(no analyte).

d) Make graph of corrected response

versus concentration.

e) Determine best straight line.

Impact of “Bad” Data Points

y=0.16x + 0.12 R2=0.53261 y=0.091x + 0.11 R2=0.99518

Remove “bad” point

Improve fit and

accuracy of m and b

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Determining Unknown Values from Calibration Curves

(i) Knowing the values of “m” and “b” allow the value of x to be
determined once the experimentally y value is known
(ii) Know the standard deviation of m & b, the uncertainty of the
determined x-value can also be calculated

Limitations in a Calibration Curve

Limited application of calibration curve to determine an
unknown.
- Limited to linear range of curve
- Limited to range of experimentally determined response
for known analyte concentrations
Uncertainty increases further
from experimental points

Unreliable determination
of analyte concentration

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V0

Vs

STANDARD ADDITION- PP THÊM CHUẨN

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STANDARD ADDITION- PP THÊM CHUẨN

Concentration of analyte in initial solution signal from initial solution


Concentration of analyte plus s tan dard in final solution signal from final solution

X i 
IX
Standard addition equation:
S  f   X  f IS X

Total volume (V):

V  Vo  VS , Vo  unknown initial volume, VS  added volume of s tan dard

X  f   X i  Vo  S  f  S i  VS 
V  V 

X-intercept (y=0) yields

which is used to calculate  X i from:

 
X i  X  f  V
 Vo 

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SAMPLE

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HOME WORK : Tooth enamel consists mainly of the mineral

calcium hydroxyapatite, Ca10(PO4)6(OH)2. Trace elements in
teeth of archaeological specimens provide anthropologists with
clues about diet and disease of ancient people. Students at
Hamline University measured strontium in enamel from
extracted wisdom teeth by atomic absorption spectroscopy.
Solutions with a constant total volume of 10.0 mL contained
0.750 mg of dissolved tooth enamel plus variable
concentrations of added Sr. Find the concentration of Sr.

Added Sr (ng/mL = ppb) Signal (arbitrary units)

0 28.0
2.50 34.3
5.00 42.8
7.50 51.5
10.00 58.6

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70

60

50

0 28 40

2.5 34.3
5 42.8 30

7.5 51.5 20
y = 3.136x + 27.36
10 58.6 R² = 0.9973
10

0
0 2 4 6 8 10 12

[X]=8.72 ppb

INTERNAL STANDARD - PP NỘI CHUẨN

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INTERNAL STANDARD - PP NỘI CHUẨN

Known amount of a compound, different from

analyte, added to the unknown.
 Signal from unknown analyte is compared against signal
from internal standard
Relative signal intensity is proportional to
concentration of unknown
- Valuable for samples/instruments where response varies
between runs
- Calibration curves only accurate under conditions curve
obtained
- relative response between unknown and standard are
constant
 Widely used in chromatography
 Useful if sample is lost prior to analysis

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Area of analyte signal  area of s tan dard signal 

=K
 F  
Concentration of analyte  Concentration of s tan dard 

Area under curve

proportional to concentration
of unknown (X) and internal
standard (IS)

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HOME WORK: A solution containing 3.47 mM X

(analyte) and 1.72 mM S (standard) gave peak areas
of 3,473 and 10,222, respectively, in a
chromatographic analysis. Then 1.00 mL of 8.47 mM S
was added to 5.00 mL of unknown X, and the mixture
was diluted to 10.0 mL. The solution gave peak areas
of 5,428 and 4,431 for X and S, respectively

(a)Calculate the response factor for the analyte

(b) Find the concentration of S (mM) in the 10.0 mL of
mixed solution.
(c)Find the concentration of X (mM) in the 10.0 mL of
mixed solution.
(d)Find the concnetration of X in the original unknown.

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a)k=5,94
b)0,847 mM
c)6,163 mM
d)12,326 mM

The most common is through several dilutions of stock

standard solution: Serial dilution procedure

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DILUTION OF A STOCK STANDARD SOLUTION

DÃY CHUẨN- SERIAL DILUTION

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DÃY CHUẨN- SERIAL DILUTION

SAMPLE 1: A stock standard solution of
copper (1000.0 ppm) is available. Tell how you
would prepare 50.00 mL each of five standard
solutions, each required to be 1000.0 ppm,
and how you would proceed with the
preparation, including the kind of pipette
needed. The concentrations of the standards
should be 1.00, 2.00, 3.00, 4.00, and 5.00
ppm.

Definition: The blank is a solution that contains all the

substances present in the standards and the unknown (if
possible) except for analyte.
 The readout for such a solution should be zero, and as we have
indicated, the readout is often manually adjusted to read zero when
this blank is being measured. Thus, the blank is useful as a sort of
precalibration step for the instrument.

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Definition: A reagent blank consists of reagent

water and all reagents that normally are in contact
with a sample during the entire analytical procedure.
The reagent blank is used to determine the
contribution of the reagents and the preparative
analytical steps to error in the measurement.”
Another synonym for Reagent Blank is Method
Blank

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A control is a standard solution of the analyte prepared independently,

often by other laboratory personnel, for the purpose of cross-checking the
analyst’s work.
Thus, the analyst then knowns to scrutinize his or her work for the
puspose of discovering an error.

The results of the analysis of a control are often plotted on a control chart
in order to visualize the history of the analysis in the laboratory so that a
date and time can be identified as to when the problem was first detected.
Your instructor may wantto use controls and a control chart in various
experiments in this text.

THỬ HOẠT TÍNH DPPH

1,1-diphenyl-2-picryhydraxyl

54

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QUY TRÌNH THỬ HOẠT TÍNH DPPH

Control 100 ppm 50 ppm 25 ppm 10 ppm

Blank Sample Blank Sample Blank Sample Blank Sample Blank Sample

Mẫu 0 0 1200 600 600 300 300 150 120 60

EtOH 6000 1500 4800 900 5400 1200 5700 1350 5880 1440
DPP
H 0 1500 0 1500 0 1500 0 1500 0 1500

The concentration obtained from the standard curve is

rarely the final answer in a real-world instrumental
analysis.

- The sample must be diluted prior to the

measurement
- A chemical must be added prior to the measurement,
possibly changing the analyte’s concentration.
- The sample is a solid and must be dissolved or
extracted prior to the measurement.

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Sensitivity (độ nhạy) relates to the magnitude of change of a

measuring device (instrument) with changes in compound
concentration.

Limit of detection(LOD), in contrast to sensitivity, is

the lowest possible increment that we can detect with
some degree of confidence (or statistical
significance).
There are several ways to measure the LOD,
depending on the apparatus that is used.
The LOD often is reached when
the signal to noise ratio is 3 or
greater.

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According to the US Environmental Protection

Agency (EPA), the MDL is defined as “the
minimum concentration of a substance that
can be measured and reported with 99%
confidence that the analyte concentration is
greater than zero and is determined from
analysis of a sample in a given matrix
containing the analyte.

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DỰA THEO ĐỘ LẸCH CHUẨN MẪU

- Làm trên mẫu trắng (Reagent Balnk)
- Phân tích 10 lần song song (N 7), tính độ lệch chuẩn
(phải khác 0)

LOD= xmean + 3x s
- Nếu làm trên mẫu thử: 10 lần song song, nên chọn mẫu
thử có nồng độ thấp (khoang 5-7 lần LOD ước lượng)

LOD=3x s
* Đánh giá LOD theo R=mean/LOD
- 4<R<10: Nồng độ mẫu thử phù hợp
- R<4: dùng mẫu thử đậm đặc hơn
- R>10: Pha loãng mẫu thư

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DỰA THEO ĐƯỜNG CHUẨN

s: Sample Standard Deviation

m
hoặc a:
độ dộc
đường
chuẩn

DỰA THEO TÍN HIỄU NHIỄU (S/N)

LOD: S/N=3

LOQ: S/N=10

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