Smallest minimum weight according FI V and USP 3940
Smallest net weight w
according USP: 38 2015; 392016
Sr =
Sr = repeatability n = 10 = not more than 0. 10
W =
Sr = repeatability n = 10 = not more than 0. 10 d = scale interval
For analytical balance d = 0.1 or 0.01mg ; micro-balance 0.001 mg, w will be = 82 mg , 8.2 mg
, 0.82 mg
if SD = 0 or , 0.41 d Or w = 2000 SD
USP 35FI V : Tolerance not exceed 0. 1 of the amount weight; so smallest weight must be 100.0 4 digitsand 10.00 mg 5 digits, or
1.000 mg 6 digits.
Data below, usually are valid only for new balance
Also USP 38 2015 and USP 39 2016
SelectivitySpecificity
= the most important parameter
SpecificitySelectivity
USP, ICH, FDA: Specificity AOAC, IUPAC : Selectivity
Specificity is the ability to determine
the analyte unambiguously in the presence of other components, this includes degradation
products, metabolites, matrix component etc.
products, metabolites, matrix component etc.
Selectivity means that the method provides responses for a number of chemical entities that may
distinguished from each
others. It means also the ability to separate
the target analyte from all other components
Selectivity evaluation for pharmaceutical preparations:
Identity of API should be tested, and its selectivity should be proved.
Please be careful if the identification method that described in Pharmacopeia Official method is not identical with its
in Pharmacopeia Official method is not identical with its assay method e.g. Identification: IR, color reactions; assay :
HPLC
Why for some cases, identification of API in pharmaceutical preparations should be used by using 1 methods: IR, TLC, Color reactions
“IR Library” should be validated routine
A B
C R DQC Lab. at Pharmaceutical Industry
Nice if related
compounds, possible
degradation products,
Densitograms λ λ
λ
λ = 260 nm obtained from: 1 solution of standard mometasone furoate, 2 extract from excipients of laboratory-made cream, 3 extract of laboratory-made cream, 4 solution of
nipagin, 5 solution of nipasol, 6 extract of commercial ointment-1, 7 extract of commercial lotion, 8 extract of commercial cream-1, 9 extract of commercial ointment-2 and 10 extract of commercial
cream-2. Peak identities: A mometasone furoate, B nipagin and nipasol, C unknown.
Wulandari, L, Tan, KS., Indrayanto,G. 2003, J. Liq. Chromatogr. R T, 26, 109-117 A
Standard products,
known impurities
can be tested
The target peak must be proved regarding its identity and purity
Peaks should be well separated
F.Melianita, J. Witha, S.Arifin, WF.Karina, G. Indrayanto 2009. J. Liq. Chromatogr. R T, 32, 567-577
Rs should be not less 1.75
according USP Medicines Compendium, 2013
IF Impurities, related compounds, possible
Degradation products
were unavailable
Cited from: D. Widiretani, I. Luailia, G. Indrayanto,
J. Planar Chromatogr. 262013 37-42
If we have no impurities andor
degradation products;
we should do stressed
forced experiments to prove the selectivity.
Identity and purity check
of peaks A and B must be performed
Cited from: D. Widiretani, I. Luailia, G. Indrayanto,
J. Planar Chromatogr. 262013 , 37-42
Determination of Xanthorrhizol in the rhizomes of Curcuma xanthorrhiza by GC FID
Xanthorrhizol ?
F. Melinata G. Indrayanto 2006, Unpublished work
Should tried 3 different conditions
For FID, RID, ELSD = Blind methods
Purity Check using PDA detector HPLC or Densitometer TLC-Scanner
Cited from: M. Yuwono G. Indrayanto, Profile Drugs Excipients and Related Methodology, Vol. 32, Elsevier, 2005.
M.W. Dong, Modern HPLC for Practicing Scientist, Wiley, 2006
The purity of the peak can be calculated by
comparing the MS of the u, a and d of the TIC peak.
Acetylsalicylic acid
200 nm N=12804, Tf=1.20
Evaluation of the shape of acetylsalicylic acid Evaluation of the shape of acetylsalicylic acid
chromatogram using 5 wavelengths chromatogram using 5 wavelengths
0 5 10 15min
200 nm
220 nm
240 nm
260 nm
280 nm N=12804, Tf=1.20
N=12970, Tf=1.20
N=12924, Tf=1.20
N=12887, Tf=1.20
N=12924, Tf=1.20
Analysis using LC MSMS triple Quad
Cited from Agilent LCMSD triple Quad training, Singapore, May 2007
TA Sasaki, Chrom. Tech. JulyAugust 2008
Commission Decision 2002657EC
Accoding to CD 2002657EC Off.J.Eur.Commun. 2002,L221,8
CD 2002657EC Off.J.Eur.Commun. 2002,L221,8
Mw: 868.04
M. Distl et al. 2009 Potato Research 52, 39-56 Mw:
852.07
The power of MRM
MRM is nicer compare to SIR or SIM
Why we need do the “ Why we need do the “identity test
identity test” and “ ” and “
purity test purity test” of the peaks for
” of the peaks for each each samples for
samples for quantitative analysis ?
quantitative analysis ?
•If the peak is not the target compound, the conclusion will be not correct.
•If “impurity components” are included in the target peak, the quantitative-results
in the target peak, the quantitative-results will be wrong
•By Using LC-MSMS or GC-MSMS the target peak doesn’t need to be pure, it can
contain 1 compounds which have same target
ion; the Identity of the peaks can be differentiated by using
MSMS spectrum but ratio of qualifier to quantifier ions should be in the acceptable range
Linearity Calibration
Linearity
•
The linearity of a method is its ability to provide measurement results that
are directly proportional to the concentration of the analyte , or are directly
proportional after mathematical transformation.
• The linearity is usually documented as the
ordinary least squares OLS
curve, or simply as linear regression curve,
of the measured responses peak area or height as a function of increasing analyte concentrations
• Peak area
shall be preferred compared to peak heights
for making the calibration curve
• The linearity of
the detector can be obtained by diluting of the analyte stock solution,
whilst the linearity of the analytical method
can be determined by making a series of concentration of the analyte from
independent preparations weighing, spiking
G. Indrayanto, Suciati, M. Yuwono 2010, in Encyclopedia Chromatography 3
rd
Edition,Taylor Francis, pp.2336-2339