SEND Example.In vitro Neutral Red Cytotoxicity Assay

This example shows a report table, trial design and results data for the in vitro Neutral Red Uptake Cytotoxicity Assay in example study 123.

The following shows the data as it is represented in a report table for study 123. This report includes summary statistics (e.g. Average, Standard Deviation, Coeff. Var.) for each plate and an EC50 value for the entire assay. The grey cells in plate 1 are cells that do not contain the CHO Hamster cells.

Neutral Red Cytotoxicity Assay Results (Relative Absorbance Data)
	Sample ID	Smoke Regime	Replicate Number	Plate Number	Well Number		Relative Assay Plate Absorbance Readings
							Cigarette Smoke Condensate (µg/mL)								SLS (µg/mL)			EC50 (µg/ml)
						0	10	50	75	100	120	140	160	200	110	200	0
	030001	Medium	1	1	1	0.046*	0.046*	0.039*	0.012*	0.030*	0.003*	0.057*	0.032*	0.003*	0.050*	0.044*	0.040*
	030001	Medium	1	1	2	0.048*	107	77.7	65.7	30.2	15.6	9.79	5.41	2.49	8.09	0.791	0.014*
	030001	Medium	1	1	3	0.005*	98.6	77.4	57.7	35.8	8.09	7.36	3.22	0.791	6.38	0.061	0.050*
	030001	Medium	1	1	4	0.049*	94.9	81.5	38.2	31.2	7.60	5.17	3.22	0.791	5.17	0.304	0.017*
	030001	Medium	1	1	5	0.047*	111	78.1	64.5	32.9	8.82	5.90	2.74	1.28	10.3	0.547	0.011*
	030001	Medium	1	1	6	0.032*	96.6	76.2	48.0	26.8	7.84	4.20	2.74	1.52	8.09	4.93	0.022*
	030001	Medium	1	1	7	0.044*	105	77.9	58.2	28.3	8.09	4.68	1.76	0.547	8.57	1.76	0.056*
	030001	Medium	1	1	8	0.056*	0.013*	0.040*	0.008*	0.013*	0.002*	0.029*	0.018*	0.006*	0.016*	0.024*	0.046*
Average							101	78.2	54.5	29.9	9.70	6.17	3.37	1.19	8.94	1.22
Std. Dev.							6	2.9	9.4	3.6	2.95	2.19	1.12	0.72	2.72	1.58
Coeff. Var.							5.8	3.6	17.2	11.9	30.4	35.5	33.1	61.0	30.5	129.9
*media without cells
	030001	Medium	1	2	1		93.4	74.0	56.0	35.1	11.8	8.70	5.10	1.98	5.34	1.02
	030001	Medium	1	2	2		102	75.2	53.3	35.1	11.8	7.74	5.58	3.66	5.34	1.02
	030001	Medium	1	2	3		107	79.0	55.0	26.5	10.1	6.06	3.66	1.74	7.02	0.780
	030001	Medium	1	2	4		104	80.2	55.3	29.8	8.94	3.90	3.66	1.50	6.30	0.780
	030001	Medium	1	2	5		101	75.7	54.1	26.7	4.86	4.38	1.26	1.02	6.30	1.02
	030001	Medium	1	2	6		110	85.7	67.7	26.0	5.34	4.62	3.42	1.50	6.30	0.540
	030001	Medium	1	2	7		104	77.8	64.9	27.7	8.22	3.18	2.46	1.02	6.30	0.780
	030001	Medium	1	2	8		103	71.6	58.9	25.5	9.42	4.14	3.90	1.26	4.38	0.780
Average							103	77.4	58.1	29.0	8.82	5.34	3.63	1.71	5.91	0.840
Std. Dev.							5	4.4	5.3	4.0	2.63	1.97	1.37	0.86	0.83	0.170
Coeff. Var.							4.6	5.6	9.2	13.6	29.8	36.9	37.6	50.0	14.1	20.2
EC50 (µg/ml). Effective concentration at 50% cytotoxicity. (using a specific sigmoidal equation = model). (could also be EC20 or EC30, EC70, etc.)																		123

Assumption: A Trial (study) can have more than one assay type. (this line will be moved to the ts.xpt assumptions page)

Rows 1-2:	Show two records for TSPARMCD = "GLPTYP", using TSSEQ to indicate multiple records, since both GLP Types apply for this example study.
Row 8:	Shows that the sponsor's study reference ID is not applicable.
Rows 9-12:	Show that TSGRPID has been used to link records (name, location, country) related to the test facility (TSGRPID = 1). The Study Director is associated with the Test Facility.
Rows 18-24:	Show that this example study includes two different species of bacteria and a total of five strains.
Rows 18-22:	Show that TSGRPID (TSGRPID = 2) has been used to link the information for one species (Salmonella) with four different strains that are tested in this example study.
Rows 23-24:	Show that TSGRPID (TSGRPID = 3) has been used to link the information for one species (Escherichia coli) with the strain that is tested in this example study.

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Row	STUDYID	ASSAYID	DOMAIN	TSSEQ	TSGRPID	TSPARMCD	TSPARM	TSVAL	TSVALNF
1	123	NRU	TS	1		GLPTYP	Good Laboratory Practice Type	FDA
2	123	NRU	TS	2		GLPTYP	Good Laboratory Practice Type	OECD
3	123	NRU	TS	1		STSTDTC	Study Start Date	2022-05-25
4	123	NRU	TS	1		STITLE	Study Title	Determination of the in vitro genotoxicity potential using the in vitro Neutral Red Uptake assay
5	123	NRU	TS	1		SNDIGVER	SEND Implementation Guide Version	TOBACCO IMPLEMENTATION GUIDE VERSION 1.0
6	123	NRU	TS	1		SNDCTVER	SEND Controlled Terminology Version	SEND Terminology 2021-09-30
7	123	NRU	TS	1		SSPONSOR	Sponsor Organization	Example Sponsor Inc.
8	123	NRU	TS	1		SPREFID	Sponsor Study Reference ID		NOT APPLICABLE
9	123	NRU	TS	1	1	TSTFNAM	Test Facility Name	Example Test Lab Name
10	123	NRU	TS	1	1	TSTFLOC	Test Facility Location	10 Somewhere Street, Montgomery, AL 10000
11	123	NRU	TS	1	1	TFCNTRY	Test Facility Country	USA
12	123	NRU	TS	1	1	STDIR	Study Director	Dr. R. Smith
13	123	NRU	TS	1		GLPFL	GLP Flag	Y
14	123	NRU	TS	1		ASTD	Assay Standard	NIH Publication No. 07-4519
15	123	NRU	TS	1		ASTDV	Assay Standard Version	2006-11
16	123	NRU	TS	1		SSTYP	Study Type	GENOTOXICITY IN VITRO
17	123	NRU	TS	1		SSSTYP	Study Sub Type	In Vitro Neutral Red Uptake
18	123	NRU	TS	1		SPECIES	Species	Hamster
19	123	NRU	TS	1		TESTSYS	Test System	CHO

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Rows 1-20:	Show the test conditions for the set with a short name or SETCD value of CSC-50b. The sponsor chose this value to represent plate number 1, well number 1, with a concentration of 50 µg/mL of Cigarette Smoke Condensate as shown in the report table for study 123.
Row 17:	Shows the value for STRAIN/SUBSTRAIN is NO STRAIN because the well is a media only well, having no CHO Hamster cells.
Rows 21-40:	Show the test conditions for the set with a short name or SETCD value of CSC-50. The sponsor chose this value to represent plate number 1, well number 2, with a concentration of 50 µg/mL of Cigarette Smoke Condensate as shown in the report table for study 123.
Rows 41-60:	Show the test conditions for the set with a short name or SETCD value of CSC-50. The sponsor chose this value to represent both wells numbered 7 in plate number 1 and 2, with a concentration of 110 µg/mL of Sodium Laurel Sulfate as shown in the report table for study 123

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Row	STUDYID	ASSAYID	DOMAIN	SETCD	SET	TXSEQ	TXPARMCD	TXPARM	TXVAL
1	123	NRU	TX	CSC-50b	CSC-50-blank	1	TRTDMIN	Treatment Duration Minimum	47.5
2	123	NRU	TX	CSC-50b	CSC-50-blank	2	TRTDTRG	Treatment Duration Target	48
3	123	NRU	TX	CSC-50b	CSC-50-blank	3	TRTDMAX	Treatment Duration Maximum	48.5
4	123	NRU	TX	CSC-50b	CSC-50-blank	4	TRTDU	Treatment Duration Unit	HOURS
5	123	NRU	TX	CSC-50b	CSC-50-blank	5	INCBTMP	Incubation Temperature	37
6	123	NRU	TX	CSC-50b	CSC-50-blank	6	INCBTMPU	Incubation Temperature Unit	C
7	123	NRU	TX	CSC-50b	CSC-50-blank	7	WAVELGTH	Spectrophotometer WAVELENGTH IN NANOMETERS	540
8	123	NRU	TX	CSC-50b	CSC-50-blank	8	HUMID	Atmospheric Relative Humidity Percent	90
9	123	NRU	TX	CSC-50b	CSC-50-blank	9	ATMCO2	Atmospheric CO2 Percent	5
10	123	NRU	TX	CSC-50b	CSC-50-blank	10	SPTOBID	Sponsor defined tobacco identifier	CIG01a
11	123	NRU	TX	CSC-50b	CSC-50-blank	11	EXPTYP	Exposure Type	Air Liquid Interface
12	123	NRU	TX	CSC-50b	CSC-50-blank	12	SAMTYP	Sample Type	Whole Smoke Conditioned Media
13	123	NRU	TX	CSC-50b	CSC-50-blank	13	INTRVN	Name of the Intervention Article	Tobacco ProdA
14	123	NRU	TX	CSC-50b	CSC-50-blank	14	ITVTYPE	type of intervention article	PRODUCT
15	123	NRU	TX	CSC-50b	CSC-50-blank	15	ITVCONC	intervention concentration	50
16	123	NRU	TX	CSC-50b	CSC-50-blank	16	ITVCONCU	intervention unit	ug/ml
17	123	NRU	TX	CSC-50b	CSC-50-blank	17	STRAIN	Strain/Substrain	NO STRAIN
18	123	NRU	TX	CSC-50b	CSC-50-blank	18	SPDEVID	Sponsor defined device identifier	PUFFMASTER3k
19	123	NRU	TX	CSC-50b	CSC-50-blank	19	DUREFID	Smoking Regime	Medium Intensity Regimen
20	123	NRU	TX	CSC-50b	CSC-50-blank	20	SPTOBID	Sponsor defined tobacco identifier	CIG01a
21	123	NRU	TX	CSC-50	CSC-50	1	TRTDMIN	Treatment Duration Minimum	47.5
22	123	NRU	TX	CSC-50	CSC-50	2	TRTDTRG	Treatment Duration Target	48
23	123	NRU	TX	CSC-50	CSC-50	3	TRTDMAX	Treatment Duration Maximum	48.5
24	123	NRU	TX	CSC-50	CSC-50	4	TRTDU	Treatment Duration Unit	HOURS
25	123	NRU	TX	CSC-50	CSC-50	5	INCBTMP	Incubation Temperature	37
26	123	NRU	TX	CSC-50	CSC-50	6	INCBTMPU	Incubation Temperature Unit	C
27	123	NRU	TX	CSC-50	CSC-50	7	WAVELGTH	Spectrophotometer WAVELENGTH IN NANOMETERS	540
28	123	NRU	TX	CSC-50	CSC-50	8	HUMID	Atmospheric Relative Humidity Percent	90
29	123	NRU	TX	CSC-50	CSC-50	9	ATMCO2	Atmospheric CO2 Percent	5
30	123	NRU	TX	CSC-50	CSC-50	10	SPTOBID	Sponsor defined tobacco identifier	CIG01a
31	123	NRU	TX	CSC-50	CSC-50	11	EXPTYP	Exposure Type	Air Liquid Interface
32	123	NRU	TX	CSC-50	CSC-50	12	SAMTYP	Sample Type	Whole Smoke Conditioned Media
33	123	NRU	TX	CSC-50	CSC-50	13	INTRVN	Name of the Intervention Article	Tobacco ProdA
34	123	NRU	TX	CSC-50	CSC-50	14	ITVTYPE	type of intervention article	PRODUCT
35	123	NRU	TX	CSC-50	CSC-50	15	ITVCONC	intervention concentration	50
36	123	NRU	TX	CSC-50	CSC-50	16	ITVCONCU	intervention unit	ug/ml
37	123	NRU	TX	CSC-50	CSC-50	17	STRAIN	Strain/Substrain	CHO
38	123	NRU	TX	CSC-50	CSC-50	18	SPDEVID	Sponsor defined device identifier	PUFFMASTER3k
39	123	NRU	TX	CSC-50	CSC-50	19	DUREFID	Smoking Regime	Medium Intensity Regimen
40	123	NRU	TX	CSC-50	CSC-50	20	SPTOBID	Sponsor defined tobacco identifier	CIG01a
41	123	NRU	TX	SLS-110	SLS-110	1	TRTDMIN	Treatment Duration Minimum	47.5
42	123	NRU	TX	SLS-110	SLS-110	2	TRTDTRG	Treatment Duration Target	48
43	123	NRU	TX	SLS-110	SLS-110	3	TRTDMAX	Treatment Duration Maximum	48.5
44	123	NRU	TX	SLS-110	SLS-110	4	TRTDU	Treatment Duration Unit	HOURS
45	123	NRU	TX	SLS-110	SLS-110	5	INCBTMP	Incubation Temperature	37
46	123	NRU	TX	SLS-110	SLS-110	6	INCBTMPU	Incubation Temperature Unit	C
47	123	NRU	TX	SLS-110	SLS-110	7	WAVELGTH	Spectrophotometer wavelength in nanometers	540
48	123	NRU	TX	SLS-110	SLS-110	8	HUMID	Atmospheric Relative Humidity Percent	90
49	123	NRU	TX	SLS-110	SLS-110	9	ATMCO2	Atmospheric CO2 Percent	5
50	123	NRU	TX	SLS-110	SLS-110	10	SPTOBID	Sponsor defined tobacco identifier	CIG01a
51	123	NRU	TX	SLS-110	SLS-110	11	EXPTYP	Exposure Type	Air Liquid Interface
52	123	NRU	TX	SLS-110	SLS-110	12	SAMTYP	Sample Type	Liquid
53	123	NRU	TX	SLS-110	SLS-110	13	INTRVN	Name of the Intervention Article	Sodium Laurel Sulfate
54	123	NRU	TX	SLS-110	SLS-110	14	ITVTYPE	type of intervention article	POSITIVE CONTROL
55	123	NRU	TX	SLS-110	SLS-110	15	ITVCONC	intervention concentration	200
56	123	NRU	TX	SLS-110	SLS-110	16	ITVCONCU	intervention unit	ug/ml
57	123	NRU	TX	SLS-110	SLS-110	17	STRAIN	Strain/Substrain	CHO
58	123	NRU	TX	SLS-110	SLS-110	18	SPDEVID	Sponsor defined device identifier	PUFFMASTER3k
59	123	NRU	TX	SLS-110	SLS-110	19	DUREFID	Smoking Regime	Medium Intensity Regimen
60	123	NRU	TX	SLS-110	SLS-110	20	SPTOBID	Sponsor defined tobacco identifier	CIG01a

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Rows 1-2:	Show two records for TSPARMCD = "GLPTYP", using TSSEQ to indicate multiple records, since both GLP Types apply for this example study.
Row 1:	Shows the REFID of 123NRU that indicates the entire assay.
Rows 2, 5, 8, 9:	Show the REFIDs at the second level, all within the parent assay of 123NRU (REFID values of 1-3b, 1-3, 1-10, 2-10).
Rows 3-4, 6-7, 10-11:	Show REFID values that the sponsor chose, in this case, to represent the plate number (1 or 2), column (1 - 10), and row (A-G) of the 96-well plate.
Rows 3-4:	Show the REFIDs 1-3-A and 1-3-H that indicate two experimental units within the parent Trial Set of 1-3b.
Rows 6-7:	Show the REFIDs 1-3-B and 1-3-C that indicate two experimental units within the parent Trial Set of 1-3b.
Rows 10-11:	Show the REFIDs 1-10-G and 2-10-G that indicate two experimental units within the parent Trial Set of 1-3b.

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Row	STUDYID	ASSAYID	SETCD	REFTYPE	REFID	PARENT	LEVEL
1	123	NRU	ALL	ASSAY	123NRU		1
2	123	NRU	CSC-50b	TRIAL SET	1-3b	123NRU	2
3	123	NRU	CSC-50b	EXPERIMENTAL UNIT	1-3-A	1-3b	3
4	123	NRU	CSC-50b	EXPERIMENTAL UNIT	1-3-H	1-3b	3
5	123	NRU	CSC-50	TRIAL SET	1-3	123NRU	2
6	123	NRU	CSC-50	EXPERIMENTAL UNIT	1-3-B	1-3	3
7	123	NRU	CSC-50	EXPERIMENTAL UNIT	1-3-C	1-3	3
8	123	NRU	SLS-110	TRIAL SET	1-10	123NRU	2
9	123	NRU	SLS-110	TRIAL SET	2-10	123NRU	2
10	123	NRU	SLS-110	EXPERIMENTAL UNIT	1-10-G	1-10	3
11	123	NRU	SLS-110	EXPERIMENTAL UNIT	2-10-G	2-10	3

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cells growing in plate, treat cells in the plate, then add NRU dye, then extract NRU (the OBUID) dye and transfer to another plate (put in spectrophotometer & read).

row 161: This shows the EC50 result value for the entire assay (the SETCD, EUID, and OBUID are null).

Rows 1, 4: These data are not shown in the report table but included here for completeness of the NRU assay.

Row	STUDYID	ASSAYID	DOMAIN	GTSEQ	REFID	GTTESTCD	GTTEST	GTORRES	GTORRESU	GTCOLSRT (coll. summ. result type)	GTSTRESC	GTSTRESN	GTSTRESU
1	123	NRU	GT	1	1-3-B	ABSORB	Absorbance Reading	1.7 (0.0 - 2.0)			1.7	1.7
2	123	NRU	GT	2	1-3-B	BKCABS	Background Corrected Absorbance Reading	1.6			1.6	1.6
3	123	NRU	GT	3	1-3-B	RELABS	Relative Absorbance Reading	77.7	%		77.7	77.7	%
4	123	NRU	GT	4	1-3-C	ABSORB	Absorbance Reading	1.8 (0.0 - 2.0)			1.8	1.8
5	123	NRU	GT	5	1-3-C	BKCABS	Background Corrected Absorbance Reading	1.7			1.7	1.7
6	123	NRU	GT	6	1-3-C	RELABS	Relative Absorbance Reading	77.4			77.4	77.4
7	123	NRU	GT	7	1-3	RELABS	Relative Absorbance Reading	78.2	%	MEAN	78.2	78.2	%
8	123	NRU	GT	8	1-3	RELABS	Relative Absorbance Reading	2.9	%	STANDARD DEVIATION	2.9	2.9	%
9	123	NRU	GT	9	1-3	RELABS	Relative Absorbance Reading	3.6		COEFFICIENT OF VARIANCE	3.6	3.6
10	123	NRU	GT	10	1-10-G	ABSORB	Absorbance Reading	0.7			0.7	0.7
11	123	NRU	GT	11	1-10-G	BKCABS	Background Corrected Absorbance Reading	0.6			0.6	0.6
12	123	NRU	GT	11	1-10-G	RELABS	Relative Absorbance Reading	8.57			8.57	8.57
13	123	NRU	GT	12	1-10	RELABS	Relative Absorbance Reading	8.94	%	MEAN	8.94	8.94
14	123	NRU	GT	13	1-10	RELABS	Relative Absorbance Reading	2.72	%	STANDARD DEVIATION	2.72	2.72
15	123	NRU	GT	14	1-10	RELABS	Relative Absorbance Reading	30.5		COEFFICIENT OF VARIANCE	30.5	30.5
16	123	NRU	GT	15	2-10-G	ABSORB	Absorbance Reading	0.9
17	123	NRU	GT	16	2-10-G	BKCABS	Background Corrected Absorbance Reading	0.8
18	123	NRU	GT	17	2-10-G	RELABS	Relative Absorbance Reading	6.30			6.30	6.30
19	123	NRU	GT	18	2-10	RELABS	Relative Absorbance Reading	5.91	%	MEAN	5.91	5.91
20	123	NRU	GT	19	2-10	RELABS	Relative Absorbance Reading	0.83	%	STANDARD DEVIATION	0.83	0.83
21	123	NRU	GT	20	2-10	RELABS	Relative Absorbance Reading	14.1		COEFFICIENT OF VARIANCE	14.1	14.1
22	123	NRU	GT	21	123NRU	EC50	Effective Concentration of 50 Percent Cytotoxicity	123	ug/ml		123	123	ug/ml

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SEND Example.In vitro Neutral Red Cytotoxicity Assay

Row

STUDYID

ASSAYID

DOMAIN

SETCD

SET

TXSEQ

TXPARMCD

TXPARM

TXVAL