# R&S Compliance group, LLC. — Hazardous Materials Safety Interpretation - **operation:** document - **citation:** 14-0248 - **title:** R&S Compliance group, LLC. — Hazardous Materials Safety Interpretation - **source type:** guidance - **agency:** Pipeline and Hazardous Materials Safety Administration - **status:** guidance - **official:** true - **published on:** 2015-06-18 - **effective on:** Not available - **summary:** 14-0248 response to R&S Compliance group, LLC. concerning 172.101, 173.22, 173.50, 173.56. - **machine formats:** - **json:** https://regulus.evalyn.ai/document/phmsa-interpretation-14-0248.json - **markdown:** https://regulus.evalyn.ai/document/phmsa-interpretation-14-0248.md - **app url:** https://regulus.evalyn.ai/document/phmsa-interpretation-14-0248 - **source url:** https://www.phmsa.dot.gov/sites/phmsa.dot.gov/files/legacy/interpretations/Interpretation%20Files/2015/140248.pdf **body:** <<>> S. Departmen f Transportatio Washington. DC 20590 1200 New Jersey Avenue SE Pipeline and Hazardou: Administ titty JUN 18 2015 Mr. David Rivers R&S Compliance Group, LLC. 2707 W. Price Ave. Tampa, FL 33611 Reference No. 14-0248 Dear Mr. Rivers: This is in response to your December 30, 2014 letter requesting clarification of the Hazardous Materials Regulations (HMR; 49 CFR Parts 171-180) applicable to new explosives. You state your company receives a nitroglycerin and lactose mixture classified as "UN 3319, Nitroglycerin Mixture, Desensitized, Solid, N.O.S., 4.1" transported under approval EX2000120151. You further state that this raw material is used in the production of a pharmaceutical product. Various machines that come in contact with the raw material are cleaned and the cleaning process results in effluent waste material containing a methanol and water mixture with contaminate of nitroglycerin. You present in-house test data from tests run on several lots of effluent waste showing trace concentrations of nitroglycerin between 0.00616- 0.01220 mg/ml. You ask several questions regarding the need for classification of both effluent waste material and tablet nitroglycerin. Your questions are paraphrased and answered as follows: Q1. In order for § 173.56 to apply, does the resulting effluent material resulting from the alteration process (testing and cleaning) of the originally classified explosive mixture need to meet the definition of a Class 1 material in § 173.50(a) first? Or does § 173.56 apply first because the original ingredient that was altered was subject to § 173.56 and therefore all products or by-products of the initial classified explosive, regardless of the characteristics, or if the product meets the definition of Class 1 per § 173.50(a), require formal testing, classification and approval per § 107.705? Al. Per § 173.56(a) a new explosive is an explosive produced by a person who has not previously produced that explosive, or has previously produced that explosive but has made a change in the formulation, design, or process so as to alter any of the properties of the explosive. Any product containing an energetic ingredient (e.g., Nitroglycerin) is therefore subject to the requirements of § 173.56, and any modification of such product previously approved is also subject to the same requirements to determine if the modifications meet the definition of a new explosive. However, if the modification results in a new material that may be classified in a hazard class or division other than Class 1 under a proper shipping name specifically listed in the Hazardous Materials Table (HMT) in § 172.101 (e.g. UN1204), the modified material would not be subject to the provisions of §173.56 unless the proper shipping name has a special provision requiring approval under § 173.56. <<>> Q2. It the latter is true, does the final pharmaceutical product, nitroglycerin tablets, require the same approval to transport to pharmacies as the tablet is a result of an alteration of an initially classified explosive? A2. The nitroglycerin tablets would require approval in accordance with § 173.56(b). Q3. Would the resulting effluent material, containing residual amounts of what would be a classified explosive in other forms or concentrations, be properly described for transportation by other characteristics? In this case UN1204 Nitroglycerin solution in alcohol. A3. In accordance with § 173.22, it is the shipper's responsibility to properly classify a hazardous material. This Office generally does not perform this function. However, if the effluent waste can be appropriately described by an existing HMT entry for a hazard class or division other than Class 1 it may be oftered for transport under that entry. In your scenario, if the nitroglycerin solution in alcohol and water contains not more than 1 percent nitroglycerin in alcohol with no other energetic ingredients, the effluent would most appropriately be offered as UN1204. I trust this information is helpful. If you have further questions, please do not hesitate to contact this office. Sincerely, Duane A. Pfund International Standards Coordinator Standards and Rulemaking Division <<>> Webb 17356 Goodall, Shante CTR (PHMSA) New seposines From: Betts, Charles (PHMSA) 14-0248 Sent: Wednesday, December 31, 2014 12:05 PM To: Goodall, Shante CTR (PHMSA) Cc: Subject: Dodd, Alice (PHMSA); Foster, Glenn (PHMSA); Fink, William (PHMSA) Fw: Request for Interpretation Attachments: Request for interp less than 2 nitro.pdf; STM-AR-CV-0159 Nitroglycerin Cleaning Verification Method.pdf; STM-AR-FP-0164 Nitroglycerin Sublingual Tablets 0 3 mg 0 4 mg 06 mg.paf Shante - Please log and assign for response. Thanks From: Fink, William (PHMSA) Sent: Wednesday, December 31, 2014 09:08 AM Eastern Standard Time To: Betts, Charles (PHMSA); INFOCNTR (PHMSA) Subject: FW: Request for Interpretation Cc: Singh, Harpreet (PHMSA) Charles, R&S is requesting a letter of interp. Bill Fink From: Alston, Barbara CTR (PHMSA) Sent: Tuesday, December 30, 2014 4:43 PM To: Fink, William (PHMSA); Herrera, Jacqueline (PHMSA) Subject: FW: Request for Interpretation Good Afternoon Team, I spoke with Mr. Rivers on the phone and he has sent information pertaining to his questions/request, please see attachments. Many thanks, Barbara From: David Rivers [mailto:david@rscompliance.com] To: Alston, Barbara CTR (PHMSA) Sent: Tuesday, December 30, 2014 4:17 PM Subject: Request for Interpretation Barbara, Thank you for speaking with me today. The attached is a scenario requesting an interpretation as to the applicability of 173.56, "New Explosive" of an effluent waste from a pharmaceutical manufacturing and cleaning process. The narrative demonstrates the specifics of our situation. If needed, I also attached testing procedures that we use that ultimately creates the "by-product" we are trying to ensure we classify for transportation appropriately given the raw material Nitroglycerin. Your help is greatly appreciated. Happy New Year! David O. Rivers, M.S. 1 <<>> 2707 W. Price Ave., Tampa, FL 33611 Compliance Group, LIC Phone: (813) 433-4979 December 30, 2014 US Department of Transportation PHMSA - Energetic Materials Attn: Division of Approvals and Permit, PHH30 1200 New Jersey Ave. SE, East Building 2nd Floor Washington, DC 20590-0001 R&S Compliance Group, Ile would like to request a letter of interpretation as to the intent of the definition of a "New Explosive" described in 173.56. This section requires that a classification and new EX Approval be granted for the transportation of a classified explosive if the original classified explosive is altered "...as to change the formulation, design or process so as to alter the properties of the explosive." Specifically, we receive a Nitroglycerin and Lactose Mixture classified as; "Nitroglycerin Mixture, Desensitized, Solid, NOS with more than 2 percent but not more than 10 percent nitroglycerin, by mass", transported under Approval EX2000120151 and reclassified by the Associate Administrator as hazard class 4.1, UN3319. This raw material is used in the production of a pharmaceutical product, a Nitroglycerin Tablet. Each LOT of final product, the table themselves, must be tested for quality control according to Good Manufacturing Practices (GMP) and Food and Drug Administration (FDA) requirements. The machine used to extract the active ingredient from the raw material received, Nitroglycerin, also must be cleaned. Other machines which have contact surfaces that come in contact with the pharmaceutical mixture during the blending, mixing and tablet formation process must be washed. This testing and cleaning process results in effluent waste material containing a Methanol and Water Mixture with contaminates of Nitroglycerin. The in-house lab ran a test on this effluent waste material from several lots of effluent waste and found the following concentrations of trace Nitroglycerin per volume. (In house test procedures attached) Approximate Conc. of Nitroglycerin per container Container 1 0.01220 mg/mL Container 2 0.00068 mg/mL Container 3 0.00194 mg/mL Container 4 0.00616 mg/mL *Results are "Grab" samples from 55 gallon drums of effluent and not meant to meet the testing criteria prescribed in 173.52, 173.57 and 173.58 R&S Compliance Group, LLC <<>> ROSS 2707 W. Price Ave., Tampa, FL 33611 Phone: (813) 433-4979 Compliance Group, LIC Our question is if the requirements to seek proper classification and EX Approval from the Associate Administrator per 173.56 apply to the effluent waste from laboratory testing and/or cleaning processes by determining if these activities meets the definition of "altering" an existing explosive when the resulting effluent material to be transported, (transportation for disposal in this case), does not meet the definition of a Class 1 explosive ,173.50(a). #1 In order for 173.56 to apply, does the final effluent material resulting from the alteration processes (testing and cleaning) of the original classified explosive mixture (Nitroglycerin Mixture, Desensitized, Solid, NOS) need to meet the definition of a CLASS 1 material (173.50(a) FIRST? "an explosive means ..designed to function by explosion...",? If the resulting effluent material did in fact meet the explosive definition, we would agree an approval would we required as a New Explosive defined in 173.56. But what the resulting materials does not meet the definition of an explosive 173.50(a), would further examination of this effluent mixture and written determination be required from the Associate Administrator to state that the material does not need an approval? #2 Or, does 173.56 apply simply because the original ingredient that was altered was subject to 173.56 before initially offered for transportation, therefore, all products or by-products of the initial classified explosive (in this case, Nitroglycerin Mixture, Desensitized, Solid, NOS, EX2000120151), regardless of the resulting characteristics or meeting the definition of Class 1 set forth in 173.50 (a), require formal testing, classification and approval per 107.705. If the latter, #2, is true, does the final pharmaceutical product, the common Nitroglycerin tablet for heart patients require the same approval to transport to pharmacies as the tablet is the result of an alteration of an initially classified explosive? Or, does this resulting effluent material, even though it contains residual amounts of what otherwise would be a classified explosive in other forms or concentrations, simply need to be properly described for transportation to meet the definition of its characteristics, 173.22(a), not requiring an EX Approval, and in this case by 173.120(a)(2) as a flammable liquid such as; UN1204, Nitroglycerian Solution in Alcohol with not more than 1 percent nitroglycerin *This above authorized name does not list a special provision requiring an EX Approval from the Associate Administrator in order to describe a product as with the original Nitroglycerin product received and used, even though the description has the word "Nitroglycerin" within the name. This description would be selected by shippers where appropriate as required by "Shipper's Responsibility" R&S Compliance Group, LLC <<>> R8U 2707 W. Price Ave., Tampa, FL 33611 Phone: (813) 433-4979 Compliance Group, LIC We understand that there is not a threshold of an explosive concentration that PHMSA has established for an "altered" explosive, clarified according to interpretation #09-0058 which would need an EX Approval if, as stated in paragraph three, ... "new compositions containing any amount of explosive material...". This sentence refers to an "explosive" material presumably "explosive" by definition, 173.50(a). However, does the above described effluent waste, scenario, and furthermore the final pharmaceutical product, meet the intent or definition of a "New Explosive", subject to 173.56 simply because of "alteration" though a sampling and cleaning process and eventual formation and packaging or a pharmaceutical tablet involving previously "Classified Explosive" requiring an EX Approval for initial transport, where the "Process" is not intended to produce an a different "explosive", by definition, from the original "explosive" when the final effluent mixture does not exhibit characteristics of a CLASS 1 material, by definition? Your attention and interest to our request is appreciated. Please feel free to contact me with any questions you may have. Resiss CHMM, CSP, CET, CDGP David O. Rivers, M.S. Ph: 813-433-4979 R&S Compliance Group, lIc. Fx: 813-436-5392 www.rscompliance.com david@rscompliance.com R&S Compliance Group, LLC <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 1 of 32 Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 Test Method #: STM-AR-FP-0164 mg Effective Date: MAR 1 4 2014 Supersedes: 05/16/13 Prepared by: Date: 03/1/14 Reviewed by: Dsl Date: 03/1|/1L Reviewed by: Date: 03/11/14 Approved by: Ethe Bodes 03/12/14 Date: 13/2/14€ 03/15/14 I. PURPOSE The purpose of this procedure is to describe the test methods used for analysis of Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg and Intermediate products. TESTS A. Description B. Identification C. Assay D. Blend Uniformity . Uniformity of Dosage Units F. Degradation Products G. Dissolution H. Water Content I. Disintegration II. APPLICABILITY This method applies to Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg and Intermediate products. This method applies to Analytical Research Department. III. PROCEDURE EHS Safety Statements 1. Material Safety Data Sheets for all chemicals involved should be read prior to starting any chemical handling. All recommendations of work practices and Personal Protective Equipment (PPE) should be followed. Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 2 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg Test Method #:STM-AR-FP-0164 2. All solutions and prepared samples must be properly labeled, including hazard information, if they are to be stored longer than a single work shift. 3. All waste, including rinsings, should be labeled appropriately and handled per Watson Waste Management Policy, SOP GN-0026 and Risk Management Procedure RM-0018 (located on the Portal). If a hazardous material was used in a filter media, the filter media is considered hazardous material and must be disposed of appropriately. 4. All glassware should be inspected prior to use. Defective glassware should not be used. 5. Thick walled glassware should be used when vacuum degassing media and mobile phases. 6. Waste receptacles should be verified to have enough space to hold the volume of waste to be generated. 7. Flammable solvents should be dispensed inside of a chemical fume hood. Flammable solutions should be covered if stored outside of the hood. 8. Appropriate PPE should be worn based on the requirements of Risk Management Procedure RM-0009 (located on the Portal) and the MSDS. 9. When weighing powdered laboratory chemicals and products, excess materials should not be placed into the regular trash receptacles. Excess products should be placed inside a non-hazardous waste receptacle and excess chemical powders should be disposed of appropriately based on hazards present. 10. All PPE should be inspected prior to donning. PPE should be rechecked often for damage and replaced as needed. 11. Concentrated acids and bases should be handled with care and dispensed inside a chemical fume hood with appropriate PPE. 12. Incompatible waste streams should be segregated and collected separately for disposal. Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 3 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 Test Method #:STM-AR-FP-0164 mg A. Description Visually inspect the sample and compare with the description given in the respective Analytical research in-process and finished product specifications. Refer to the specification item number given in Section VI B. Identification 1. HPLC The major peak in the assay sample chromatogram corresponds to that of the retention time for the Nitroglycerin peak in the Standard chromatogram. 2. Thin-Layer Chromatographic Identification Test 2.1. Apparatus A chromatographic chamber made of inert, transparent material with flat bottom or twin trough, a tightly fitted lid and a size suitable for the plate is used. The chamber is lined on at least one wall with filter paper. Sufficient mobile phase is added to the chamber that, after impregnation of the filter paper, a depth appropriate to the dimensions of the plate used is available. The chamber is closed and allowed to equilibrate. 2.2. Reagents • Acetone, ACS grade or better • Ethyl acetate, ACS grade or better Toluene, ACS grade or better • Glacial acetic acid, ACS grade or better • Methanol, HPLC grade or better • Diphenylamine, ACS grade or better • Nitroglycerin standard 2.3. Reagent Preparation 2.3.1. Mobile Phase Mixture of toluene, ethyl acetate and glacial acetic acid in a ratio 16:4:1. Scale up or down as needed. Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> .... - Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 4 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 Test Method #:STM-AR-FP-0164 2.3.2. Spray Solution Prepare a mixture of diphenylamine in methanol (1 in 100). Scale up or down as needed. 2.4. Standard Preparation Prepare a solution of 1 mg of nitroglycerin in 1 mL of acetone. 2.5. Sample Preparation Transfer the amount of finely powdered Sublingual Tablets, equivalent to about 1 mg ot nitroglycerin, to a centrifuge tube, add 1 mL of acetone, cover with parafilm shake by mechanical means for 30 minutes, and Centrifuge at 3000 rpm for 10 minutes, use supernatant for analysis. 2.6. Procedure On a line parallel to and about 2 cm from the edge of a suitable thin-layer chromatographic plate coated with 0.25 mm layer of silica gel mixture: 1. Apply 10 uL of the test and standard solutions with an interval of at least 10 mm between the centers of spots. The application spots must be at 5 mm above the level of the mobile phase. 2. Allow the spots to dry 3. Place the plate in the chamber, ensuring the spots are above the surface of the mobile phase 4. Develop the chromatogram in a solvent system, until the solvent front has moved about three-fourth of the length of the plate. 5. Remove the plate from chamber, mark the solvent front and allow the solvent to evaporate. 2.7. Detection Spray with a solution of diphenylamine in methanol (1 in 100) and irradiate the plate with short (254 nm) and long (365 nm) UV light for about 10 minutes. Determine the chromatographic retardation factor (Rr) for the principal spots. The Rf of the principal spot obtained from the test solution corresponds to the obtained from the standard solution. Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 1 of 6 Test Method: STM-AR-CV-0159 Title: Nitroglycerin Cleaning Verification Method Effective Date: SEP 2 9 2011 Supersedes: 07/22/11 Prepared by: Date: 09/29/11 Reviewed by: Date: 09/24/11 Approved by: Date: 09/29/11 I. PURPOSE in manufacturing equipment. The purpose of this procedure is to describe the residue test method for monitoring Nitroglycerin II. APPLICABILITY This STM applies to cleaning swabs for Nitroglycerin. This method applies to both Analytical Research and Quality Control Departments. IlL. PROCEDURE A. Instrumentation High Performance Liquid Chromatography system consisting of a pump, an injector, a UV letector and a data processing system capable of integrating the areas under the curve of the peaki of interest. Column: Mobile Phase: Alltech, Alltima C18, 5 um, 4.6 mm × 150 mm PN# 88052 Injection Volume: USP Water: Methanol (60:40) 50 pL low rate un time 1.2 mL/min Not less than 20 min Wavelength: 220 (8) nm 25°C 10 min to 14 min B. Reagents 1. Water, USP 2. Methanol, HPLC grade or equivalent 3. Nitroglycerin standard C. Reagent Preparation 1 Mobile Phase: USP Water: Methanol (60:40) Mix 600 mL of USP water with 400 ml of Methanol and mix well. This preparation may be scaled up or down as needed Effective Date: 29 Sep 2011 11:24:09 GMT -06:00 USERS OF PRINTED DOCUMENTS ARE RESPONSIBLE FOR ENSURING THAT THE PRINTED DOCUMENT REPRESENTS THE MOST CURRENT VERSION Printed by: Production Activity Control (svs-aumas-001 Date/Time: 24 Dec 20111:03:16 PM GMT -06:00 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 2 of 6 STM#: STM-AR-CV-0159 Title: Nitroglycerin Cleaning Verification Method Effective Date: SEP 2 9 2011 Supersedes: 07/22/11 2 Diluent: USP Water: Methanol = 50:50 Mix 500 mL of USP water with 500 mL of Methanol and mix well. This preparation may be scaled up or down as needed D. Standard Preparation: 1. Stock Solution: Accurately weigh and transfer approximately 63.3 mg of Diluted Nitroglycerin Standard (equivalent to 0.6 mg of Nitroglycerin) into a 50 ml, volumetric flask, dissolve and dilute to volume with Methanol. The concentration of Nitroglycerin stock Solution is approximately 0.012 mg/mL. 2. Working Standard Solution Pipette 5.0 mL of the stock solution into a 50 mL volumetric flask, dilute to volume with diluent, and mix well. Scale up or down as needed. The stock and working standard solutions are stable for 5 days at room temperatures. Concentration of Nitroglycerin working standard is about 0.0012 mg/mL. E. Swab Sample Preparation: Swab samples are collected and submitted in 50-mL plastic centrifuge tubes. Pipette 5.0 mL of the diluent into each of the sample tubes and mix the contents for at least 2 minutes using a vortex mixer. Transfer about 1.5 mL of the resulting solution into an HPLC vial. Swab samples and solutions are stable for 3 days. F. Blank Swab Preparation: A Blank Swab is prepared and submitted in a 50 mL plastic centrifuge tubes. Treat the same way as a Sample Swab. G. HPLC Procedure: SiA, dad ip system according to te Make at neated mie " dumentien Pection System Suitability Test. Calculate the reproducibility (%RSD) for six Nitroglycerin Working Standard injections and perform the system suitability calculations. Calculate the reproducibility (ORSD is ≤ 5.0 %), the number of theoretical plates (N≥ 3000), and the tailing factor (T≤ 2.0). If the system is suitable, proceed with the samples analysis. Bracket the samples with the Working Standard Solution. Effective Date: 29 Sep 2011 11:24:09 GMT -06:00 Version: 2.0 USERS OF PRINTED DOCUMENTS ARE RESPONSIBLE FOR ENSURING THAT THE PRINTED DOCUMENT REPRESENTS THE MOST CURRENT VERSION. Printed by: Production Activity Control (svs-qumas-001) Date/Time: 24 Dec 20111:03:16 PM GMT -06:00 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 3 of 6 STM#: STM-AR-CV-0159 Title: Nitroglycerin Cleaning Verification Method Effective Date: SEP 2 9 2011 Supersedes: 07/22/11 H. Calculations Note: Label mobile phase, detergent and swab peaks in the sample chromatograms according to the table below (Refer to Validation Report ARVR-11-0068) Peak Peak RT (min) RRI Mobile Phase Peak 1 0.766 0.06 Detergent Peak 0.872 0.07 Mobile Phase Peak 2 1.274 0.11 Swab Peak 1 2.21 20.18 Mobile Phase Peak 3 3.368 0.28 Swab Peak 2 4.485 0.37. Swab Peak 3 6.232 0.52 Swab Peak 4 8.674 0.72 Nitroglycerin 12.084 1.00 Swab Peak 5 13.39 1.11 : Calculate the quantity (Q) of Nitroglycerin or unknown ug /swab using the following equation: ug Nitroglycerin /Swab = Au —X— WS XPFXO.1 As 0.661 Where PF = Potency of Nitroglycerin Reference Standard. Use as % (e.g. 0.948% is entered as 0.948) As = Average area of the Nitroglycerin peaks in chromatograms for the bracketing Working Standard Solutions. Aul = Peak response of Nitroglycerin in the sample chromatogram. Ws = Weight of Diluted Nitroglycerin Reference Standard in mg. 0.661 = Correction factor based on the ferrous plate recovery study, expressed in decimals. 0.1 = Dilution factor including standard, Sample dilutions and conversion of Diluted Nitroglycerin into Nitroglycerin Version: 2.0 Effective Date: 29 SeD 2011 11:24:09 GMT -06:00 USERS OF PRINTED DOCUMENTS ARE RESPONSIBLE FOR ENSURING THAT THE PRINTED DOCUMENT REPRESENTS THE MOST CURRENT VERSION. Printed by: Production Activity Control (svs-qumas-001) Date/Time: 24 Dec 20111:03:16 PM GMT -06:00 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 4 of 6 STM#: STM-AR-CV-0159 Title: Nitroglycerin Cleaning Verification Method Effective Date: SEP 2 9 2011 Supersedes: 07/22/11 IV. REFERENCES Current USP Current SOP # AR-0051, How to write a Standard Test Method. Cleaning Validation Request letter for Nitroglycerin (From Technical Service Department), received on 06/09/20011. V. HISTORY New, Effective date: 07/22/11 Revision 1, effective date: SEP 2 9 2011 Linearity range in the summary chart of page 6 updated to: 0.0001213 mg/mL to 0.002425 mg/mL VI. SPECIFICATION Cleaning verification criteria: NMT 5.77 ug of Nitroglycerin/ Swab Version: 2.0 Effective Date: 29 Sep 2011 11:24:09 GMT -06:00 USERS OF PRINTED DOCUMENTS ARE RESPONSIBLE FOR ENSURING THAT THE PRINTED DOCUMENT REPRESENTS THE MOST CURRENT VERSION Printed by: Production Activity Control (svs-qumas-001) Date/Time: 24 Dec 20111:03:16 PM GMT -06:00 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 5 of 6 STM#: STM-AR-CV-0159 Title: Nitroglycerin Cleaning Verification Method Effective Date: SEP 2 9 2011 Supersedes: 07/22/11 Figure 1. Chromatogram of Nitroglycerin Working Standard Solution SAMPLE INFORMATION Sample Name: Vial: Sample Type: Standard STD 1-7 Acqured By: 43 Acq. Method Set: Date Acquired: 7/11/2011 7:39:09 PM EDT atatkow! CV LC 17 : 50000 ul Percering tad: 6 - Santa et Nama: 37i0 tirely std stab 1C17 Toc. Chl. Desor.: DAD AUCh 1 Sample 220, Bw hennel Name A1100 DAD AU Ch1 00G- Auto-Scaled Chromatográm Mobile-Phase Peak 1 - 0.758 Mohile Phase Pook-2--1-272 Peak 32-8:369 2,000- Nilroglycering =12:070-- boblle Phale 5:00 2it 400 aco Minte 10/20 72.00 1400 Channel Name A1100 DAD AU CAT SampleName: STD 17 Simpletere Name 2 | STO 1-7 Molio Prace Feast 2589: SID 1-7 Dategent Fak 0759: 469 STD 1.7 STD 17 Sieb Poak1 Mobile Ptoe Feak2! 15744 Q.670| 400| _ 1.6 1272 3012 Motle Place Fears 35400 2210 1572 157.0 B5S 24 SID 17 Such Pa*2 4550 STD 1-7 Sian Rackt E410 B.870 Version: 2.0 Effective Date: 29 Sep 2011 11:24:09 GMT -06:00 201 1.2 G-DUMENTS ARE REne r. Production it or sums 0 CUMENT Rate me: 34 De 2011, 03-16 PM MIT-06.00 Printed bv: Production Activity Control (svs-qumas-001) <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 6 of 6 STM#: STM-AR-CV-0159 Title: Nitroglycerin Cleaning Verification Method Effective Date: SEP 2 9 2011 Supersedes: 07/22/11 Summary Chart Type of Swab: Texwipe TX714 A Maximum Allowable Limit: NMT 5.77 Mg / Swab Area to be Swabbed: 100 cm? Target Ingredient: Nitroglycerin Limit of Detection: 0.00006063 mg/mL Limit of Quantitation: 0.0001213 mg/mL Linearity: 0.0001213 mg/mL to 0.002425 mg/mL Specificity: No Interference Plate Recovery: From Ferrous Plates: 66.1 % Swabbing Solution: Methanol Swab Stability: Stable for 3 days Swab Stability in Solution: Stable for 3 days Swab Sampling Technique: As per SOP # AR-0063 Section: 8.6.1 Version: 2.0 Effective Date: 29 Sep 2011 11:24:09 GMT -06:00 USERS OF PRINTED DOCUMENTS ARE RESPONSIBLE FOR ENSURING THAT THE PRINTED DOCUMENT REPRESENTS THE MOST CURRENT VERSION. Printed bv: Production Activity Control (svs-aumas-001) Date/Time: 24 Dec 20111:03:16 PM GMT -06:00 <<>> ...-. Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 5 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg Test Method #:STM-AR-FP-0164 C. Assay 1. Instrument An HPLC system consisting of an autosampler, pump, a column, an adjustable wavelength UV Detector and an integrator capable of integrating the peaks of interest. HPLC Parameters: Column: PN# 00G-4252-E0 Phenomenex, Luna C18 (2), 5 um, 4.6 mm x 250 mm Mobile Phase: Methanol/Water=50/50 Run time: Injection Volume: 20 uL Flow rate: not less than 19 minutes Wavelength: 1.0 mL/min 220 (8) nm, ref: off Column Temperature: Rt window: 25 °C 13.5 minutes to 17.5 minutes 2. Reagents : Diluted Nitroglycerin Reference Standard Methanol, HPLC grade or equivalent USP Water 3. Reagent Preparation 3.1. Mobile Phase Preparation: Methanol/Water=50/50 Mix 1000 mL of Water with 1000 mL of Methanol and sonicate to degas for 5 minutes. This preparation may be scaled up or down as needed. Mobile Phase was demonstrated to be stable for at least 6 weeks at room temperature conditions 3.2. Diluent Use assay Mobile Phase: Methanol/Water=50/50 Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 6 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg Test Method #:STM-AR-FP-0164 4. Standard Preparation 4.1. Working standard solution Accurately weigh about 158 mg of Diluted Nitroglycerin standard equivalent do 15 re. or ito lycene dilent and mix wel 10 is pegition may be scaled up or down as needed. 4.2. Stability of Standard Solution The standard solution has been proved to be stable for 15 days at room temperature conditions. 4.3. Standard Concentration calculation The working solution contains approximately 0.075 mg/mL of Nitroglycerin | and is calculated using the following equation: Cstd =- Wstd× P 20 mLx100 Cstd Where, Concentration of Nitroglycerin standard in mg/mL. W std = P = Diluted Nitroglycerin Standard weight, mg Potency of the Diluted Nitroglycerin Reference Standard in %. 100% = Conversion from Diluted Nitroglycerin to Nitroglycerin 5. Sample Preparation 5.1. Stock Sample Solution Weigh the amount of tablets listed in the table below. Transfer into the proper volumetric flask. Add a stir bar and 80% of the flask volume of diluent. Stir the sample for 1 hour (tablets should completely disintegrate). Remove the stir bar and rinse it inside the flask with diluent, complete to volume with diluent and mix well. Transfer about 10 mL into a centrifuge tube, cover with paratilm and centrifuge the sample at 3000 rpm for 10 minutes. (Concentration is about 0.3 mg/mL). Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 7 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 Test Method #:STM-AR-FP-0164 mg Strength #of Tablets Volumetric Flask in mL 0.3 mg 20 20 0.4 mg 20 25 0.6 mg 25 50 5.2. Working Sample Solution Dilute 6.0 ml, of the centrifuged supernatant stock solution into a 25 ml. volumetric flask and dilute to volume with diluent, mix well. Transfer an aliquot into HPLC vial to analyze. (Concentration is in range 0.072 to 0.077 mg/mL). 5.3. Stability of Sample Solution least 3 days at room temperature conditions The Stock and working assay sample solutions were proved to be stable for at 6. Procedure 6.1. Refer to SOP AR-0049 "High Performance Liquid Chromatographic Analysis" 6.2. System Suitability Criteria Make five replicate injections of the Working Assay Standard-1 solution. Calculate the reproducibility (% RSD) of the five consecutive injections, theoretical plates (N) and the tailing factor (T) for the Nitroglycerin peak. The system is suitable if: RSD ≤ 2.0%, N≥ 3000 and T ≤ 2.5. Inject the Working Assay Standard Solution-2 twice. Compare the potency (%) of Standard-2 relative to Standard-1: 6.3. Standard Comparison Calculation Std Comp. (%) = - Astd2xWstd1 Astdl× Wsta2 -x 100 Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 8 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 | Test Method #:STM-AR-FP-0164 Astal Where, = Peak area of Standard-1 (Average of five injections) Asta2 = Peak Area of Standard-2 (Average of two injections) Wstal = Standard-1 weight, mg Wstd2 = Standard-2 weight, mg Standard-1 is suitable for use if Standard comparison (%) value is within the range 100.0 ‡ 2.0%. 6.4. Sample Analysis Inject working sample solutions bracketed by working standard -1. 6.5. Factors to be used in Empower Factor Description Custom Field in Standard Concentration Standard Concentration Empower Set up in mg/mL Component Editor Table/ 20 (for 0.3mg and STANDARDS ONLY 0.4mg) or 25 (for Number of Tablets for analysis Sample Weight 0.6mg) 83.3333 (for 0.3mg), 104.1667 (for Dilution volume in mi Dilution 0.4mg) or 208.3333 (for 0.6 mg) 0.3, 0.4 or 0.6 Nitroglycerin Strength Component Editor Table/ in mg STANDARDS & UNKNOWNS (under LC) 7. Calculations 7.1. Peak identification Follow table below for possible peaks observed in the assay sample chromatograms based on the specificity study during the method validation (Reference: 2492p9) Isers of printed documents are responsible for ensuring that the printed document represents the most current revisior Jocument No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 201‹ <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 9 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg Test Method #:STM-AR-FP-0164 Component name Peak Retention Time Placebo 2-Mononitroglycerin No peak observed (minutes) RRT NA Peak observed NA 2.964 1-Mononitroglycerin Peak observed 0.20 3.120 0.21 1,2-Dinitroglycerin 1,3-Dinitrolgycerin Peak observed Peak observed 5.240 0.36 5.838 Mobile Phase (Diluent) Peak observed 0.40 Nitroglycerin Peak observed 6.154 0.42 14.673 NA 7.2. Assay Calculation From the HPLC analysis, obtain the peak area of the sample (Asp) and Standard-1 (Asta). The percentage of Nitroglycerin per tablet is calculated from the equation: % LC =- AsplxCstdxVF×25mL Astd × LCx#tablets × 6.0mL - × 100% Where, Cstd = Aspl = Nitroglycerin Standard-1 concentration, mg /mL Astd Peak area of the sample. = LC = Average Area of Bracketing standard-1. #tablets = Label claim in mg number of tablets used in the analysis VFstock = Volume of the Stock Solution in mI D. Blend Uniformity 1. Instrument, Reagent, Reagent Preparation, Standard Preparation and Procedure Refer to Assay section III. C.1, III. C.2, III. C.3, III.C.4 & III.C.6. Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> -... Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 10 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 mg Test Method #:STM-AR-FP-0164 2. Sample Preparation Samples of one to six dosage units are provided from various blenders and drum locations. Perform the following for each sample: Tap the vial several times on the bench top. Weigh accurately the vial containing the sample (with the cap included in the initial weight) and record the weight (G). Quantitatively transfer the content of the vial to a volumetric flask (follow table below according strength and dosage units provided as a reference only). NOTE: Ensure to wash the vial and the inside portion of the cap with diluent Add a stir bar into the volumetric flask and diluent up to 70% of the flask volume and stir for 1 hour. Remove the stir bar and rinse it inside the flask with diluent. complete to volume with diluent and mix well. Transfer an aliquot into a centrifuge Tube, cover with pot of superman it it vial to anal pm for 10 minute. The Nitroglycerin concentration in the prepared Blend Uniformity Samples should be within the range of 0.045 mg/mL to 0.1125 mg/ml Dosage Units Strengths VFstock in mg in mL 0.3 5 1 dosage unit 0.4 5 0.6 0.3 10 2 dosage units 0.4 10 0.6 0.3 20 10 3 dosage units 0.4 0.6 20 25 Rinse the empty sample vials and caps with acetone and: -Dry the rinsed caps at room temperature for at least 2 hours or until needed -Dry the rinsed vials (without the cap) in the oven at 105°C for about 30 minutes, ensuring that they are completely dry Weigh the empty vial (with the cap) and record its weight (T). Finally calculate the weight of the sample (N) = (G-T). Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 11 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 Test Method #:STM-AR-FP-0164 mg 3. Procedure 3.1. Factors to be used in Empower Factor Description Custom Field in Standard Standard Concentration Empower Set up Concentration Component Editor Table/ Weight in mg/mL STANDARDS ONLY Sample Weight in mg Volumetric Sample Weight Volume Flask Dilution Factor in mL Dilution Theoretical potency Theoretical potency in Component Editor Table/ decimals (Not as in STANDARDS & percentage) UNKNOWNS (under LC) 4. Calculations The percentage of Nitroglycerin is calculated using the equation below: %LC=- Aspl Cstd xVF × 100% Astd Ws×%Th -x100% Where Aspl Astd = Peak area response of the sample. = Average area response of the bracketing standard. Cstd = Concentration of Nitroglycerin standard in mg/mL Ws = Sample weight in mg %Th = Theoretical percentage of the active in the blend. = Volume of the Volumetric Flask in mL E. Uniformity of Dosage Units (Content Uniformity) 1. Instrument, Reagent, Reagent Preparation, Standard Preparation and Procedure Refer to Assay section III.C.1, III. C.2, III. C.3, III. C.4 & III.C.6. Users of printed documents are responsible for ensuring that the printed document represents the most current revision. Document No.: STM-AR-FP-0164 Rev 10 Effective Date: March 14, 2014 <<>> Watson Laboratories, Inc.-Florida STANDARD TEST METHOD Page 12 of 32: Title: Nitroglycerin Sublingual Tablets, 0.3 mg, 0.4 mg & 0.6 Test Method #:STM-AR-FP-0164 mg 2. Sample Preparation Weigh ten tablets individually. Transfer into separate volumetric flasks (VF) follow table below). Add a stir bar and 70 % of the flask volume of diluent and sti for 1 hour. Remove the stir bar and rinse it inside the flask with diluent, complete t volume with diluent and mix well. Transfer an aliquot into a centrifuge tube, cover with parafilm and centrifuge the sample at 3000 rpm for 10 minutes. Transfer an Strength VF in mL 0.3 mg 5 0.4 mg 5 0.6 mg 10 Concentration of Nitroglycerin is from 0.06 mg/mL to 0.08 mg/mL 3. Procedure 3.1. Factors to be used in Empower Factor Description Custom Field in Standard Standard Concentration Empower Set up Concentration Component Editor Table/ 1 in mg/mL STANDARDS ONLY Number of Tablets for Sample Weight 5 (for 0.3mg and a - **truncated:** true - **body characters:** 74399