Good Weighing Practices At QC LaboratoryImpact of Weighing in Analysis * Weighing is a one of key activities in all the QC laboratories * Most of the time, our understanding is not at sufficient level + Its importance or complexity is underestimated. * Quality of weighing determines the Quality & Accuracy of final test result. * The USP specifically requires highly accurate results when weighing analytes for quantitative measures * Right choice of balances (Analytical/semi-micro/micro ) with desired resolution, accuracy & repeatability is essential to reduce the error and meet the complianceType of balances Balancename | Resolution decimal digits (gm) Ultra-microbalances 0.1 pg 0.0000001 Microbalances lug 0.000001 Semi-microbalances 0.01mg 0.00001 Analytical balances O.1mg 0.0001 Precision balances 1g +1mg ig+ imgBalance Selection for Minimum Weight Cire) (hate lame elereiel sii a mV I TU a ttt and Hy (S.D.) (typical) *3 0.001 mg 2.5 ug 3.0 mg 0.03 m, 20 m| 0.01 mg = : 0.02 mg 30 mg 0.1 mg 0.2 mg 120 mgWhat is minimum weight ? Minimum weight is the minimum sample quantity required to perform an accurate quantitative analysis is based on the measurement error of the balance used In order to satisfy the required weighing tolerance, when samples are weighed the amount of sample mass (i.e., the net weight) must be equal to or larger than the minimum weight. The minimum weight applies to the sample weight, not to the tare or gross weight. If the sample quantity is too small, the measurement error will be huge and result of the analysis will be unreliable.Requirement of Balances ‘SELECTIONCalculation of Minimum Mass Minimal mass value of balance can be established based on repeatability - analysis required precision 0,1%, - standard deviation (Balance specification) Eg Max 220g; d=0.01m@/0.1mg; 5 =0,025mg; k=2; A=0.1% © Max 220g; d=0.01mg/0,Img; s=0,025mg; k=2; A=0,1% “7 7. 0,025mg = 2000: 0,025mg = 50mg Minimal weighing for the balance is 50mg, if analysis is supposed to be performed with precision 0.1%.Definition: Accuracy * Closeness of agreement between a measured quantity value and a true quantity value of a measurand. VIM * * Difference between measurements average value and the real value according to USP Precision * Closeness of agreement between indications or measured quantity values obtained by replicate measurements on the same or similar objects under specified conditions. VIM Trueness closeness of agreement between the average of an infinite number of replicate measured quantity values and a reference quantity value (°) VIM-= International Vocabulary of Terms in Legal Metrology (*4) USP — United States PharmacopeiaAccuracy & Precision imprecisely, accurately (values are wide-spread) precise, accurately (good balance repeatability)USP General Chapters * Measurement is stated to be ‘accurately measured’ or ‘accurately weighed’, — (41) Balances — (1251) Weighing On An Analytical BalanceRequirement of Balances Select the appropriate balance based on the accuracy, repeatability, stability, access control, printout or connect to other instrument or LIMS etc. = URS * Select Correct accuracy and repeatability * Qualification * Installation at right place / location * Operation qualification * Performance qualification / Calibration = Password protection, Access control, printer, etcUnderstand the USP Requirement USP General Chapter <1251>; "In order to satisfy the required weighing tolerance, when samples are weighed, the amount of sample mass (i.e., the net weight) must be equal or larger than the minimum weight. The minimum weight applies to the sample weight, not to the tare or gross weight.Balances Requirement - USP Repeatability tolerance 0.10% 4 Expansion factor, K 2 Acceptance criteria 2x SD/N < 0.10% Smallest possible sample 820d (in optimal conditions) weight/lowest starting point Operating Range (2x SD x 1000) to max capacity or Tolerance Satisfactory if its weighing value is within 0.10% of the test weight value Test weight Between 5% and 100% of the balance's capacity Calibration uncertainty of the Must be < 1/3 of 0.10% weightLocation : Installation of Balances Install the balance on anti- vibration table & non-magnetic surface and grounded to prevent static electricity Room should be temperature and humidity controlled vibration, air currents, Should be free of drafts and away from air conditioner or fans or windows to avoid strong air current or direct sunlight Away from magnetic fields (magnetic stirrer), electromagnetic radiation eg RF generators/communication devices and electric motors, Away from Corrosive materials are used nearby.Operational Qualification OQ has to cover following, but not limited to; * Control of stable indication * Mechanical mobility of all moveable parts * Manually triggered or automatic adjustment by means of built-in weights. = Automatic adjustment reduce the drift of the balance. * Operation of ancillary equipment * Tare function * Calibration part of OQLinearity Eccentrici Performance Tests Ability of a balance to follow the linear relationship between a load and the indicated weighing value. Nonlinearity usually is expressed as the largest magnitude of any linearity deviation within the test interval. Deviation in the measurement value caused by eccentric loadiing—in other words, the asymmetrical placement of the center of gravity of the load relative to the load receiver, Eccentricity usually isexpressed as the largest. magnitude of any of the deviations between an off- center reading and the center reading fora given test load. NMT 0.05% deviation From 3 to 6 points where (41) is ‘over the range af the applicable. For other balance. uses, respective tolerance requirement divided by2. NMT 0.05% deviation Performed in the where (41) Is center of gravity &4 applicable. For other quadrants{ uses, respective Tet joad usually tolerance should be 303% of the requirement divided capacity of the by2. balance or higheric o “ Repeat-ability Performance Test Change in weighing value divided by the change in load, usually measured between zero and the capacity of the balance. Ability of a weighing instrument to display identical measurement values for repeated weighings of the same objects under the same conditions, €.g., the same measurement procedure, operator, measuring system, operating conditions, and same location over a short period of time, Repeatability usually is expressed as the standard deviation of multiple weighings. NMT 0.05% deviation Thetest load ator where (41) Is sufficiently close applicable. For other to the capacity of uses, respective the balance. tolerance requirement divided by 2. Requirement from (41) 10 replicate where applicable. For weighings (test other uses, user weight-few specified requirements percent of the will apply. nominal capacity ‘of the balance),Accuracy <41>: Accuracy: The accuracy is satisfactory if its weighing value, is within 0.10% of the test weight value. A test weight is suitable if it has a mass between 5% and 100% of the balance's capacity. maximum permissible error (mpe) or uncertinity shall 1/3 of the applied test ie 0.03%. (ASTM E617) Note: A readability of 0.1 mg of balance is believed as “accurate to 0.1 mg" as misconception.Balance Calibration According to USP General Chapter <41> “Balances”, for substances to be accurately weighed, the balance used must be calibrated over the operating range The most important are ; % Repeatability (RP), % Eccentricity (EC), %* Linearity (L) and Sensitivity (SE),Repeatability Why Repeatability is so important? It will have significant impact on all the quantitative analysis Ability of a weighing instrument to display identical measurement values for repeated weighings of the same objects under the same conditions, e.g., the same measurement procedure, same operator, same measuring system, same operating conditions, and same location over a short period of time. Systematic deviations normally can be prevented if Repeatability is performed.Repeatability test Perform 10 measurements with the same reference weight Calculate the Standard Deviation [SD] Repeatability is satisfactory if — 2x §D/Nominal reference weight used is < 0.10% Ifthe standard deviation obtained is less than 0.41d, (d-is the scale interval}, replace this standard deviation with 0.41d. — 2xSDx 1000 ifSD<0.41d, In this case, repeatability is satisfactory if two times 0.41d, divided by the nominal value of the weight used, does not exceed 0.10%. — 2xSD {0.41d) x 1000 = 820d — analytical balance with a readability of 0,1 mg, this means the starting point yielded is 82 mg.Repeatability : Repeatability : Individual measurement deviation from average value does not exceed standard deviation, that is ~ 0,0003g with probability 68,5%. Individual measurement deviation does not exceed three standard deviations, that is ~ 0,0009g with probability higher than 99,7%, so very close to certainty. - [Xe-9 a _ a Fe nce reper 1 Gimereaan sa hmeLinearity test * Linearity: To ensure that balance is accurate at the desired level in the Operating range * Ability of a balance to follow the linear relationship between a load and the indicated weighing value. * Nonlinearity usually is expressed as the largest magnitude of any linearity deviation within the test interval. * Perform 3 to 6 points over the range of the balance, = Limit: NMT 0.05% deviation where (41) is applicable. For other uses, respective tolerance requirement divided by 2. Bisa i Racer eters {tis a deviation of balance real curve from straight Jing joining two points A:B ~ ideal weight.Sensitivity Test Change in weighing value divided by the change in load, usually measured between zero and the capacity of the balance. Use certified weights with an appropriate weight class Perform as performed for repeatability test Limit: NMT 0.05% deviation where (41) is applicable. For other uses, respective tolerance requirement divided by 2.Eccentricity Test Deviation in the measurement value caused by eccentric loading— in other words, the asymmetrical placement of the center of gravity of the load Eccentricity usually is expressed as the largest magnitude of any of the deviations between an off-center reading and the center reading fora given test load. in practice, a difference is defined between indication when mass standard is put at central point of weighing pan and indication when the same mass. standard ts lacated at another place on the weighing pan. Performed in the center of gravity and the four quadrants Test load usually should be 30% of the capacity of the balance or higher. Limit: NMT 0.05% Deviation where (41) is applicable.Testing of Balance parameters How often balance parameters should be tested? Intervals defining balance calibration/testing shall be based on range of operation, their intensity, balance stability in time and expected weighing process measurement precision. Assuming that external conditions are stable, following balance parameters control periods can be fixed: — Calibration annually — Repeatability & centricity monthly — Sensitivity change weekly — Verificatian/ adjustment dailyFactors affects the weighing Accuracy There are several external factors influence the accuracy of weighing; * Ambient area and people influence result of weight measurement; * Major balance external factors are; — Oscillations, vibrations ~ Breeze of air — Temperature drifts — Electrostatics — Evaporation and absorption phenomena (hygroscopicity) — Magnetism * Other factors is nature of sample , * Both balance and the sample will influence the accuracy of results * To prevent any such issue , need to identify the reason properly — \sitdueto Balance —external factor or sample— external factor?Impact of Vibration & Strong Air * Oscillations — vibrations are transmitted by the ground and walls that are generates and affects weighing balance — Effect is longer measurement time and higher indication dispersion, — Prevent vibrations by keep away from vibration area — Keep on anti-vibration table. Anti vibration double rubber console, to suppress vibrations * Breeze of air influence to instability and long weighing time. — Balance workstation should not be located close to doors or windows. — Closeness to devices such as air-conditioning, fans, should be avoided,Volatile or hygroscopic samples Sample nature influence the weighing accuracy: © Volatile liquids (low boiling solvents or solid with volatile solvents ) can undergo evaporation during weighing © Balance indicates fluctuation / drift; ie weighing continuously decrease while measurement © Use appropriate weighing vessels, like bulbs with narrow necks or vessels with top cover. Weighing of Volatile samples: QO Weighing of low boiling liquid point in a vessel with a gas-tight enclosure of small diameter. O Close immediately after transfer of material QO After the balance display stabilizes, the analyst records the specimen weightImpact due to Temperature How Temperature affect the weighing; Weighing room temperature should be maintained at constant level. Eg variation must not more than 0.5°C/hour. Equilibrate the sample to room temperature before weighing, will give error due to heat convection ( hot sample will be less than true value) Precaution/ aspects for weighing process: = pick up the samples with use of tweezers or other holders = should not put their hands into weighing chamber = Touched with hands, samples —may change their temperatureoe & Hygroscopic samples Sample nature influence the weighing accuracy: Hygroscopic samples absorbs the moisture from ambient air and steadily gain the weight Measured weight will have higher mass than actual Weighed promptly Use the hermetic vessels or a gas-tight enclosure. Weighing vessel should be clean & dry and easily transferable Add the desired amount of sample, and replace the enclosureCorrosive and Bio-hazardous Samples Aseptic or Biohazardous Samples + Weighing the samples in the confines area / bio-safety cabinet /isolator, or similar containment device. * Care should be taken if airflow in the hood may cause balance instability Corrosive Materials * Extra care is essential when materials of this nature are weighed. * Use sealed containers such as weighing bottles or syringesSamples with Electrostatics Effect of electrostatic presence. © Slow drift of weighing result, © Large dispersion of weighing results ina series of measurements, and © No return to zero if a load is taken off the weighing pan. Possible source: — Dry, finely divided powders may be charged with static electricity — The static charge may develop due to low relative humidity, clothing worn and gloves used How to prevent: = Antistatic weigh boats, antistatic guns, and antistatic screens = Placing the container in a metal holder = Balances with a built-in antistatic device is available (piezoelectric components or low amount of a radioactive polonium) to generate a stream of ions that dissipateMagnetism as interfering factor If magnetic load is measured, electromagnetic field of a balance is disturbed or weighed sample is influenced by magnet installed in a balance. It will lead to incorrect mass reading of a weighed sample. High resolution balances are constructed on basis of electromagnetic sets which include a force-motor and magnet. How to avoid: Increasing a distance between a sample and balance mechanism. Use under-hook weighing with application of special racks or hooks made of aluminum.Precaution while weighing All receivers must be clean, dry, and inert. weighing uncertainty for small samples, i.e., net weights with a mass determined around repeatability. Receivers should be nonmagnetic used at ambient temperature Weighing dishes should be polymer or aluminum. Antistatic and compatible with the liquid sample. Safety measures: Use proper PPEs gloves, mask, goggles etc during a weighing to avoid exposure Hazardous materials should be handled in an enclosure that has appropriate air filtration. Many toxic— and possibly allergenic— substances are present as liquids or finely divided particles.